                         SEQUENCE LISTING

<110>  GlaxoSmithKline Biologicals s.a.
 
<120>  METHODS FOR PROTEIN PURIFICATION

<130>  VB66773 WO

<150>  EP19183033.0
<151>  2019-06-27

<160>  47    

<170>  PatentIn version 3.5

<210>  1
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of H4 tag

<400>  1

His His His His 
1               


<210>  2
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of R4 tag

<400>  2

Arg Arg Arg Arg 
1               


<210>  3
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of H2R2 tag

<400>  3

His His Arg Arg 
1               


<210>  4
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of (HR)2 tag

<400>  4

His Arg His Arg 
1               


<210>  5
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of (HR)3 tag

<400>  5

His Arg His Arg His Arg 
1               5       


<210>  6
<211>  8
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of (HR)4 tag

<400>  6

His Arg His Arg His Arg His Arg 
1               5               


<210>  7
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of R6 tag

<400>  7

Arg Arg Arg Arg Arg Arg 
1               5       


<210>  8
<211>  12
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of (PR)6 tag

<400>  8

Pro Arg Pro Arg Pro Arg Pro Arg Pro Arg Pro Arg 
1               5                   10          


<210>  9
<211>  12
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of (PSR)4 tag

<400>  9

Pro Ser Arg Pro Ser Arg Pro Ser Arg Pro Ser Arg 
1               5                   10          


<210>  10
<211>  613
<212>  PRT
<213>  Pseudomonas aeruginosa

<400>  10

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Tyr 
        195                 200                 205             


Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp Thr Trp Glu Gly Lys Ile 
    210                 215                 220                 


Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys His Asp Leu Asp Ile Lys 
225                 230                 235                 240 


Pro Thr Val Ile Ser His Arg Leu His Phe Pro Glu Gly Gly Ser Leu 
                245                 250                 255     


Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Ala Phe 
            260                 265                 270         


Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly 
        275                 280                 285             


Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser 
    290                 295                 300                 


Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly 
305                 310                 315                 320 


Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala 
                325                 330                 335     


Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg 
            340                 345                 350         


Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val 
        355                 360                 365             


Ser Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp 
    370                 375                 380                 


Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe 
385                 390                 395                 400 


Leu Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn 
                405                 410                 415     


Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg 
            420                 425                 430         


Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln 
        435                 440                 445             


Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala 
    450                 455                 460                 


Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly 
465                 470                 475                 480 


Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly 
                485                 490                 495     


Ala Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr 
            500                 505                 510         


Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu 
        515                 520                 525             


Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly 
    530                 535                 540                 


Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro Leu 
545                 550                 555                 560 


Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg 
                565                 570                 575     


Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln 
            580                 585                 590         


Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro 
        595                 600                 605             


Arg Glu Asp Leu Lys 
    610             


<210>  11
<211>  612
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of EPA with L552V/?E553 detoxifying mutation 
       (bold, underlined)

<400>  11

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Tyr 
        195                 200                 205             


Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp Thr Trp Glu Gly Lys Ile 
    210                 215                 220                 


Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys His Asp Leu Asp Ile Lys 
225                 230                 235                 240 


Pro Thr Val Ile Ser His Arg Leu His Phe Pro Glu Gly Gly Ser Leu 
                245                 250                 255     


Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Ala Phe 
            260                 265                 270         


Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly 
        275                 280                 285             


Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser 
    290                 295                 300                 


Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly 
305                 310                 315                 320 


Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala 
                325                 330                 335     


Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg 
            340                 345                 350         


Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val 
        355                 360                 365             


Ser Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp 
    370                 375                 380                 


Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe 
385                 390                 395                 400 


Leu Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn 
                405                 410                 415     


Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg 
            420                 425                 430         


Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln 
        435                 440                 445             


Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala 
    450                 455                 460                 


Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly 
465                 470                 475                 480 


Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly 
                485                 490                 495     


Ala Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr 
            500                 505                 510         


Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu 
        515                 520                 525             


Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly 
    530                 535                 540                 


Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu Gly Trp Pro Leu Ala 
545                 550                 555                 560 


Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn 
                565                 570                 575     


Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala 
            580                 585                 590         


Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg 
        595                 600                 605             


Glu Asp Leu Lys 
    610         


<210>  12
<211>  616
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation and HR2 tag

<400>  12

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Tyr 
        195                 200                 205             


Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp Thr Trp Glu Gly Lys Ile 
    210                 215                 220                 


Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys His Asp Leu Asp Ile Lys 
225                 230                 235                 240 


Pro Thr Val Ile Ser His Arg Leu His Phe Pro Glu Gly Gly Ser Leu 
                245                 250                 255     


Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Ala Phe 
            260                 265                 270         


Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly 
        275                 280                 285             


Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser 
    290                 295                 300                 


Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly 
305                 310                 315                 320 


Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala 
                325                 330                 335     


Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg 
            340                 345                 350         


Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val 
        355                 360                 365             


Ser Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp 
    370                 375                 380                 


Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe 
385                 390                 395                 400 


Leu Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn 
                405                 410                 415     


Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg 
            420                 425                 430         


Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln 
        435                 440                 445             


Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala 
    450                 455                 460                 


Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly 
465                 470                 475                 480 


Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly 
                485                 490                 495     


Ala Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr 
            500                 505                 510         


Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu 
        515                 520                 525             


Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly 
    530                 535                 540                 


Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu Gly Trp Pro Leu Ala 
545                 550                 555                 560 


Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn 
                565                 570                 575     


Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala 
            580                 585                 590         


Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg 
        595                 600                 605             


Glu Asp Leu Lys His Arg His Arg 
    610                 615     


<210>  13
<211>  618
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation and HR3 tag

<400>  13

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Tyr 
        195                 200                 205             


Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp Thr Trp Glu Gly Lys Ile 
    210                 215                 220                 


Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys His Asp Leu Asp Ile Lys 
225                 230                 235                 240 


Pro Thr Val Ile Ser His Arg Leu His Phe Pro Glu Gly Gly Ser Leu 
                245                 250                 255     


Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Ala Phe 
            260                 265                 270         


Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly 
        275                 280                 285             


Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser 
    290                 295                 300                 


Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly 
305                 310                 315                 320 


Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala 
                325                 330                 335     


Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg 
            340                 345                 350         


Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val 
        355                 360                 365             


Ser Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp 
    370                 375                 380                 


Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe 
385                 390                 395                 400 


Leu Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn 
                405                 410                 415     


Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg 
            420                 425                 430         


Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln 
        435                 440                 445             


Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala 
    450                 455                 460                 


Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly 
465                 470                 475                 480 


Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly 
                485                 490                 495     


Ala Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr 
            500                 505                 510         


Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu 
        515                 520                 525             


Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly 
    530                 535                 540                 


Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu Gly Trp Pro Leu Ala 
545                 550                 555                 560 


Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn 
                565                 570                 575     


Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala 
            580                 585                 590         


Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg 
        595                 600                 605             


Glu Asp Leu Lys His Arg His Arg His Arg 
    610                 615             


<210>  14
<211>  620
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation and HR4 tag

<400>  14

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Tyr 
        195                 200                 205             


Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp Thr Trp Glu Gly Lys Ile 
    210                 215                 220                 


Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys His Asp Leu Asp Ile Lys 
225                 230                 235                 240 


Pro Thr Val Ile Ser His Arg Leu His Phe Pro Glu Gly Gly Ser Leu 
                245                 250                 255     


Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Ala Phe 
            260                 265                 270         


Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly 
        275                 280                 285             


Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser 
    290                 295                 300                 


Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly 
305                 310                 315                 320 


Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala 
                325                 330                 335     


Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg 
            340                 345                 350         


Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val 
        355                 360                 365             


Ser Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp 
    370                 375                 380                 


Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe 
385                 390                 395                 400 


Leu Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn 
                405                 410                 415     


Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg 
            420                 425                 430         


Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln 
        435                 440                 445             


Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala 
    450                 455                 460                 


Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly 
465                 470                 475                 480 


Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly 
                485                 490                 495     


Ala Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr 
            500                 505                 510         


Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu 
        515                 520                 525             


Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly 
    530                 535                 540                 


Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu Gly Trp Pro Leu Ala 
545                 550                 555                 560 


Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn 
                565                 570                 575     


Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala 
            580                 585                 590         


Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg 
        595                 600                 605             


Glu Asp Leu Lys His Arg His Arg His Arg His Arg 
    610                 615                 620 


<210>  15
<211>  616
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation and R4 tag

<400>  15

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Tyr 
        195                 200                 205             


Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp Thr Trp Glu Gly Lys Ile 
    210                 215                 220                 


Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys His Asp Leu Asp Ile Lys 
225                 230                 235                 240 


Pro Thr Val Ile Ser His Arg Leu His Phe Pro Glu Gly Gly Ser Leu 
                245                 250                 255     


Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Ala Phe 
            260                 265                 270         


Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly 
        275                 280                 285             


Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser 
    290                 295                 300                 


Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly 
305                 310                 315                 320 


Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala 
                325                 330                 335     


Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg 
            340                 345                 350         


Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val 
        355                 360                 365             


Ser Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp 
    370                 375                 380                 


Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe 
385                 390                 395                 400 


Leu Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn 
                405                 410                 415     


Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg 
            420                 425                 430         


Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln 
        435                 440                 445             


Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala 
    450                 455                 460                 


Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly 
465                 470                 475                 480 


Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly 
                485                 490                 495     


Ala Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr 
            500                 505                 510         


Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu 
        515                 520                 525             


Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly 
    530                 535                 540                 


Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu Gly Trp Pro Leu Ala 
545                 550                 555                 560 


Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn 
                565                 570                 575     


Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala 
            580                 585                 590         


Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg 
        595                 600                 605             


Glu Asp Leu Lys Arg Arg Arg Arg 
    610                 615     


<210>  16
<211>  618
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation and R6 tag

<400>  16

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Tyr 
        195                 200                 205             


Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp Thr Trp Glu Gly Lys Ile 
    210                 215                 220                 


Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys His Asp Leu Asp Ile Lys 
225                 230                 235                 240 


Pro Thr Val Ile Ser His Arg Leu His Phe Pro Glu Gly Gly Ser Leu 
                245                 250                 255     


Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Ala Phe 
            260                 265                 270         


Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly 
        275                 280                 285             


Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser 
    290                 295                 300                 


Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly 
305                 310                 315                 320 


Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala 
                325                 330                 335     


Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg 
            340                 345                 350         


Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val 
        355                 360                 365             


Ser Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp 
    370                 375                 380                 


Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe 
385                 390                 395                 400 


Leu Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn 
                405                 410                 415     


Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg 
            420                 425                 430         


Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln 
        435                 440                 445             


Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala 
    450                 455                 460                 


Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly 
465                 470                 475                 480 


Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly 
                485                 490                 495     


Ala Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr 
            500                 505                 510         


Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu 
        515                 520                 525             


Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly 
    530                 535                 540                 


Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu Gly Trp Pro Leu Ala 
545                 550                 555                 560 


Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn 
                565                 570                 575     


Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala 
            580                 585                 590         


Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg 
        595                 600                 605             


Glu Asp Leu Lys Arg Arg Arg Arg Arg Arg 
    610                 615             


<210>  17
<211>  624
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation and PR6 tag

<400>  17

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Tyr 
        195                 200                 205             


Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp Thr Trp Glu Gly Lys Ile 
    210                 215                 220                 


Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys His Asp Leu Asp Ile Lys 
225                 230                 235                 240 


Pro Thr Val Ile Ser His Arg Leu His Phe Pro Glu Gly Gly Ser Leu 
                245                 250                 255     


Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Ala Phe 
            260                 265                 270         


Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly 
        275                 280                 285             


Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser 
    290                 295                 300                 


Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly 
305                 310                 315                 320 


Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala 
                325                 330                 335     


Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg 
            340                 345                 350         


Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val 
        355                 360                 365             


Ser Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp 
    370                 375                 380                 


Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe 
385                 390                 395                 400 


Leu Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn 
                405                 410                 415     


Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg 
            420                 425                 430         


Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln 
        435                 440                 445             


Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala 
    450                 455                 460                 


Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly 
465                 470                 475                 480 


Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly 
                485                 490                 495     


Ala Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr 
            500                 505                 510         


Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu 
        515                 520                 525             


Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly 
    530                 535                 540                 


Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu Gly Trp Pro Leu Ala 
545                 550                 555                 560 


Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn 
                565                 570                 575     


Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala 
            580                 585                 590         


Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg 
        595                 600                 605             


Glu Asp Leu Lys Pro Arg Pro Arg Pro Arg Pro Arg Pro Arg Pro Arg 
    610                 615                 620                 


<210>  18
<211>  624
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation and PSR4 tag

<400>  18

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Tyr 
        195                 200                 205             


Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp Thr Trp Glu Gly Lys Ile 
    210                 215                 220                 


Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys His Asp Leu Asp Ile Lys 
225                 230                 235                 240 


Pro Thr Val Ile Ser His Arg Leu His Phe Pro Glu Gly Gly Ser Leu 
                245                 250                 255     


Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Ala Phe 
            260                 265                 270         


Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly 
        275                 280                 285             


Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser 
    290                 295                 300                 


Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly 
305                 310                 315                 320 


Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala 
                325                 330                 335     


Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg 
            340                 345                 350         


Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val 
        355                 360                 365             


Ser Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp 
    370                 375                 380                 


Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe 
385                 390                 395                 400 


Leu Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn 
                405                 410                 415     


Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg 
            420                 425                 430         


Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln 
        435                 440                 445             


Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala 
    450                 455                 460                 


Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly 
465                 470                 475                 480 


Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly 
                485                 490                 495     


Ala Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr 
            500                 505                 510         


Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu 
        515                 520                 525             


Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly 
    530                 535                 540                 


Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu Gly Trp Pro Leu Ala 
545                 550                 555                 560 


Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn 
                565                 570                 575     


Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala 
            580                 585                 590         


Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg 
        595                 600                 605             


Glu Asp Leu Lys Pro Ser Arg Pro Ser Arg Pro Ser Arg Pro Ser Arg 
    610                 615                 620                 


<210>  19
<211>  293
<212>  PRT
<213>  Staphylococcus aureus

<400>  19

Ala Asp Ser Asp Ile Asn Ile Lys Thr Gly Thr Thr Asp Ile Gly Ser 
1               5                   10                  15      


Asn Thr Thr Val Lys Thr Gly Asp Leu Val Thr Tyr Asp Lys Glu Asn 
            20                  25                  30          


Gly Met His Lys Lys Val Phe Tyr Ser Phe Ile Asp Asp Lys Asn His 
        35                  40                  45              


Asn Lys Lys Leu Leu Val Ile Arg Thr Lys Gly Thr Ile Ala Gly Gln 
    50                  55                  60                  


Tyr Arg Val Tyr Ser Glu Glu Gly Ala Asn Lys Ser Gly Leu Ala Trp 
65                  70                  75                  80  


Pro Ser Ala Phe Lys Val Gln Leu Gln Leu Pro Asp Asn Glu Val Ala 
                85                  90                  95      


Gln Ile Ser Asp Tyr Tyr Pro Arg Asn Ser Ile Asp Thr Lys Glu Tyr 
            100                 105                 110         


Met Ser Thr Leu Thr Tyr Gly Phe Asn Gly Asn Val Thr Gly Asp Asp 
        115                 120                 125             


Thr Gly Lys Ile Gly Gly Leu Ile Gly Ala Asn Val Ser Ile Gly His 
    130                 135                 140                 


Thr Leu Lys Tyr Val Gln Pro Asp Phe Lys Thr Ile Leu Glu Ser Pro 
145                 150                 155                 160 


Thr Asp Lys Lys Val Gly Trp Lys Val Ile Phe Asn Asn Met Val Asn 
                165                 170                 175     


Gln Asn Trp Gly Pro Tyr Asp Arg Asp Ser Trp Asn Pro Val Tyr Gly 
            180                 185                 190         


Asn Gln Leu Phe Met Lys Thr Arg Asn Gly Ser Met Lys Ala Ala Asp 
        195                 200                 205             


Asn Phe Leu Asp Pro Asn Lys Ala Ser Ser Leu Leu Ser Ser Gly Phe 
    210                 215                 220                 


Ser Pro Asp Phe Ala Thr Val Ile Thr Met Asp Arg Lys Ala Ser Lys 
225                 230                 235                 240 


Gln Gln Thr Asn Ile Asp Val Ile Tyr Glu Arg Val Arg Asp Asp Tyr 
                245                 250                 255     


Gln Leu His Trp Thr Ser Thr Asn Trp Lys Gly Thr Asn Thr Lys Asp 
            260                 265                 270         


Lys Trp Ile Asp Arg Ser Ser Glu Arg Tyr Lys Ile Asp Trp Glu Lys 
        275                 280                 285             


Glu Glu Met Thr Asn 
    290             


<210>  20
<211>  299
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of Hla with glycosite KDQNRTK substitutued 
       for K131, H35L detoxifying mutation, H48C/G122C stabilizing 
       mutations (bold, underlined)

<400>  20

Ala Asp Ser Asp Ile Asn Ile Lys Thr Gly Thr Thr Asp Ile Gly Ser 
1               5                   10                  15      


Asn Thr Thr Val Lys Thr Gly Asp Leu Val Thr Tyr Asp Lys Glu Asn 
            20                  25                  30          


Gly Met Leu Lys Lys Val Phe Tyr Ser Phe Ile Asp Asp Lys Asn Cys 
        35                  40                  45              


Asn Lys Lys Leu Leu Val Ile Arg Thr Lys Gly Thr Ile Ala Gly Gln 
    50                  55                  60                  


Tyr Arg Val Tyr Ser Glu Glu Gly Ala Asn Lys Ser Gly Leu Ala Trp 
65                  70                  75                  80  


Pro Ser Ala Phe Lys Val Gln Leu Gln Leu Pro Asp Asn Glu Val Ala 
                85                  90                  95      


Gln Ile Ser Asp Tyr Tyr Pro Arg Asn Ser Ile Asp Thr Lys Glu Tyr 
            100                 105                 110         


Met Ser Thr Leu Thr Tyr Gly Phe Asn Cys Asn Val Thr Gly Asp Asp 
        115                 120                 125             


Thr Gly Lys Asp Gln Asn Arg Thr Lys Ile Gly Gly Leu Ile Gly Ala 
    130                 135                 140                 


Asn Val Ser Ile Gly His Thr Leu Lys Tyr Val Gln Pro Asp Phe Lys 
145                 150                 155                 160 


Thr Ile Leu Glu Ser Pro Thr Asp Lys Lys Val Gly Trp Lys Val Ile 
                165                 170                 175     


Phe Asn Asn Met Val Asn Gln Asn Trp Gly Pro Tyr Asp Arg Asp Ser 
            180                 185                 190         


Trp Asn Pro Val Tyr Gly Asn Gln Leu Phe Met Lys Thr Arg Asn Gly 
        195                 200                 205             


Ser Met Lys Ala Ala Asp Asn Phe Leu Asp Pro Asn Lys Ala Ser Ser 
    210                 215                 220                 


Leu Leu Ser Ser Gly Phe Ser Pro Asp Phe Ala Thr Val Ile Thr Met 
225                 230                 235                 240 


Asp Arg Lys Ala Ser Lys Gln Gln Thr Asn Ile Asp Val Ile Tyr Glu 
                245                 250                 255     


Arg Val Arg Asp Asp Tyr Gln Leu His Trp Thr Ser Thr Asn Trp Lys 
            260                 265                 270         


Gly Thr Asn Thr Lys Asp Lys Trp Ile Asp Arg Ser Ser Glu Arg Tyr 
        275                 280                 285             


Lys Ile Asp Trp Glu Lys Glu Glu Met Thr Asn 
    290                 295                 


<210>  21
<211>  305
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of Hla with glycosite, detoxifying and 
       stabilizing mutations, linker and H4 tag

<400>  21

Ala Asp Ser Asp Ile Asn Ile Lys Thr Gly Thr Thr Asp Ile Gly Ser 
1               5                   10                  15      


Asn Thr Thr Val Lys Thr Gly Asp Leu Val Thr Tyr Asp Lys Glu Asn 
            20                  25                  30          


Gly Met Leu Lys Lys Val Phe Tyr Ser Phe Ile Asp Asp Lys Asn Cys 
        35                  40                  45              


Asn Lys Lys Leu Leu Val Ile Arg Thr Lys Gly Thr Ile Ala Gly Gln 
    50                  55                  60                  


Tyr Arg Val Tyr Ser Glu Glu Gly Ala Asn Lys Ser Gly Leu Ala Trp 
65                  70                  75                  80  


Pro Ser Ala Phe Lys Val Gln Leu Gln Leu Pro Asp Asn Glu Val Ala 
                85                  90                  95      


Gln Ile Ser Asp Tyr Tyr Pro Arg Asn Ser Ile Asp Thr Lys Glu Tyr 
            100                 105                 110         


Met Ser Thr Leu Thr Tyr Gly Phe Asn Cys Asn Val Thr Gly Asp Asp 
        115                 120                 125             


Thr Gly Lys Asp Gln Asn Arg Thr Lys Ile Gly Gly Leu Ile Gly Ala 
    130                 135                 140                 


Asn Val Ser Ile Gly His Thr Leu Lys Tyr Val Gln Pro Asp Phe Lys 
145                 150                 155                 160 


Thr Ile Leu Glu Ser Pro Thr Asp Lys Lys Val Gly Trp Lys Val Ile 
                165                 170                 175     


Phe Asn Asn Met Val Asn Gln Asn Trp Gly Pro Tyr Asp Arg Asp Ser 
            180                 185                 190         


Trp Asn Pro Val Tyr Gly Asn Gln Leu Phe Met Lys Thr Arg Asn Gly 
        195                 200                 205             


Ser Met Lys Ala Ala Asp Asn Phe Leu Asp Pro Asn Lys Ala Ser Ser 
    210                 215                 220                 


Leu Leu Ser Ser Gly Phe Ser Pro Asp Phe Ala Thr Val Ile Thr Met 
225                 230                 235                 240 


Asp Arg Lys Ala Ser Lys Gln Gln Thr Asn Ile Asp Val Ile Tyr Glu 
                245                 250                 255     


Arg Val Arg Asp Asp Tyr Gln Leu His Trp Thr Ser Thr Asn Trp Lys 
            260                 265                 270         


Gly Thr Asn Thr Lys Asp Lys Trp Ile Asp Arg Ser Ser Glu Arg Tyr 
        275                 280                 285             


Lys Ile Asp Trp Glu Lys Glu Glu Met Thr Asn Gly Ser His His His 
    290                 295                 300                 


His 
305 


<210>  22
<211>  305
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of Hla with glycosite, detoxifying and 
       stabilizing mutations, linker and R4 tag

<400>  22

Ala Asp Ser Asp Ile Asn Ile Lys Thr Gly Thr Thr Asp Ile Gly Ser 
1               5                   10                  15      


Asn Thr Thr Val Lys Thr Gly Asp Leu Val Thr Tyr Asp Lys Glu Asn 
            20                  25                  30          


Gly Met Leu Lys Lys Val Phe Tyr Ser Phe Ile Asp Asp Lys Asn Cys 
        35                  40                  45              


Asn Lys Lys Leu Leu Val Ile Arg Thr Lys Gly Thr Ile Ala Gly Gln 
    50                  55                  60                  


Tyr Arg Val Tyr Ser Glu Glu Gly Ala Asn Lys Ser Gly Leu Ala Trp 
65                  70                  75                  80  


Pro Ser Ala Phe Lys Val Gln Leu Gln Leu Pro Asp Asn Glu Val Ala 
                85                  90                  95      


Gln Ile Ser Asp Tyr Tyr Pro Arg Asn Ser Ile Asp Thr Lys Glu Tyr 
            100                 105                 110         


Met Ser Thr Leu Thr Tyr Gly Phe Asn Cys Asn Val Thr Gly Asp Asp 
        115                 120                 125             


Thr Gly Lys Asp Gln Asn Arg Thr Lys Ile Gly Gly Leu Ile Gly Ala 
    130                 135                 140                 


Asn Val Ser Ile Gly His Thr Leu Lys Tyr Val Gln Pro Asp Phe Lys 
145                 150                 155                 160 


Thr Ile Leu Glu Ser Pro Thr Asp Lys Lys Val Gly Trp Lys Val Ile 
                165                 170                 175     


Phe Asn Asn Met Val Asn Gln Asn Trp Gly Pro Tyr Asp Arg Asp Ser 
            180                 185                 190         


Trp Asn Pro Val Tyr Gly Asn Gln Leu Phe Met Lys Thr Arg Asn Gly 
        195                 200                 205             


Ser Met Lys Ala Ala Asp Asn Phe Leu Asp Pro Asn Lys Ala Ser Ser 
    210                 215                 220                 


Leu Leu Ser Ser Gly Phe Ser Pro Asp Phe Ala Thr Val Ile Thr Met 
225                 230                 235                 240 


Asp Arg Lys Ala Ser Lys Gln Gln Thr Asn Ile Asp Val Ile Tyr Glu 
                245                 250                 255     


Arg Val Arg Asp Asp Tyr Gln Leu His Trp Thr Ser Thr Asn Trp Lys 
            260                 265                 270         


Gly Thr Asn Thr Lys Asp Lys Trp Ile Asp Arg Ser Ser Glu Arg Tyr 
        275                 280                 285             


Lys Ile Asp Trp Glu Lys Glu Glu Met Thr Asn Gly Ser Arg Arg Arg 
    290                 295                 300                 


Arg 
305 


<210>  23
<211>  305
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of Hla with glycosite, detoxifying and 
       stabilizing mutations, linker and H2R2 tag

<400>  23

Ala Asp Ser Asp Ile Asn Ile Lys Thr Gly Thr Thr Asp Ile Gly Ser 
1               5                   10                  15      


Asn Thr Thr Val Lys Thr Gly Asp Leu Val Thr Tyr Asp Lys Glu Asn 
            20                  25                  30          


Gly Met Leu Lys Lys Val Phe Tyr Ser Phe Ile Asp Asp Lys Asn Cys 
        35                  40                  45              


Asn Lys Lys Leu Leu Val Ile Arg Thr Lys Gly Thr Ile Ala Gly Gln 
    50                  55                  60                  


Tyr Arg Val Tyr Ser Glu Glu Gly Ala Asn Lys Ser Gly Leu Ala Trp 
65                  70                  75                  80  


Pro Ser Ala Phe Lys Val Gln Leu Gln Leu Pro Asp Asn Glu Val Ala 
                85                  90                  95      


Gln Ile Ser Asp Tyr Tyr Pro Arg Asn Ser Ile Asp Thr Lys Glu Tyr 
            100                 105                 110         


Met Ser Thr Leu Thr Tyr Gly Phe Asn Cys Asn Val Thr Gly Asp Asp 
        115                 120                 125             


Thr Gly Lys Asp Gln Asn Arg Thr Lys Ile Gly Gly Leu Ile Gly Ala 
    130                 135                 140                 


Asn Val Ser Ile Gly His Thr Leu Lys Tyr Val Gln Pro Asp Phe Lys 
145                 150                 155                 160 


Thr Ile Leu Glu Ser Pro Thr Asp Lys Lys Val Gly Trp Lys Val Ile 
                165                 170                 175     


Phe Asn Asn Met Val Asn Gln Asn Trp Gly Pro Tyr Asp Arg Asp Ser 
            180                 185                 190         


Trp Asn Pro Val Tyr Gly Asn Gln Leu Phe Met Lys Thr Arg Asn Gly 
        195                 200                 205             


Ser Met Lys Ala Ala Asp Asn Phe Leu Asp Pro Asn Lys Ala Ser Ser 
    210                 215                 220                 


Leu Leu Ser Ser Gly Phe Ser Pro Asp Phe Ala Thr Val Ile Thr Met 
225                 230                 235                 240 


Asp Arg Lys Ala Ser Lys Gln Gln Thr Asn Ile Asp Val Ile Tyr Glu 
                245                 250                 255     


Arg Val Arg Asp Asp Tyr Gln Leu His Trp Thr Ser Thr Asn Trp Lys 
            260                 265                 270         


Gly Thr Asn Thr Lys Asp Lys Trp Ile Asp Arg Ser Ser Glu Arg Tyr 
        275                 280                 285             


Lys Ile Asp Trp Glu Lys Glu Glu Met Thr Asn Gly Ser His His Arg 
    290                 295                 300                 


Arg 
305 


<210>  24
<211>  305
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of Hla with glycosite, detoxifying and 
       stabilizing mutations, linker and (HR)2 tag

<400>  24

Ala Asp Ser Asp Ile Asn Ile Lys Thr Gly Thr Thr Asp Ile Gly Ser 
1               5                   10                  15      


Asn Thr Thr Val Lys Thr Gly Asp Leu Val Thr Tyr Asp Lys Glu Asn 
            20                  25                  30          


Gly Met Leu Lys Lys Val Phe Tyr Ser Phe Ile Asp Asp Lys Asn Cys 
        35                  40                  45              


Asn Lys Lys Leu Leu Val Ile Arg Thr Lys Gly Thr Ile Ala Gly Gln 
    50                  55                  60                  


Tyr Arg Val Tyr Ser Glu Glu Gly Ala Asn Lys Ser Gly Leu Ala Trp 
65                  70                  75                  80  


Pro Ser Ala Phe Lys Val Gln Leu Gln Leu Pro Asp Asn Glu Val Ala 
                85                  90                  95      


Gln Ile Ser Asp Tyr Tyr Pro Arg Asn Ser Ile Asp Thr Lys Glu Tyr 
            100                 105                 110         


Met Ser Thr Leu Thr Tyr Gly Phe Asn Cys Asn Val Thr Gly Asp Asp 
        115                 120                 125             


Thr Gly Lys Asp Gln Asn Arg Thr Lys Ile Gly Gly Leu Ile Gly Ala 
    130                 135                 140                 


Asn Val Ser Ile Gly His Thr Leu Lys Tyr Val Gln Pro Asp Phe Lys 
145                 150                 155                 160 


Thr Ile Leu Glu Ser Pro Thr Asp Lys Lys Val Gly Trp Lys Val Ile 
                165                 170                 175     


Phe Asn Asn Met Val Asn Gln Asn Trp Gly Pro Tyr Asp Arg Asp Ser 
            180                 185                 190         


Trp Asn Pro Val Tyr Gly Asn Gln Leu Phe Met Lys Thr Arg Asn Gly 
        195                 200                 205             


Ser Met Lys Ala Ala Asp Asn Phe Leu Asp Pro Asn Lys Ala Ser Ser 
    210                 215                 220                 


Leu Leu Ser Ser Gly Phe Ser Pro Asp Phe Ala Thr Val Ile Thr Met 
225                 230                 235                 240 


Asp Arg Lys Ala Ser Lys Gln Gln Thr Asn Ile Asp Val Ile Tyr Glu 
                245                 250                 255     


Arg Val Arg Asp Asp Tyr Gln Leu His Trp Thr Ser Thr Asn Trp Lys 
            260                 265                 270         


Gly Thr Asn Thr Lys Asp Lys Trp Ile Asp Arg Ser Ser Glu Arg Tyr 
        275                 280                 285             


Lys Ile Asp Trp Glu Lys Glu Glu Met Thr Asn Gly Ser His Arg His 
    290                 295                 300                 


Arg 
305 


<210>  25
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Minimal PglB glycosite consensus sequence


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  Xaa can be Asp or Glu'

<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  Xaa can be any naturally occurring amino acid except proline

<220>
<221>  MISC_FEATURE
<222>  (4)..(4)
<223>  Xaa can be any naturally occurring amino acid except proline

<220>
<221>  MISC_FEATURE
<222>  (5)..(5)
<223>  Xaa can be Ser or Thr'

<220>
<221>  MISC_FEATURE
<222>  (5)..(5)
<223>  Xaa can be Ser or Thr

<400>  25

Xaa Xaa Asn Xaa Xaa 
1               5   


<210>  26
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Full PglB glycosite consensus sequence


<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  Xaa can be Asp or Glu

<220>
<221>  MISC_FEATURE
<222>  (3)..(3)
<223>  Xaa can be any naturally occurring amino acid except proline

<220>
<221>  MISC_FEATURE
<222>  (5)..(5)
<223>  Xaa can be any naturally occurring amino acid except proline

<220>
<221>  MISC_FEATURE
<222>  (6)..(6)
<223>  Xaa can be Ser or Thr

<400>  26

Lys Xaa Xaa Asn Xaa Xaa Lys 
1               5           


<210>  27
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  PglB glycosite sequence (Hla)

<400>  27

Lys Asp Gln Asn Arg Thr Lys 
1               5           


<210>  28
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  PglB glycosite sequence  (EPA)

<400>  28

Lys Asp Gln Asn Ala Thr Lys 
1               5           


<210>  29
<211>  19
<212>  PRT
<213>  Escherichia coli

<400>  29

Met Ile Lys Phe Leu Ser Ala Leu Ile Leu Leu Leu Val Thr Thr Ala 
1               5                   10                  15      


Ala Gln Ala 
            


<210>  30
<211>  21
<212>  PRT
<213>  Escherichia coli

<400>  30

Met Lys Lys Thr Ala Ile Ala Ile Ala Val Ala Leu Ala Gly Phe Ala 
1               5                   10                  15      


Thr Val Ala Gln Ala 
            20      


<210>  31
<211>  26
<212>  PRT
<213>  Escherichia coli

<400>  31

Met Lys Ile Lys Thr Gly Ala Arg Ile Leu Ala Leu Ser Ala Leu Thr 
1               5                   10                  15      


Thr Met Met Phe Ser Ala Ser Ala Leu Ala 
            20                  25      


<210>  32
<211>  22
<212>  PRT
<213>  Erwinia carotovora

<400>  32

Met Lys Tyr Leu Leu Pro Thr Ala Ala Ala Gly Leu Leu Leu Leu Ala 
1               5                   10                  15      


Ala Gln Pro Ala Met Ala 
            20          


<210>  33
<211>  23
<212>  PRT
<213>  Escherichia coli

<400>  33

Met Ser Phe Lys Lys Ile Ile Lys Ala Phe Val Ile Met Ala Ala Leu 
1               5                   10                  15      


Val Ser Val Gln Ala His Ala 
            20              


<210>  34
<211>  28
<212>  PRT
<213>  Escherichia coli

<400>  34

Met Phe Lys Phe Lys Lys Lys Phe Leu Val Gly Leu Thr Ala Ala Phe 
1               5                   10                  15      


Met Ser Ile Ser Met Phe Ser Ala Thr Ala Ser Ala 
            20                  25              


<210>  35
<211>  19
<212>  PRT
<213>  Escherichia coli

<400>  35

Met Lys Lys Ile Trp Leu Ala Leu Ala Gly Leu Val Leu Ala Phe Ser 
1               5                   10                  15      


Ala Ser Ala 
            


<210>  36
<211>  21
<212>  PRT
<213>  Escherichia coli

<400>  36

Met Lys Gln Ala Leu Arg Val Ala Phe Gly Phe Leu Ile Leu Trp Ala 
1               5                   10                  15      


Ser Val Leu His Ala 
            20      


<210>  37
<211>  26
<212>  PRT
<213>  Escherichia coli

<400>  37

Met Lys Met Asn Lys Lys Val Leu Leu Thr Ser Thr Met Ala Ala Ser 
1               5                   10                  15      


Leu Leu Ser Val Ala Ser Val Gln Ala Ser 
            20                  25      


<210>  38
<211>  624
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation, and 2 
       glycosites at Y208 and R274

<400>  38

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Lys 
        195                 200                 205             


Asp Gln Asn Ala Thr Lys Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp 
    210                 215                 220                 


Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys 
225                 230                 235                 240 


His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg Leu His Phe 
                245                 250                 255     


Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln Ala Cys His 
            260                 265                 270         


Leu Pro Leu Glu Ala Phe Thr Lys Asp Gln Asn Ala Thr Lys His Arg 
        275                 280                 285             


Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr Pro Val Gln 
    290                 295                 300                 


Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp Asn Gln Val 
305                 310                 315                 320 


Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser Gly Gly Asp 
                325                 330                 335     


Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg Leu Ala Leu 
            340                 345                 350         


Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln Gly Thr Gly 
        355                 360                 365             


Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val Ser Leu Thr Cys 
    370                 375                 380                 


Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser Gly Asp Ala 
385                 390                 395                 400 


Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu Gly Asp Gly 
                405                 410                 415     


Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp Thr Val Glu 
            420                 425                 430         


Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe 
        435                 440                 445             


Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val Phe 
    450                 455                 460                 


Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly 
465                 470                 475                 480 


Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln Asp 
                485                 490                 495     


Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu Leu Arg 
            500                 505                 510         


Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr Arg Thr Gly Leu 
        515                 520                 525             


Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu Arg Leu Ile Gly 
    530                 535                 540                 


His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly Pro Glu Glu Glu 
545                 550                 555                 560 


Gly Gly Arg Val Thr Ile Leu Gly Trp Pro Leu Ala Glu Arg Thr Val 
                565                 570                 575     


Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn Val Gly Gly Asp 
            580                 585                 590         


Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala Ile Ser Ala Leu 
        595                 600                 605             


Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg Glu Asp Leu Lys 
    610                 615                 620                 


<210>  39
<211>  630
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation, and 3 
       glycosites at Y208, R274 and A519

<400>  39

Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys Val 
1               5                   10                  15      


Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp Pro 
            20                  25                  30          


Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met Val 
        35                  40                  45              


Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala Leu 
    50                  55                  60                  


Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val Glu 
65                  70                  75                  80  


Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly Ser 
                85                  90                  95      


Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser Asn 
            100                 105                 110         


Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser His 
        115                 120                 125             


Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala Lys 
    130                 135                 140                 


Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn Glu 
145                 150                 155                 160 


Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val Met 
                165                 170                 175     


Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala Ser 
            180                 185                 190         


Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn Lys 
        195                 200                 205             


Asp Gln Asn Ala Thr Lys Leu Ala Gln Gln Arg Cys Asn Leu Asp Asp 
    210                 215                 220                 


Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn Pro Ala Lys 
225                 230                 235                 240 


His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg Leu His Phe 
                245                 250                 255     


Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln Ala Cys His 
            260                 265                 270         


Leu Pro Leu Glu Ala Phe Thr Lys Asp Gln Asn Ala Thr Lys His Arg 
        275                 280                 285             


Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr Pro Val Gln 
    290                 295                 300                 


Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp Asn Gln Val 
305                 310                 315                 320 


Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser Gly Gly Asp 
                325                 330                 335     


Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg Leu Ala Leu 
            340                 345                 350         


Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln Gly Thr Gly 
        355                 360                 365             


Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val Ser Leu Thr Cys 
    370                 375                 380                 


Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser Gly Asp Ala 
385                 390                 395                 400 


Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu Gly Asp Gly 
                405                 410                 415     


Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp Thr Val Glu 
            420                 425                 430         


Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe 
        435                 440                 445             


Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val Phe 
    450                 455                 460                 


Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly 
465                 470                 475                 480 


Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln Asp 
                485                 490                 495     


Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu Leu Arg 
            500                 505                 510         


Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr Arg Thr Gly Leu 
        515                 520                 525             


Thr Leu Lys Asp Gln Asn Ala Thr Lys Ala Pro Glu Ala Ala Gly Glu 
    530                 535                 540                 


Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile 
545                 550                 555                 560 


Thr Gly Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu Gly Trp Pro 
                565                 570                 575     


Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro 
            580                 585                 590         


Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu 
        595                 600                 605             


Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro 
    610                 615                 620                 


Pro Arg Glu Asp Leu Lys 
625                 630 


<210>  40
<211>  647
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation, and 4 
       glycosites at N-terminus, Y208, R274 and A519

<400>  40

Gly Ser Gly Gly Gly Asp Gln Asn Ala Thr Gly Ser Gly Gly Gly Lys 
1               5                   10                  15      


Leu Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys Ala Cys 
            20                  25                  30          


Val Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser Val Asp 
        35                  40                  45              


Pro Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr Ser Met 
    50                  55                  60                  


Val Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp Asn Ala 
65                  70                  75                  80  


Leu Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly Gly Val 
                85                  90                  95      


Glu Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala Arg Gly 
            100                 105                 110         


Ser Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys Pro Ser 
        115                 120                 125             


Asn Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln Leu Ser 
    130                 135                 140                 


His Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu Leu Ala 
145                 150                 155                 160 


Lys Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu Ser Asn 
                165                 170                 175     


Glu Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser Val Val 
            180                 185                 190         


Met Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu Trp Ala 
        195                 200                 205             


Ser Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val Tyr Asn 
    210                 215                 220                 


Lys Asp Gln Asn Ala Thr Lys Leu Ala Gln Gln Arg Cys Asn Leu Asp 
225                 230                 235                 240 


Asp Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn Pro Ala 
                245                 250                 255     


Lys His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg Leu His 
            260                 265                 270         


Phe Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln Ala Cys 
        275                 280                 285             


His Leu Pro Leu Glu Ala Phe Thr Lys Asp Gln Asn Ala Thr Lys His 
    290                 295                 300                 


Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr Pro Val 
305                 310                 315                 320 


Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp Asn Gln 
                325                 330                 335     


Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser Gly Gly 
            340                 345                 350         


Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg Leu Ala 
        355                 360                 365             


Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln Gly Thr 
    370                 375                 380                 


Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val Ser Leu Thr 
385                 390                 395                 400 


Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser Gly Asp 
                405                 410                 415     


Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu Gly Asp 
            420                 425                 430         


Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp Thr Val 
        435                 440                 445             


Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly Tyr Val 
    450                 455                 460                 


Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val 
465                 470                 475                 480 


Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg 
                485                 490                 495     


Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln 
            500                 505                 510         


Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu Leu 
        515                 520                 525             


Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr Arg Thr Gly 
    530                 535                 540                 


Leu Thr Leu Lys Asp Gln Asn Ala Thr Lys Ala Pro Glu Ala Ala Gly 
545                 550                 555                 560 


Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala 
                565                 570                 575     


Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu Gly Trp 
            580                 585                 590         


Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp 
        595                 600                 605             


Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys 
    610                 615                 620                 


Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys 
625                 630                 635                 640 


Pro Pro Arg Glu Asp Leu Lys 
                645         


<210>  41
<211>  653
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation, DsbA signal
       sequence, 3 glycosites at Y208, R274 and A519, and (HR)2 tag

<400>  41

Met Lys Lys Ile Trp Leu Ala Leu Ala Gly Leu Val Leu Ala Phe Ser 
1               5                   10                  15      


Ala Ser Ala Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys 
            20                  25                  30          


Ala Cys Val Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser 
        35                  40                  45              


Val Asp Pro Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr 
    50                  55                  60                  


Ser Met Val Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp 
65                  70                  75                  80  


Asn Ala Leu Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly 
                85                  90                  95      


Gly Val Glu Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala 
            100                 105                 110         


Arg Gly Ser Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys 
        115                 120                 125             


Pro Ser Asn Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln 
    130                 135                 140                 


Leu Ser His Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu 
145                 150                 155                 160 


Leu Ala Lys Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu 
                165                 170                 175     


Ser Asn Glu Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser 
            180                 185                 190         


Val Val Met Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu 
        195                 200                 205             


Trp Ala Ser Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val 
    210                 215                 220                 


Tyr Asn Lys Asp Gln Asn Ala Thr Lys Leu Ala Gln Gln Arg Cys Asn 
225                 230                 235                 240 


Leu Asp Asp Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn 
                245                 250                 255     


Pro Ala Lys His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg 
            260                 265                 270         


Leu His Phe Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln 
        275                 280                 285             


Ala Cys His Leu Pro Leu Glu Ala Phe Thr Lys Asp Gln Asn Ala Thr 
    290                 295                 300                 


Lys His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr 
305                 310                 315                 320 


Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp 
                325                 330                 335     


Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser 
            340                 345                 350         


Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg 
        355                 360                 365             


Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln 
    370                 375                 380                 


Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val Ser 
385                 390                 395                 400 


Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser 
                405                 410                 415     


Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu 
            420                 425                 430         


Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp 
        435                 440                 445             


Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly 
    450                 455                 460                 


Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser 
465                 470                 475                 480 


Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile 
                485                 490                 495     


Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr 
            500                 505                 510         


Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala 
        515                 520                 525             


Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr Arg 
    530                 535                 540                 


Thr Gly Leu Thr Leu Lys Asp Gln Asn Ala Thr Lys Ala Pro Glu Ala 
545                 550                 555                 560 


Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu 
                565                 570                 575     


Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu 
            580                 585                 590         


Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro 
        595                 600                 605             


Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro 
    610                 615                 620                 


Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro 
625                 630                 635                 640 


Gly Lys Pro Pro Arg Glu Asp Leu Lys His Arg His Arg 
                645                 650             


<210>  42
<211>  655
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation, DsbA signal
       sequence, 3 glycosites at Y208, R274 and A519, and (HR)3 tag

<400>  42

Met Lys Lys Ile Trp Leu Ala Leu Ala Gly Leu Val Leu Ala Phe Ser 
1               5                   10                  15      


Ala Ser Ala Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys 
            20                  25                  30          


Ala Cys Val Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser 
        35                  40                  45              


Val Asp Pro Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr 
    50                  55                  60                  


Ser Met Val Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp 
65                  70                  75                  80  


Asn Ala Leu Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly 
                85                  90                  95      


Gly Val Glu Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala 
            100                 105                 110         


Arg Gly Ser Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys 
        115                 120                 125             


Pro Ser Asn Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln 
    130                 135                 140                 


Leu Ser His Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu 
145                 150                 155                 160 


Leu Ala Lys Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu 
                165                 170                 175     


Ser Asn Glu Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser 
            180                 185                 190         


Val Val Met Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu 
        195                 200                 205             


Trp Ala Ser Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val 
    210                 215                 220                 


Tyr Asn Lys Asp Gln Asn Ala Thr Lys Leu Ala Gln Gln Arg Cys Asn 
225                 230                 235                 240 


Leu Asp Asp Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn 
                245                 250                 255     


Pro Ala Lys His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg 
            260                 265                 270         


Leu His Phe Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln 
        275                 280                 285             


Ala Cys His Leu Pro Leu Glu Ala Phe Thr Lys Asp Gln Asn Ala Thr 
    290                 295                 300                 


Lys His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr 
305                 310                 315                 320 


Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp 
                325                 330                 335     


Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser 
            340                 345                 350         


Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg 
        355                 360                 365             


Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln 
    370                 375                 380                 


Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val Ser 
385                 390                 395                 400 


Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser 
                405                 410                 415     


Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu 
            420                 425                 430         


Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp 
        435                 440                 445             


Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly 
    450                 455                 460                 


Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser 
465                 470                 475                 480 


Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile 
                485                 490                 495     


Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr 
            500                 505                 510         


Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala 
        515                 520                 525             


Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr Arg 
    530                 535                 540                 


Thr Gly Leu Thr Leu Lys Asp Gln Asn Ala Thr Lys Ala Pro Glu Ala 
545                 550                 555                 560 


Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu 
                565                 570                 575     


Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu 
            580                 585                 590         


Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro 
        595                 600                 605             


Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro 
    610                 615                 620                 


Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro 
625                 630                 635                 640 


Gly Lys Pro Pro Arg Glu Asp Leu Lys His Arg His Arg His Arg 
                645                 650                 655 


<210>  43
<211>  653
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation, DsbA signal
       sequence, 3 glycosites at Y208, R274 and A519, and R4 tag

<400>  43

Met Lys Lys Ile Trp Leu Ala Leu Ala Gly Leu Val Leu Ala Phe Ser 
1               5                   10                  15      


Ala Ser Ala Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys 
            20                  25                  30          


Ala Cys Val Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser 
        35                  40                  45              


Val Asp Pro Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr 
    50                  55                  60                  


Ser Met Val Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp 
65                  70                  75                  80  


Asn Ala Leu Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly 
                85                  90                  95      


Gly Val Glu Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala 
            100                 105                 110         


Arg Gly Ser Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys 
        115                 120                 125             


Pro Ser Asn Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln 
    130                 135                 140                 


Leu Ser His Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu 
145                 150                 155                 160 


Leu Ala Lys Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu 
                165                 170                 175     


Ser Asn Glu Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser 
            180                 185                 190         


Val Val Met Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu 
        195                 200                 205             


Trp Ala Ser Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val 
    210                 215                 220                 


Tyr Asn Lys Asp Gln Asn Ala Thr Lys Leu Ala Gln Gln Arg Cys Asn 
225                 230                 235                 240 


Leu Asp Asp Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn 
                245                 250                 255     


Pro Ala Lys His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg 
            260                 265                 270         


Leu His Phe Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln 
        275                 280                 285             


Ala Cys His Leu Pro Leu Glu Ala Phe Thr Lys Asp Gln Asn Ala Thr 
    290                 295                 300                 


Lys His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr 
305                 310                 315                 320 


Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp 
                325                 330                 335     


Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser 
            340                 345                 350         


Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg 
        355                 360                 365             


Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln 
    370                 375                 380                 


Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val Ser 
385                 390                 395                 400 


Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser 
                405                 410                 415     


Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu 
            420                 425                 430         


Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp 
        435                 440                 445             


Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly 
    450                 455                 460                 


Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser 
465                 470                 475                 480 


Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile 
                485                 490                 495     


Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr 
            500                 505                 510         


Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala 
        515                 520                 525             


Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr Arg 
    530                 535                 540                 


Thr Gly Leu Thr Leu Lys Asp Gln Asn Ala Thr Lys Ala Pro Glu Ala 
545                 550                 555                 560 


Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu 
                565                 570                 575     


Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu 
            580                 585                 590         


Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro 
        595                 600                 605             


Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro 
    610                 615                 620                 


Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro 
625                 630                 635                 640 


Gly Lys Pro Pro Arg Glu Asp Leu Lys Arg Arg Arg Arg 
                645                 650             


<210>  44
<211>  662
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation, DsbA signal
       sequence, 3 glycosites at Y208, R274 and A519, linker and (HR)4 
       tag

<400>  44

Met Lys Lys Ile Trp Leu Ala Leu Ala Gly Leu Val Leu Ala Phe Ser 
1               5                   10                  15      


Ala Ser Ala Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys 
            20                  25                  30          


Ala Cys Val Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser 
        35                  40                  45              


Val Asp Pro Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr 
    50                  55                  60                  


Ser Met Val Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp 
65                  70                  75                  80  


Asn Ala Leu Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly 
                85                  90                  95      


Gly Val Glu Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala 
            100                 105                 110         


Arg Gly Ser Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys 
        115                 120                 125             


Pro Ser Asn Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln 
    130                 135                 140                 


Leu Ser His Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu 
145                 150                 155                 160 


Leu Ala Lys Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu 
                165                 170                 175     


Ser Asn Glu Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser 
            180                 185                 190         


Val Val Met Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu 
        195                 200                 205             


Trp Ala Ser Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val 
    210                 215                 220                 


Tyr Asn Lys Asp Gln Asn Ala Thr Lys Leu Ala Gln Gln Arg Cys Asn 
225                 230                 235                 240 


Leu Asp Asp Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn 
                245                 250                 255     


Pro Ala Lys His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg 
            260                 265                 270         


Leu His Phe Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln 
        275                 280                 285             


Ala Cys His Leu Pro Leu Glu Ala Phe Thr Lys Asp Gln Asn Ala Thr 
    290                 295                 300                 


Lys His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr 
305                 310                 315                 320 


Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp 
                325                 330                 335     


Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser 
            340                 345                 350         


Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg 
        355                 360                 365             


Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln 
    370                 375                 380                 


Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val Ser 
385                 390                 395                 400 


Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser 
                405                 410                 415     


Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu 
            420                 425                 430         


Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp 
        435                 440                 445             


Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly 
    450                 455                 460                 


Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser 
465                 470                 475                 480 


Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile 
                485                 490                 495     


Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr 
            500                 505                 510         


Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala 
        515                 520                 525             


Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr Arg 
    530                 535                 540                 


Thr Gly Leu Thr Leu Lys Asp Gln Asn Ala Thr Lys Ala Pro Glu Ala 
545                 550                 555                 560 


Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu 
                565                 570                 575     


Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu 
            580                 585                 590         


Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro 
        595                 600                 605             


Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro 
    610                 615                 620                 


Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro 
625                 630                 635                 640 


Gly Lys Pro Pro Arg Glu Asp Leu Lys Gly Gly Ser Gly Gly His Arg 
                645                 650                 655     


His Arg His Arg His Arg 
            660         


<210>  45
<211>  660
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation, DsbA signal
       sequence, 3 glycosites at Y208, R274 and A519, linker and R6 tag

<400>  45

Met Lys Lys Ile Trp Leu Ala Leu Ala Gly Leu Val Leu Ala Phe Ser 
1               5                   10                  15      


Ala Ser Ala Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys 
            20                  25                  30          


Ala Cys Val Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser 
        35                  40                  45              


Val Asp Pro Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr 
    50                  55                  60                  


Ser Met Val Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp 
65                  70                  75                  80  


Asn Ala Leu Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly 
                85                  90                  95      


Gly Val Glu Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala 
            100                 105                 110         


Arg Gly Ser Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys 
        115                 120                 125             


Pro Ser Asn Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln 
    130                 135                 140                 


Leu Ser His Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu 
145                 150                 155                 160 


Leu Ala Lys Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu 
                165                 170                 175     


Ser Asn Glu Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser 
            180                 185                 190         


Val Val Met Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu 
        195                 200                 205             


Trp Ala Ser Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val 
    210                 215                 220                 


Tyr Asn Lys Asp Gln Asn Ala Thr Lys Leu Ala Gln Gln Arg Cys Asn 
225                 230                 235                 240 


Leu Asp Asp Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn 
                245                 250                 255     


Pro Ala Lys His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg 
            260                 265                 270         


Leu His Phe Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln 
        275                 280                 285             


Ala Cys His Leu Pro Leu Glu Ala Phe Thr Lys Asp Gln Asn Ala Thr 
    290                 295                 300                 


Lys His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr 
305                 310                 315                 320 


Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp 
                325                 330                 335     


Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser 
            340                 345                 350         


Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg 
        355                 360                 365             


Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln 
    370                 375                 380                 


Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val Ser 
385                 390                 395                 400 


Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser 
                405                 410                 415     


Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu 
            420                 425                 430         


Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp 
        435                 440                 445             


Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly 
    450                 455                 460                 


Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser 
465                 470                 475                 480 


Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile 
                485                 490                 495     


Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr 
            500                 505                 510         


Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala 
        515                 520                 525             


Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr Arg 
    530                 535                 540                 


Thr Gly Leu Thr Leu Lys Asp Gln Asn Ala Thr Lys Ala Pro Glu Ala 
545                 550                 555                 560 


Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu 
                565                 570                 575     


Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu 
            580                 585                 590         


Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro 
        595                 600                 605             


Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro 
    610                 615                 620                 


Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro 
625                 630                 635                 640 


Gly Lys Pro Pro Arg Glu Asp Leu Lys Gly Gly Ser Gly Gly Arg Arg 
                645                 650                 655     


Arg Arg Arg Arg 
            660 


<210>  46
<211>  661
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation, DsbA signal
       sequence, 3 glycosites at Y208, R274 and A519, and (PR)6 tag

<400>  46

Met Lys Lys Ile Trp Leu Ala Leu Ala Gly Leu Val Leu Ala Phe Ser 
1               5                   10                  15      


Ala Ser Ala Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys 
            20                  25                  30          


Ala Cys Val Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser 
        35                  40                  45              


Val Asp Pro Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr 
    50                  55                  60                  


Ser Met Val Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp 
65                  70                  75                  80  


Asn Ala Leu Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly 
                85                  90                  95      


Gly Val Glu Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala 
            100                 105                 110         


Arg Gly Ser Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys 
        115                 120                 125             


Pro Ser Asn Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln 
    130                 135                 140                 


Leu Ser His Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu 
145                 150                 155                 160 


Leu Ala Lys Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu 
                165                 170                 175     


Ser Asn Glu Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser 
            180                 185                 190         


Val Val Met Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu 
        195                 200                 205             


Trp Ala Ser Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val 
    210                 215                 220                 


Tyr Asn Lys Asp Gln Asn Ala Thr Lys Leu Ala Gln Gln Arg Cys Asn 
225                 230                 235                 240 


Leu Asp Asp Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn 
                245                 250                 255     


Pro Ala Lys His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg 
            260                 265                 270         


Leu His Phe Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln 
        275                 280                 285             


Ala Cys His Leu Pro Leu Glu Ala Phe Thr Lys Asp Gln Asn Ala Thr 
    290                 295                 300                 


Lys His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr 
305                 310                 315                 320 


Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp 
                325                 330                 335     


Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser 
            340                 345                 350         


Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg 
        355                 360                 365             


Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln 
    370                 375                 380                 


Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val Ser 
385                 390                 395                 400 


Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser 
                405                 410                 415     


Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu 
            420                 425                 430         


Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp 
        435                 440                 445             


Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly 
    450                 455                 460                 


Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser 
465                 470                 475                 480 


Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile 
                485                 490                 495     


Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr 
            500                 505                 510         


Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala 
        515                 520                 525             


Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr Arg 
    530                 535                 540                 


Thr Gly Leu Thr Leu Lys Asp Gln Asn Ala Thr Lys Ala Pro Glu Ala 
545                 550                 555                 560 


Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu 
                565                 570                 575     


Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu 
            580                 585                 590         


Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro 
        595                 600                 605             


Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro 
    610                 615                 620                 


Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro 
625                 630                 635                 640 


Gly Lys Pro Pro Arg Glu Asp Leu Lys Pro Arg Pro Arg Pro Arg Pro 
                645                 650                 655     


Arg Pro Arg Pro Arg 
            660     


<210>  47
<211>  661
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of EPA with detoxifying mutation, DsbA signal
       sequence, 3 glycosites at Y208, R274 and A519, and (PSR)4 tag

<400>  47

Met Lys Lys Ile Trp Leu Ala Leu Ala Gly Leu Val Leu Ala Phe Ser 
1               5                   10                  15      


Ala Ser Ala Ala Glu Glu Ala Phe Asp Leu Trp Asn Glu Cys Ala Lys 
            20                  25                  30          


Ala Cys Val Leu Asp Leu Lys Asp Gly Val Arg Ser Ser Arg Met Ser 
        35                  40                  45              


Val Asp Pro Ala Ile Ala Asp Thr Asn Gly Gln Gly Val Leu His Tyr 
    50                  55                  60                  


Ser Met Val Leu Glu Gly Gly Asn Asp Ala Leu Lys Leu Ala Ile Asp 
65                  70                  75                  80  


Asn Ala Leu Ser Ile Thr Ser Asp Gly Leu Thr Ile Arg Leu Glu Gly 
                85                  90                  95      


Gly Val Glu Pro Asn Lys Pro Val Arg Tyr Ser Tyr Thr Arg Gln Ala 
            100                 105                 110         


Arg Gly Ser Trp Ser Leu Asn Trp Leu Val Pro Ile Gly His Glu Lys 
        115                 120                 125             


Pro Ser Asn Ile Lys Val Phe Ile His Glu Leu Asn Ala Gly Asn Gln 
    130                 135                 140                 


Leu Ser His Met Ser Pro Ile Tyr Thr Ile Glu Met Gly Asp Glu Leu 
145                 150                 155                 160 


Leu Ala Lys Leu Ala Arg Asp Ala Thr Phe Phe Val Arg Ala His Glu 
                165                 170                 175     


Ser Asn Glu Met Gln Pro Thr Leu Ala Ile Ser His Ala Gly Val Ser 
            180                 185                 190         


Val Val Met Ala Gln Ala Gln Pro Arg Arg Glu Lys Arg Trp Ser Glu 
        195                 200                 205             


Trp Ala Ser Gly Lys Val Leu Cys Leu Leu Asp Pro Leu Asp Gly Val 
    210                 215                 220                 


Tyr Asn Lys Asp Gln Asn Ala Thr Lys Leu Ala Gln Gln Arg Cys Asn 
225                 230                 235                 240 


Leu Asp Asp Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn 
                245                 250                 255     


Pro Ala Lys His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg 
            260                 265                 270         


Leu His Phe Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln 
        275                 280                 285             


Ala Cys His Leu Pro Leu Glu Ala Phe Thr Lys Asp Gln Asn Ala Thr 
    290                 295                 300                 


Lys His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr 
305                 310                 315                 320 


Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp 
                325                 330                 335     


Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser 
            340                 345                 350         


Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg 
        355                 360                 365             


Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln 
    370                 375                 380                 


Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Ala Asp Val Val Ser 
385                 390                 395                 400 


Leu Thr Cys Pro Val Ala Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser 
                405                 410                 415     


Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu 
            420                 425                 430         


Gly Asp Gly Gly Asp Val Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp 
        435                 440                 445             


Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly 
    450                 455                 460                 


Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser 
465                 470                 475                 480 


Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile 
                485                 490                 495     


Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr 
            500                 505                 510         


Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala 
        515                 520                 525             


Leu Leu Arg Val Tyr Val Pro Arg Trp Ser Leu Pro Gly Phe Tyr Arg 
    530                 535                 540                 


Thr Gly Leu Thr Leu Lys Asp Gln Asn Ala Thr Lys Ala Pro Glu Ala 
545                 550                 555                 560 


Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu 
                565                 570                 575     


Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Val Thr Ile Leu 
            580                 585                 590         


Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro 
        595                 600                 605             


Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro 
    610                 615                 620                 


Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro 
625                 630                 635                 640 


Gly Lys Pro Pro Arg Glu Asp Leu Lys Pro Ser Arg Pro Ser Arg Pro 
                645                 650                 655     


Ser Arg Pro Ser Arg 
            660     


