                         SEQUENCE LISTING

<110>  ew nutrition GmbH
 
<120>  Stable synthetic phytase variants

<130>  ED41239

<160>  5     

<170>  PatentIn version 3.5

<210>  1
<211>  413
<212>  PRT
<213>  Hafnia alvei

<400>  1

Ser Asp Thr Ala Pro Ala Gly Phe Gln Leu Glu Lys Val Val Ile Leu 
1               5                   10                  15      


Ser Arg His Gly Val Arg Ala Pro Thr Lys Met Thr Gln Thr Met Arg 
            20                  25                  30          


Asn Val Thr Pro His Gln Trp Pro Glu Trp Pro Val Lys Leu Gly Tyr 
        35                  40                  45              


Ile Thr Pro Arg Gly Glu His Leu Ile Ser Leu Met Gly Gly Phe Tyr 
    50                  55                  60                  


Arg Glu Arg Phe Gln Gln Gln Gly Leu Leu Pro Lys Asp Asn Cys Pro 
65                  70                  75                  80  


Thr Pro Asp Ala Val Tyr Val Trp Ala Asp Val Asp Gln Arg Thr Arg 
                85                  90                  95      


Lys Thr Gly Glu Ala Phe Leu Ala Gly Leu Ala Pro Gln Cys Asp Leu 
            100                 105                 110         


Ala Ile His His Gln Gln Asn Ile Gln Gln Ala Asp Pro Leu Phe His 
        115                 120                 125             


Pro Val Lys Ala Gly Ile Cys Ser Met Asp Lys Ser Gln Ala His Ala 
    130                 135                 140                 


Ala Val Glu Lys Gln Ala Gly Thr Pro Ile Glu Thr Leu Asn Gln Arg 
145                 150                 155                 160 


Tyr Gln Ala Ser Leu Ala Leu Met Ser Ser Val Leu Asp Phe Pro Lys 
                165                 170                 175     


Ser Pro Tyr Cys Gln Gln His Asn Ile Gly Lys Leu Cys Asp Phe Ser 
            180                 185                 190         


Gln Ala Met Pro Ser Arg Leu Ala Ile Asn Asp Asp Gly Asn Lys Val 
        195                 200                 205             


Ala Leu Glu Gly Ala Val Gly Leu Ser Ser Thr Leu Ala Glu Ile Phe 
    210                 215                 220                 


Leu Leu Glu His Ala Gln Gly Met Pro Lys Val Ala Trp Gly Asn Ile 
225                 230                 235                 240 


His Thr Glu Gln Gln Trp Asp Ser Leu Leu Lys Leu His Asn Ala Gln 
                245                 250                 255     


Phe Asp Leu Met Ser Arg Thr Pro Tyr Ile Ala Lys His Asn Gly Thr 
            260                 265                 270         


Pro Leu Leu Gln Thr Ile Ala His Ala Leu Gly Ser Asn Ile Ala Ser 
        275                 280                 285             


Arg Pro Leu Pro Asp Ile Ser Pro Asp Asn Lys Ile Leu Phe Ile Ala 
    290                 295                 300                 


Gly His Asp Thr Asn Ile Ala Asn Ile Ser Gly Met Leu Gly Met Thr 
305                 310                 315                 320 


Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Ala Leu 
                325                 330                 335     


Val Phe Glu Arg Trp Val Asp Asn Ala Gly Lys Pro Tyr Val Ser Val 
            340                 345                 350         


Asn Met Val Tyr Gln Thr Leu Ala Gln Leu His Asp Gln Thr Pro Leu 
        355                 360                 365             


Thr Leu Gln His Pro Ala Gly Ser Val Arg Leu Asn Ile Pro Gly Cys 
    370                 375                 380                 


Ser Asp Gln Thr Pro Asp Gly Tyr Cys Pro Leu Ser Thr Phe Ser Arg 
385                 390                 395                 400 


Leu Val Asn His Ser Val Glu Pro Ala Cys Gln Leu Pro 
                405                 410             


<210>  2
<211>  411
<212>  PRT
<213>  Obesumbacterium

<400>  2

Ser Glu Thr Val Pro Ser Gly Tyr Gln Leu Glu Lys Val Val Ile Leu 
1               5                   10                  15      


Ser Arg His Gly Val Arg Ala Pro Thr Lys Met Thr Gln Thr Met Arg 
            20                  25                  30          


Asp Val Thr Pro Asn Thr Trp Pro Glu Trp Pro Val Lys Leu Gly Tyr 
        35                  40                  45              


Ile Thr Pro Arg Gly Glu His Leu Val Ser Leu Met Gly Gly Phe Tyr 
    50                  55                  60                  


Arg Gln Lys Phe Gln Gln Leu Gly Ile Leu Ser Lys Asp Arg Cys Pro 
65                  70                  75                  80  


Thr Ala Asn Asp Val Tyr Val Trp Ala Asp Val Asp Gln Arg Thr Arg 
                85                  90                  95      


Lys Thr Gly Glu Ala Phe Leu Ala Gly Leu Ala Pro Glu Cys His Leu 
            100                 105                 110         


Ser Ile His His Gln Gln Asp Ile Lys Gln Val Asp Ser Leu Phe His 
        115                 120                 125             


Pro Val Lys Ala Gly Val Cys Thr Met Glu Lys Ala Gln Val Gln Gln 
    130                 135                 140                 


Ala Val Glu Gln Gln Ala Gly Met Pro Ile Ala Gln Leu Asn Gln His 
145                 150                 155                 160 


Tyr Arg Pro Ala Leu Ala Leu Met Ser Asn Val Leu Asn Phe Pro Thr 
                165                 170                 175     


Ser Ala Tyr Cys Gln Gln His Ser Ala Glu Gln Thr Cys Asn Phe Ala 
            180                 185                 190         


Glu Val Met Pro Ser Lys Leu Ser Ile Lys Asp Asp Gly Asn Lys Val 
        195                 200                 205             


Ala Leu Asp Gly Ala Val Gly Leu Ser Ser Thr Leu Ala Glu Ile Phe 
    210                 215                 220                 


Leu Leu Glu His Ala Gln Gly Met Pro Asn Ala Ala Trp Gly Lys Ile 
225                 230                 235                 240 


His Ser Glu Gln Asp Trp Asn Thr Leu Leu Thr Leu His Asn Ala Gln 
                245                 250                 255     


Phe Asp Leu Met Ser Arg Thr Pro Tyr Ile Ala Lys His Asn Gly Thr 
            260                 265                 270         


Pro Leu Leu Gln Thr Ile Val Ser Ala Ile Asn Ser Gln Thr Ser Pro 
        275                 280                 285             


Arg Glu Leu Pro Glu Leu Ser Ala Asp Asn Lys Ile Leu Phe Leu Ala 
    290                 295                 300                 


Gly His Asp Thr Asn Ile Ala Asn Ile Ala Gly Met Leu Gly Leu Ser 
305                 310                 315                 320 


Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Ala Leu 
                325                 330                 335     


Val Phe Glu Arg Trp Ser Asp Lys Thr Gly Lys Lys Tyr Val Ser Ala 
            340                 345                 350         


Gln Met Met Tyr Gln Thr Leu Ala Gln Leu Arg Asn Gln Thr Pro Leu 
        355                 360                 365             


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


Asp Asp Gln Thr Ala Glu Gly Tyr Cys Pro Leu Asp Thr Phe Thr Arg 
385                 390                 395                 400 


Leu Ala Lys Gln Asn Glu Leu Ala Glu Cys Gln 
                405                 410     


<210>  3
<211>  411
<212>  PRT
<213>  artificial sequence

<220>
<223>  shuffled variant

<400>  3

Ser Glu Thr Val Pro Ser Gly Tyr Gln Leu Glu Lys Val Val Ile Leu 
1               5                   10                  15      


Ser Arg His Gly Val Arg Ala Pro Thr Lys Met Thr Gln Thr Met Arg 
            20                  25                  30          


Asp Val Thr Pro Asn Thr Trp Pro Glu Trp Pro Val Lys Leu Gly Tyr 
        35                  40                  45              


Ile Thr Pro Arg Gly Glu His Leu Val Ser Leu Met Gly Gly Phe Tyr 
    50                  55                  60                  


Arg Gln Lys Phe Gln Gln Leu Gly Ile Leu Ser Lys Asp Arg Cys Pro 
65                  70                  75                  80  


Thr Ala Asn Asp Val Tyr Val Trp Ala Asp Val Asp Gln Arg Thr Arg 
                85                  90                  95      


Lys Thr Gly Glu Ala Phe Leu Ala Gly Leu Ala Pro Gln Cys Asp Leu 
            100                 105                 110         


Ala Ile His His Gln Gln Asn Ile Gln Gln Ala Asp Pro Leu Phe His 
        115                 120                 125             


Pro Val Lys Ala Gly Ile Cys Ser Met Asp Lys Ser Gln Ala His Ala 
    130                 135                 140                 


Ala Val Glu Lys Gln Ala Gly Thr Pro Ile Glu Thr Leu Asn Gln Arg 
145                 150                 155                 160 


Tyr Gln Ala Ser Leu Ala Leu Met Ser Ser Val Leu Asp Phe Pro Lys 
                165                 170                 175     


Ser Pro Tyr Cys Gln Gln His Asn Ile Gly Lys Leu Cys Asp Phe Ser 
            180                 185                 190         


Gln Ala Met Pro Ser Arg Leu Ala Ile Asn Asp Asp Gly Asn Lys Val 
        195                 200                 205             


Ala Leu Glu Gly Ala Val Gly Leu Ser Ser Thr Leu Ala Glu Ile Phe 
    210                 215                 220                 


Leu Leu Glu His Ala Gln Gly Met Pro Lys Val Ala Trp Gly Asn Ile 
225                 230                 235                 240 


His Thr Glu Gln Gln Trp Asp Ser Leu Leu Lys Leu His Asn Ala Gln 
                245                 250                 255     


Phe Asp Leu Met Ser Arg Thr Pro Tyr Ile Ala Lys His Asn Gly Thr 
            260                 265                 270         


Pro Leu Leu Gln Thr Ile Val Ser Ala Ile Asn Ser Gln Thr Ser Pro 
        275                 280                 285             


Arg Glu Leu Pro Glu Leu Ser Ala Asp Asn Lys Ile Leu Phe Leu Ala 
    290                 295                 300                 


Gly His Asp Thr Asn Ile Ala Asn Ile Ala Gly Met Leu Gly Leu Ser 
305                 310                 315                 320 


Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Ala Leu 
                325                 330                 335     


Val Phe Glu Arg Trp Ser Asp Lys Thr Gly Lys Lys Tyr Val Ser Ala 
            340                 345                 350         


Gln Met Met Tyr Gln Thr Leu Ala Gln Leu Arg Asn Gln Thr Pro Leu 
        355                 360                 365             


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


Asp Asp Gln Thr Ala Glu Gly Tyr Cys Pro Leu Asp Thr Phe Thr Arg 
385                 390                 395                 400 


Leu Ala Lys Gln Asn Glu Leu Ala Glu Cys Gln 
                405                 410     


<210>  4
<211>  411
<212>  PRT
<213>  artificial sequence

<220>
<223>  shuffled variant

<400>  4

Ser Asp Thr Ala Pro Ala Gly Phe Gln Leu Glu Lys Val Val Ile Leu 
1               5                   10                  15      


Ser Arg His Gly Val Arg Ala Pro Thr Lys Met Thr Gln Thr Met Arg 
            20                  25                  30          


Asn Val Thr Pro His Gln Trp Pro Glu Trp Pro Val Lys Leu Gly Tyr 
        35                  40                  45              


Ile Thr Pro Arg Gly Glu His Leu Ile Ser Leu Met Gly Gly Phe Tyr 
    50                  55                  60                  


Arg Glu Arg Phe Gln Gln Gln Gly Leu Leu Pro Lys Asp Asn Cys Pro 
65                  70                  75                  80  


Thr Pro Asp Ala Val Tyr Val Trp Ala Asp Val Asp Gln Arg Thr Arg 
                85                  90                  95      


Lys Thr Gly Glu Ala Phe Leu Ala Gly Leu Ala Pro Gln Cys Asp Leu 
            100                 105                 110         


Ala Ile His His Gln Gln Asn Ile Gln Gln Ala Asp Pro Leu Phe His 
        115                 120                 125             


Pro Val Lys Ala Gly Ile Cys Ser Met Asp Lys Ser Gln Ala His Ala 
    130                 135                 140                 


Ala Val Glu Lys Gln Ala Gly Thr Pro Ile Glu Thr Leu Asn Gln Arg 
145                 150                 155                 160 


Tyr Gln Ala Ser Leu Ala Leu Met Ser Ser Val Leu Asp Phe Pro Lys 
                165                 170                 175     


Ser Pro Tyr Cys Gln Gln His Asn Ile Gly Lys Leu Cys Asp Phe Ser 
            180                 185                 190         


Gln Ala Met Pro Ser Arg Leu Ala Ile Asn Asp Asp Gly Asn Lys Val 
        195                 200                 205             


Ala Leu Glu Gly Ala Val Gly Leu Ser Ser Thr Leu Ala Glu Ile Phe 
    210                 215                 220                 


Leu Leu Glu His Ala Gln Gly Met Pro Lys Val Ala Trp Gly Asn Ile 
225                 230                 235                 240 


His Thr Glu Gln Gln Trp Asp Ser Leu Leu Lys Leu His Asn Ala Gln 
                245                 250                 255     


Phe Asp Leu Met Ser Arg Thr Pro Tyr Ile Ala Lys His Asn Gly Thr 
            260                 265                 270         


Pro Leu Leu Gln Thr Ile Val Ser Ala Ile Asn Ser Gln Thr Ser Pro 
        275                 280                 285             


Arg Glu Leu Pro Glu Leu Ser Ala Asp Asn Lys Ile Leu Phe Leu Ala 
    290                 295                 300                 


Gly His Asp Thr Asn Ile Ala Asn Ile Ala Gly Met Leu Gly Leu Ser 
305                 310                 315                 320 


Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Ala Leu 
                325                 330                 335     


Val Phe Glu Arg Trp Ser Asp Lys Thr Gly Lys Lys Tyr Val Ser Ala 
            340                 345                 350         


Gln Met Met Tyr Gln Thr Leu Ala Gln Leu Arg Asn Gln Thr Pro Leu 
        355                 360                 365             


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


Asp Asp Gln Thr Ala Glu Gly Tyr Cys Pro Leu Asp Thr Phe Thr Arg 
385                 390                 395                 400 


Leu Ala Lys Gln Asn Glu Leu Ala Glu Cys Gln 
                405                 410     


<210>  5
<211>  413
<212>  PRT
<213>  artificial sequence

<220>
<223>  shuffled variant

<400>  5

Ser Glu Thr Val Pro Ser Gly Tyr Gln Leu Glu Lys Val Val Ile Leu 
1               5                   10                  15      


Ser Arg His Gly Val Arg Ala Pro Thr Lys Met Thr Gln Thr Met Arg 
            20                  25                  30          


Asp Val Thr Pro Asn Thr Trp Pro Glu Trp Pro Val Lys Leu Gly Tyr 
        35                  40                  45              


Ile Thr Pro Arg Gly Glu His Leu Val Ser Leu Met Gly Gly Phe Tyr 
    50                  55                  60                  


Arg Gln Lys Phe Gln Gln Leu Gly Ile Leu Ser Lys Asp Arg Cys Pro 
65                  70                  75                  80  


Thr Ala Asn Asp Val Tyr Val Trp Ala Asp Val Asp Gln Arg Thr Arg 
                85                  90                  95      


Lys Thr Gly Glu Ala Phe Leu Ala Gly Leu Ala Pro Gln Cys Asp Leu 
            100                 105                 110         


Ala Ile His His Gln Gln Asn Ile Gln Gln Ala Asp Pro Leu Phe His 
        115                 120                 125             


Pro Val Lys Ala Gly Ile Cys Ser Met Asp Lys Ser Gln Ala His Ala 
    130                 135                 140                 


Ala Val Glu Lys Gln Ala Gly Thr Pro Ile Glu Thr Leu Asn Gln Arg 
145                 150                 155                 160 


Tyr Gln Ala Ser Leu Ala Leu Met Ser Ser Val Leu Asp Phe Pro Lys 
                165                 170                 175     


Ser Pro Tyr Cys Gln Gln His Asn Ile Gly Lys Leu Cys Asp Phe Ser 
            180                 185                 190         


Gln Ala Met Pro Ser Arg Leu Ala Ile Asn Asp Asp Gly Asn Lys Val 
        195                 200                 205             


Ala Leu Glu Gly Ala Val Gly Leu Ser Ser Thr Leu Ala Glu Ile Phe 
    210                 215                 220                 


Leu Leu Glu His Ala Gln Gly Met Pro Lys Val Ala Trp Gly Asn Ile 
225                 230                 235                 240 


His Thr Glu Gln Gln Trp Asp Ser Leu Leu Lys Leu His Asn Ala Gln 
                245                 250                 255     


Phe Asp Leu Met Ser Arg Thr Pro Tyr Ile Ala Lys His Asn Gly Thr 
            260                 265                 270         


Pro Leu Leu Gln Thr Ile Ala His Ala Leu Gly Ser Asn Ile Ala Ser 
        275                 280                 285             


Arg Pro Leu Pro Asp Ile Ser Pro Asp Asn Lys Ile Leu Phe Ile Ala 
    290                 295                 300                 


Gly His Asp Thr Asn Ile Ala Asn Ile Ser Gly Met Leu Gly Met Thr 
305                 310                 315                 320 


Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Ala Leu 
                325                 330                 335     


Val Phe Glu Arg Trp Val Asp Asn Ala Gly Lys Pro Tyr Val Ser Val 
            340                 345                 350         


Asn Met Val Tyr Gln Thr Leu Ala Gln Leu His Asp Gln Thr Pro Leu 
        355                 360                 365             


Thr Leu Gln His Pro Ala Gly Ser Val Arg Leu Asn Ile Pro Gly Cys 
    370                 375                 380                 


Ser Asp Gln Thr Pro Asp Gly Tyr Cys Pro Leu Ser Thr Phe Ser Arg 
385                 390                 395                 400 


Leu Val Asn His Ser Val Glu Pro Ala Cys Gln Leu Pro 
                405                 410             


