                         SEQUENCE LISTING

<110>  ew nutrition
 
<120>  Stable protease variants

<130>  ED41048

<140>  16206367.1 - 1410
<141>  22.12.2016

<160>  5     

<170>  PatentIn version 3.5

<210>  1
<211>  552
<212>  PRT
<213>  artificial

<220>
<223>  Kumamolisin 1 proenzyme, backbone variant (N-terminal M is 
       lacking)

<400>  1

Ser Asp Met Glu Lys Pro Trp Lys Glu Gly Glu Glu Ala Arg Ala Val 
1               5                   10                  15      


Leu Gln Gly His Ala Arg Ala Gln Ala Pro Gln Ala Val Asp Lys Gly 
            20                  25                  30          


Pro Val Ala Gly Asp Glu Arg Met Ala Val Thr Val Val Leu Arg Arg 
        35                  40                  45              


Gln Arg Ala Gly Glu Leu Ala Ala His Val Glu Arg Gln Ala Ala Ile 
    50                  55                  60                  


Ala Pro His Ala Arg Glu His Leu Lys Arg Glu Ala Phe Ala Ala Ser 
65                  70                  75                  80  


His Gly Ala Ser Leu Asp Asp Phe Ala Glu Leu Arg Arg Phe Ala Asp 
                85                  90                  95      


Ala His Gly Leu Ala Leu Asp Arg Ala Asn Val Ala Ala Gly Thr Ala 
            100                 105                 110         


Val Leu Ser Gly Pro Val Asp Ala Ile Asn Arg Ala Phe Gly Val Glu 
        115                 120                 125             


Leu Arg His Phe Asp His Pro Asp Gly Ser Tyr Arg Ser Tyr Leu Gly 
    130                 135                 140                 


Glu Val Thr Val Pro Ala Ser Ile Ala Pro Met Ile Glu Ala Val Leu 
145                 150                 155                 160 


Gly Leu Asp Thr Arg Pro Val Ala Arg Pro His Phe Arg Met Gln Arg 
                165                 170                 175     


Arg Ala Glu Gly Gly Phe Glu Ala Arg Ser Gln Ala Ala Ala Pro Thr 
            180                 185                 190         


Ala Tyr Thr Pro Leu Asp Val Ala Gln Ala Tyr Gln Phe Pro Glu Gly 
        195                 200                 205             


Leu Asp Gly Gln Gly Gln Cys Ile Ala Ile Ile Glu Leu Gly Gly Gly 
    210                 215                 220                 


Tyr Asp Glu Ala Ser Leu Ala Gln Tyr Phe Ala Ser Leu Gly Val Pro 
225                 230                 235                 240 


Ala Pro Gln Val Val Ser Val Ser Val Asp Gly Ala Ser Asn Gln Pro 
                245                 250                 255     


Thr Gly Asp Pro Ser Gly Pro Asp Gly Glu Val Glu Leu Asp Ile Glu 
            260                 265                 270         


Val Ala Gly Ala Leu Ala Pro Gly Ala Lys Phe Ala Val Tyr Phe Ala 
        275                 280                 285             


Pro Asn Thr Asp Ala Gly Phe Leu Asp Ala Ile Thr Thr Ala Ile His 
    290                 295                 300                 


Asp Pro Thr Leu Lys Pro Ser Val Val Ser Ile Ser Trp Gly Gly Pro 
305                 310                 315                 320 


Glu Asp Ser Trp Thr Ser Ala Ala Ile Ala Ala Met Asn Arg Ala Phe 
                325                 330                 335     


Leu Asp Ala Ala Ala Leu Gly Val Thr Val Leu Ala Ala Ala Gly Asp 
            340                 345                 350         


Ser Gly Ser Thr Asp Gly Glu Gln Asp Gly Leu Tyr His Val Asp Phe 
        355                 360                 365             


Pro Ala Ala Ser Pro Tyr Val Leu Ala Cys Gly Gly Thr Arg Leu Val 
    370                 375                 380                 


Ala Ser Gly Gly Arg Ile Ala Gln Glu Thr Val Trp Asn Asp Gly Pro 
385                 390                 395                 400 


Asp Gly Gly Ala Thr Gly Gly Gly Val Ser Arg Ile Phe Pro Leu Pro 
                405                 410                 415     


Ala Trp Gln Glu His Ala Asn Val Pro Pro Ser Ala Asn Pro Gly Ala 
            420                 425                 430         


Ser Ser Gly Arg Gly Val Pro Asp Leu Ala Gly Asn Ala Asp Pro Ala 
        435                 440                 445             


Thr Gly Tyr Glu Val Val Ile Asp Gly Glu Ala Thr Val Ile Gly Gly 
    450                 455                 460                 


Thr Ser Ala Val Ala Pro Leu Phe Ala Ala Leu Val Ala Arg Ile Asn 
465                 470                 475                 480 


Gln Lys Leu Gly Lys Ala Val Gly Tyr Leu Asn Pro Thr Leu Tyr Gln 
                485                 490                 495     


Leu Pro Ala Asp Val Phe His Asp Ile Thr Glu Gly Asn Asn Asp Ile 
            500                 505                 510         


Ala Asn Arg Ala Gln Ile Tyr Gln Ala Gly Pro Gly Trp Asp Pro Cys 
        515                 520                 525             


Thr Gly Leu Gly Ser Pro Ile Gly Val Arg Leu Leu Gln Ala Leu Leu 
    530                 535                 540                 


Pro Ser Ala Ser Gln Pro Gln Pro 
545                 550         


<210>  2
<211>  587
<212>  PRT
<213>  artificial

<220>
<223>  Kumamolisin 1 proenzyme backbone variant with leader sequence and
       His tag

<400>  2

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


Thr Ala Leu Leu Ala Gly Gly Ala Thr Gln Ala Phe Ala Ser Asp Met 
            20                  25                  30          


Glu Lys Pro Trp Lys Glu Gly Glu Glu Ala Arg Ala Val Leu Gln Gly 
        35                  40                  45              


His Ala Arg Ala Gln Ala Pro Gln Ala Val Asp Lys Gly Pro Val Ala 
    50                  55                  60                  


Gly Asp Glu Arg Met Ala Val Thr Val Val Leu Arg Arg Gln Arg Ala 
65                  70                  75                  80  


Gly Glu Leu Ala Ala His Val Glu Arg Gln Ala Ala Ile Ala Pro His 
                85                  90                  95      


Ala Arg Glu His Leu Lys Arg Glu Ala Phe Ala Ala Ser His Gly Ala 
            100                 105                 110         


Ser Leu Asp Asp Phe Ala Glu Leu Arg Arg Phe Ala Asp Ala His Gly 
        115                 120                 125             


Leu Ala Leu Asp Arg Ala Asn Val Ala Ala Gly Thr Ala Val Leu Ser 
    130                 135                 140                 


Gly Pro Val Asp Ala Ile Asn Arg Ala Phe Gly Val Glu Leu Arg His 
145                 150                 155                 160 


Phe Asp His Pro Asp Gly Ser Tyr Arg Ser Tyr Leu Gly Glu Val Thr 
                165                 170                 175     


Val Pro Ala Ser Ile Ala Pro Met Ile Glu Ala Val Leu Gly Leu Asp 
            180                 185                 190         


Thr Arg Pro Val Ala Arg Pro His Phe Arg Met Gln Arg Arg Ala Glu 
        195                 200                 205             


Gly Gly Phe Glu Ala Arg Ser Gln Ala Ala Ala Pro Thr Ala Tyr Thr 
    210                 215                 220                 


Pro Leu Asp Val Ala Gln Ala Tyr Gln Phe Pro Glu Gly Leu Asp Gly 
225                 230                 235                 240 


Gln Gly Gln Cys Ile Ala Ile Ile Glu Leu Gly Gly Gly Tyr Asp Glu 
                245                 250                 255     


Ala Ser Leu Ala Gln Tyr Phe Ala Ser Leu Gly Val Pro Ala Pro Gln 
            260                 265                 270         


Val Val Ser Val Ser Val Asp Gly Ala Ser Asn Gln Pro Thr Gly Asp 
        275                 280                 285             


Pro Ser Gly Pro Asp Gly Glu Val Glu Leu Asp Ile Glu Val Ala Gly 
    290                 295                 300                 


Ala Leu Ala Pro Gly Ala Lys Phe Ala Val Tyr Phe Ala Pro Asn Thr 
305                 310                 315                 320 


Asp Ala Gly Phe Leu Asp Ala Ile Thr Thr Ala Ile His Asp Pro Thr 
                325                 330                 335     


Leu Lys Pro Ser Val Val Ser Ile Ser Trp Gly Gly Pro Glu Asp Ser 
            340                 345                 350         


Trp Thr Ser Ala Ala Ile Ala Ala Met Asn Arg Ala Phe Leu Asp Ala 
        355                 360                 365             


Ala Ala Leu Gly Val Thr Val Leu Ala Ala Ala Gly Asp Ser Gly Ser 
    370                 375                 380                 


Thr Asp Gly Glu Gln Asp Gly Leu Tyr His Val Asp Phe Pro Ala Ala 
385                 390                 395                 400 


Ser Pro Tyr Val Leu Ala Cys Gly Gly Thr Arg Leu Val Ala Ser Gly 
                405                 410                 415     


Gly Arg Ile Ala Gln Glu Thr Val Trp Asn Asp Gly Pro Asp Gly Gly 
            420                 425                 430         


Ala Thr Gly Gly Gly Val Ser Arg Ile Phe Pro Leu Pro Ala Trp Gln 
        435                 440                 445             


Glu His Ala Asn Val Pro Pro Ser Ala Asn Pro Gly Ala Ser Ser Gly 
    450                 455                 460                 


Arg Gly Val Pro Asp Leu Ala Gly Asn Ala Asp Pro Ala Thr Gly Tyr 
465                 470                 475                 480 


Glu Val Val Ile Asp Gly Glu Ala Thr Val Ile Gly Gly Thr Ser Ala 
                485                 490                 495     


Val Ala Pro Leu Phe Ala Ala Leu Val Ala Arg Ile Asn Gln Lys Leu 
            500                 505                 510         


Gly Lys Ala Val Gly Tyr Leu Asn Pro Thr Leu Tyr Gln Leu Pro Ala 
        515                 520                 525             


Asp Val Phe His Asp Ile Thr Glu Gly Asn Asn Asp Ile Ala Asn Arg 
    530                 535                 540                 


Ala Gln Ile Tyr Gln Ala Gly Pro Gly Trp Asp Pro Cys Thr Gly Leu 
545                 550                 555                 560 


Gly Ser Pro Ile Gly Val Arg Leu Leu Gln Ala Leu Leu Pro Ser Ala 
                565                 570                 575     


Ser Gln Pro Gln Pro His His His His His His 
            580                 585         


<210>  3
<211>  364
<212>  PRT
<213>  artificial

<220>
<223>  activated Kumamolisin 1, backbone variant

<400>  3

Ala Ala Pro Thr Ala Tyr Thr Pro Leu Asp Val Ala Gln Ala Tyr Gln 
1               5                   10                  15      


Phe Pro Glu Gly Leu Asp Gly Gln Gly Gln Cys Ile Ala Ile Ile Glu 
            20                  25                  30          


Leu Gly Gly Gly Tyr Asp Glu Ala Ser Leu Ala Gln Tyr Phe Ala Ser 
        35                  40                  45              


Leu Gly Val Pro Ala Pro Gln Val Val Ser Val Ser Val Asp Gly Ala 
    50                  55                  60                  


Ser Asn Gln Pro Thr Gly Asp Pro Ser Gly Pro Asp Gly Glu Val Glu 
65                  70                  75                  80  


Leu Asp Ile Glu Val Ala Gly Ala Leu Ala Pro Gly Ala Lys Phe Ala 
                85                  90                  95      


Val Tyr Phe Ala Pro Asn Thr Asp Ala Gly Phe Leu Asp Ala Ile Thr 
            100                 105                 110         


Thr Ala Ile His Asp Pro Thr Leu Lys Pro Ser Val Val Ser Ile Ser 
        115                 120                 125             


Trp Gly Gly Pro Glu Asp Ser Trp Thr Ser Ala Ala Ile Ala Ala Met 
    130                 135                 140                 


Asn Arg Ala Phe Leu Asp Ala Ala Ala Leu Gly Val Thr Val Leu Ala 
145                 150                 155                 160 


Ala Ala Gly Asp Ser Gly Ser Thr Asp Gly Glu Gln Asp Gly Leu Tyr 
                165                 170                 175     


His Val Asp Phe Pro Ala Ala Ser Pro Tyr Val Leu Ala Cys Gly Gly 
            180                 185                 190         


Thr Arg Leu Val Ala Ser Gly Gly Arg Ile Ala Gln Glu Thr Val Trp 
        195                 200                 205             


Asn Asp Gly Pro Asp Gly Gly Ala Thr Gly Gly Gly Val Ser Arg Ile 
    210                 215                 220                 


Phe Pro Leu Pro Ala Trp Gln Glu His Ala Asn Val Pro Pro Ser Ala 
225                 230                 235                 240 


Asn Pro Gly Ala Ser Ser Gly Arg Gly Val Pro Asp Leu Ala Gly Asn 
                245                 250                 255     


Ala Asp Pro Ala Thr Gly Tyr Glu Val Val Ile Asp Gly Glu Ala Thr 
            260                 265                 270         


Val Ile Gly Gly Thr Ser Ala Val Ala Pro Leu Phe Ala Ala Leu Val 
        275                 280                 285             


Ala Arg Ile Asn Gln Lys Leu Gly Lys Ala Val Gly Tyr Leu Asn Pro 
    290                 295                 300                 


Thr Leu Tyr Gln Leu Pro Ala Asp Val Phe His Asp Ile Thr Glu Gly 
305                 310                 315                 320 


Asn Asn Asp Ile Ala Asn Arg Ala Gln Ile Tyr Gln Ala Gly Pro Gly 
                325                 330                 335     


Trp Asp Pro Cys Thr Gly Leu Gly Ser Pro Ile Gly Val Arg Leu Leu 
            340                 345                 350         


Gln Ala Leu Leu Pro Ser Ala Ser Gln Pro Gln Pro 
        355                 360                 


<210>  4
<211>  553
<212>  PRT
<213>  Alicyclobacillus sendaiensis

<400>  4

Met Ser Asp Met Glu Lys Pro Trp Lys Glu Gly Glu Glu Ala Arg Ala 
1               5                   10                  15      


Val Leu Gln Gly His Ala Arg Ala Gln Ala Pro Gln Ala Val Asp Lys 
            20                  25                  30          


Gly Pro Val Ala Gly Asp Glu Arg Met Ala Val Thr Val Val Leu Arg 
        35                  40                  45              


Arg Gln Arg Ala Gly Glu Leu Ala Ala His Val Glu Arg Gln Ala Ala 
    50                  55                  60                  


Ile Ala Pro His Ala Arg Glu His Leu Lys Arg Glu Ala Phe Ala Ala 
65                  70                  75                  80  


Ser His Gly Ala Ser Leu Asp Asp Phe Ala Glu Leu Arg Arg Phe Ala 
                85                  90                  95      


Asp Ala His Gly Leu Ala Leu Asp Arg Ala Asn Val Ala Ala Gly Thr 
            100                 105                 110         


Ala Val Leu Ser Gly Pro Val Asp Ala Ile Asn Arg Ala Phe Gly Val 
        115                 120                 125             


Glu Leu Arg His Phe Asp His Pro Asp Gly Ser Tyr Arg Ser Tyr Leu 
    130                 135                 140                 


Gly Glu Val Thr Val Pro Ala Ser Ile Ala Pro Met Ile Glu Ala Val 
145                 150                 155                 160 


Leu Gly Leu Asp Thr Arg Pro Val Ala Arg Pro His Phe Arg Met Gln 
                165                 170                 175     


Arg Arg Ala Glu Gly Gly Phe Glu Ala Arg Ser Gln Ala Ala Ala Pro 
            180                 185                 190         


Thr Ala Tyr Thr Pro Leu Asp Val Ala Gln Ala Tyr Gln Phe Pro Glu 
        195                 200                 205             


Gly Leu Asp Gly Gln Gly Gln Cys Ile Ala Ile Ile Glu Leu Gly Gly 
    210                 215                 220                 


Gly Tyr Asp Glu Ala Ser Leu Ala Gln Tyr Phe Ala Ser Leu Gly Val 
225                 230                 235                 240 


Pro Ala Pro Gln Val Val Ser Val Ser Val Asp Gly Ala Ser Asn Gln 
                245                 250                 255     


Pro Thr Gly Asp Pro Ser Gly Pro Asp Gly Glu Val Glu Leu Asp Ile 
            260                 265                 270         


Glu Val Ala Gly Ala Leu Ala Pro Gly Ala Lys Phe Ala Val Tyr Phe 
        275                 280                 285             


Ala Pro Asn Thr Asp Ala Gly Phe Leu Asp Ala Ile Thr Thr Ala Ile 
    290                 295                 300                 


His Asp Pro Thr Leu Lys Pro Ser Val Val Ser Ile Ser Trp Gly Gly 
305                 310                 315                 320 


Pro Glu Asp Ser Trp Thr Ser Ala Ala Ile Ala Ala Met Asn Arg Ala 
                325                 330                 335     


Phe Leu Asp Ala Ala Ala Leu Gly Val Thr Val Leu Ala Ala Ala Gly 
            340                 345                 350         


Asp Ser Gly Ser Thr Asp Gly Glu Gln Asp Gly Leu Tyr His Val Asp 
        355                 360                 365             


Phe Pro Ala Ala Ser Pro Tyr Val Leu Ala Cys Gly Gly Thr Arg Leu 
    370                 375                 380                 


Val Ala Ser Gly Gly Arg Ile Ala Gln Glu Thr Val Trp Asn Asp Gly 
385                 390                 395                 400 


Pro Asp Gly Gly Ala Thr Gly Gly Gly Val Ser Arg Ile Phe Pro Leu 
                405                 410                 415     


Pro Ala Trp Gln Glu His Ala Asn Val Pro Pro Ser Ala Asn Pro Gly 
            420                 425                 430         


Ala Ser Ser Gly Arg Gly Val Pro Asp Leu Ala Gly Asn Ala Asp Pro 
        435                 440                 445             


Ala Thr Gly Tyr Glu Val Val Ile Asp Gly Glu Ala Thr Val Ile Gly 
    450                 455                 460                 


Gly Thr Ser Ala Val Ala Pro Leu Phe Ala Ala Leu Val Ala Arg Ile 
465                 470                 475                 480 


Asn Gln Lys Leu Gly Lys Ala Val Gly Tyr Leu Asn Pro Thr Leu Tyr 
                485                 490                 495     


Gln Leu Pro Ala Asp Val Phe His Asp Ile Thr Glu Gly Asn Asn Asp 
            500                 505                 510         


Ile Ala Asn Arg Ala Gln Ile Tyr Gln Ala Gly Pro Gly Trp Asp Pro 
        515                 520                 525             


Cys Thr Gly Leu Gly Ser Pro Ile Gly Val Arg Leu Leu Gln Ala Leu 
    530                 535                 540                 


Leu Pro Ser Ala Ser Gln Pro Gln Pro 
545                 550             


<210>  5
<211>  29
<212>  PRT
<213>  artificial

<220>
<223>  sacB signal peptide

<400>  5

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


Thr Ala Leu Leu Ala Gly Gly Ala Thr Gln Ala Phe Ala 
            20                  25                  


