                         SEQUENCE LISTING

<110>  Roche Diabetes Care GmbH et al.
       
 
<120>  Mutant 3 Hydroxybutyrate Dehygrogenase from Alcaligenes faecalis 
       as well as methods and uses involving the same

<130>  R69992PC

<160>  16    

<170>  PatentIn version 3.5

<210>  1
<211>  260
<212>  PRT
<213>  Alcaligenes faecalis

<400>  1

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Ile Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Ala Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Ala Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Ala His 
    130                 135                 140                 


Gly Leu Val Ala Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Gly Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Trp Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


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


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Leu Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  2
<211>  192
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  extended core sequence of 3-HBDH from Alcaligenes faecalis

<400>  2

Ala Asp Leu Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala 
1               5                   10                  15      


Ala Glu Ala Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile 
            20                  25                  30          


Gln His Thr Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala 
        35                  40                  45              


Ile Ile Ala Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Ala 
    50                  55                  60                  


Leu Pro Ile Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala 
65                  70                  75                  80  


Ser Ala His Gly Leu Val Ala Ser Val Asn Lys Ser Ala Tyr Val Ala 
                85                  90                  95      


Ala Lys His Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn 
            100                 105                 110         


Ala Gly Lys Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Trp Val Arg 
        115                 120                 125             


Thr Pro Leu Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly 
    130                 135                 140                 


Ile Asp Ile Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro 
145                 150                 155                 160 


Ser Leu Gln Phe Val Thr Pro Glu Gln Leu Gly Gly Ala Ala Val Phe 
                165                 170                 175     


Leu Ser Ser Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Leu 
            180                 185                 190         


<210>  3
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  3

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Ile Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Ala Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Ala Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Ala His 
    130                 135                 140                 


Gly Leu Val Ala Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Gly Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Trp Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


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


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  4
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  4

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Ile Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Ala Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Ala Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Ala His 
    130                 135                 140                 


Gly Leu Val Ala Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Gly Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Trp Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  5
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  5

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Ile Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Ala Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Ala His 
    130                 135                 140                 


Gly Leu Val Ala Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Gly Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Trp Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  6
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  6

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Ile Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Ala Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Ala His 
    130                 135                 140                 


Gly Leu Arg Ala Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Gly Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Trp Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  7
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  7

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Ile Asn Gly Phe Gly Gln Gly Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Arg Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Ala Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Val His 
    130                 135                 140                 


Gly Leu Val Gly Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Gly Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Trp Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  8
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  8

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Ile Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Ala Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Val His 
    130                 135                 140                 


Gly Leu Val Gly Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Gly Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Trp Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  9
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  9

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Leu Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Gly Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Val His 
    130                 135                 140                 


Gly Leu Val Gly Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Val Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Phe Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  10
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  10

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Ile Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Gly Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Val His 
    130                 135                 140                 


Gly Leu Val Gly Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Thr Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Phe Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  11
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  11

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Ile Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Ala Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Val His 
    130                 135                 140                 


Gly Leu Val Gly Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Gly Leu Glu Asn Ala Thr Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Trp Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  12
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  12

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


Gly Glu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Ile Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Gly Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Val His 
    130                 135                 140                 


Gly Leu Val Gly Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Gly Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Phe Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  13
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  13

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Leu Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Gly Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Val His 
    130                 135                 140                 


Gly Leu Val Gly Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Ile Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Phe Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  14
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  14

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Leu Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Gly Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Val His 
    130                 135                 140                 


Gly Leu Val Gly Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Gly Leu Glu Asn Ala Gly Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Phe Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  15
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  15

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Leu Asn Gly Phe Gly Gln Pro Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Gly Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Val His 
    130                 135                 140                 


Gly Leu Val Gly Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Val Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Phe Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Val Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


<210>  16
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutant 3-HBDH

<400>  16

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


Gly Leu Ala Met Ala Thr Glu Leu Ala Lys Ala Gly Ala Asp Val Val 
            20                  25                  30          


Leu Asn Gly Phe Gly Lys Gly Glu Asp Ile Glu Arg Glu Arg Ser Thr 
        35                  40                  45              


Leu Glu Ser Lys Phe Gly Val Lys Ala Tyr Tyr Leu Asn Val Asp Leu 
    50                  55                  60                  


Ser Asp Ala Gln Ala Thr Arg Asp Phe Ile Ala Lys Ala Ala Glu Ala 
65                  70                  75                  80  


Leu Gly Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Gln His Thr 
                85                  90                  95      


Ala Pro Ile Glu Glu Phe Pro Val Asp Lys Trp Asn Ala Ile Ile Ala 
            100                 105                 110         


Leu Asn Leu Ser Ala Val Phe His Gly Thr Ala Ala Gly Leu Pro Ile 
        115                 120                 125             


Met Gln Lys Gln Gly Trp Gly Arg Ile Ile Asn Ile Ala Ser Val His 
    130                 135                 140                 


Gly Leu Val Gly Ser Val Asn Lys Ser Ala Tyr Val Ala Ala Lys His 
145                 150                 155                 160 


Gly Val Val Gly Leu Thr Lys Val Thr Ala Leu Glu Asn Ala Val Lys 
                165                 170                 175     


Gly Ile Thr Cys Asn Ala Ile Cys Pro Gly Phe Val Arg Thr Pro Leu 
            180                 185                 190         


Val Glu Lys Gln Ile Glu Ala Ile Ser Gln Gln Lys Gly Ile Asp Ile 
        195                 200                 205             


Glu Ala Ala Ala Arg Glu Leu Leu Ala Glu Lys Gln Pro Ser Leu Gln 
    210                 215                 220                 


Phe Val Thr Pro Glu Gln Leu Gly Lys Thr Ala Val Phe Leu Ser Ser 
225                 230                 235                 240 


Ala Ala Ala Asp Gln Met Thr Gly Thr Thr Leu Ser Ile Asp Gly Gly 
                245                 250                 255     


Trp Thr Ala Arg 
            260 


