               SEQUENCE LISTING

<110> Global Bioenergies

<120> 3-Methylcrotonic acid decarboxylase (MDC) variants

<130> AE1573 PCT S3

<150> EP 21 166 368.7
<151> 2021-03-31

<160> 9

<170> BiSSAP 1.3.6

<210> 1
<211> 490
<212> PRT
<213> Yersinia frederiksenii


<400> 1
Met Phe Leu Lys Ser Leu Arg Glu Gln Ile Asn Leu Leu Lys Arg Gln 
1               5                   10                  15      
Gly Asp Leu Arg Ile Ile Asn Arg Pro Val Asp Ser Tyr Leu Glu Ala 
            20                  25                  30          
Ala Ala Ile Ile Arg Arg Ser Thr Glu Thr Lys Ala Pro Val Pro Leu 
        35                  40                  45              
Met Thr Asn Ile Lys Gly Tyr Pro Gly Cys Ser Ile Val Gly Gly Leu 
    50                  55                  60                  
Ala Ala Leu Ser Ser Asn Pro Glu Tyr Pro Leu Ser Arg Val Ala Met 
65                  70                  75                  80  
Ser Leu Gly Leu Ser Thr Thr Ala Ser Ala Gln Ser Ile Val Gln Tyr 
                85                  90                  95      
Leu Ile Asp Gly Leu Lys Lys Thr Pro Tyr Leu Pro Val Glu Ile Asp 
            100                 105                 110         
Ser Asn Ser Ala Pro Cys Lys Gln Asn Ile Leu Arg Gly Glu Asp Ala 
        115                 120                 125             
Ser Leu Ala His Phe Pro Ile Pro Gln Val His Gln His Asp Gly Asn 
    130                 135                 140                 
Arg Tyr Val Asn Thr Trp Gly Val Phe Ile Val Glu Ser Pro Asp Gly 
145                 150                 155                 160 
Gln Trp Cys Asn Trp Ser Ile Gln Arg Val Gln Tyr Leu Asn Asp Arg 
                165                 170                 175     
Gln Met Ile Ala Leu Val Phe Pro Ser Gln His Ile Ser Asp Ile Trp 
            180                 185                 190         
Glu Glu Trp Val Lys Ile Gly Lys Pro Met Pro Tyr Ala Leu Val Gln 
        195                 200                 205             
Gly Cys Glu Pro Ile Val Pro Tyr Ile Ala Gly Leu Pro Leu Leu Glu 
    210                 215                 220                 
Arg Asn Val Asn Glu Ala Glu Tyr Ile Gly Ala Leu Tyr Gly Lys Gly 
225                 230                 235                 240 
Ile Glu Val Val Lys Cys Glu Thr His Asp Leu Arg Val Pro Ala Ser 
                245                 250                 255     
Ser Glu Val Val Ile Glu Gly Tyr Phe Ala Ile Thr Arg Glu Gly Val 
            260                 265                 270         
Glu Gly Pro Phe Gly Glu Phe Ala Gly Tyr Ile Pro Asn Glu Asn Ser 
        275                 280                 285             
Leu Gln Pro Ile Leu Ser Ile Thr Ala Ile Ser Trp Gln Asp Asp Pro 
    290                 295                 300                 
Ile Trp Pro Val Val Ala Glu Gly Lys Pro Val Asp Glu Tyr His Thr 
305                 310                 315                 320 
Cys Ser Gly Ile Gly Asp Ala Ala Ala Ile Gln Asn Val Leu Leu Thr 
                325                 330                 335     
Ala Asn Leu Pro Val Ser Phe Val Trp Val Pro Leu Tyr Ser Ala Cys 
            340                 345                 350         
His Leu Met Ile Val Ser Ile Asn Arg His Trp Arg Gln Thr His Pro 
        355                 360                 365             
Asp Leu Thr Ser Glu Gln Leu Thr Lys Lys Val Gly Asp Val Ile His 
    370                 375                 380                 
Gln Thr Arg Asn Ser Ile Lys Leu Pro Lys Ile Val Val Leu Asp Asp 
385                 390                 395                 400 
Asp Val Asp Pro Ala Asp Thr Glu Ser Leu Leu Trp Ala Leu Ala Thr 
                405                 410                 415     
Arg Ile His Pro Glu Arg Lys Arg Tyr Phe Tyr Glu Ser Cys Ile Leu 
            420                 425                 430         
Pro Leu Leu Ser Cys Tyr Ser Glu Glu Glu Arg His Asn Arg Lys Gly 
        435                 440                 445             
Lys Arg Val Ile Ile Asp Ala Leu Leu Pro Glu Asn His Gly Ala Ile 
    450                 455                 460                 
Ser Ser Phe Asp Tyr Ala Tyr Pro Asp Asp Ile Lys Gln Arg Val Leu 
465                 470                 475                 480 
Met His Trp Glu Thr Asp Phe Ser His Asp 
                485                 490 

<210> 2
<211> 189
<212> PRT
<213> Escherichia coli K-12


<400> 2
Met Lys Arg Leu Ile Val Gly Ile Ser Gly Ala Ser Gly Ala Ile Tyr 
1               5                   10                  15      
Gly Val Arg Leu Leu Gln Val Leu Arg Asp Val Thr Asp Ile Glu Thr 
            20                  25                  30          
His Leu Val Met Ser Gln Ala Ala Arg Gln Thr Leu Ser Leu Glu Thr 
        35                  40                  45              
Asp Phe Ser Leu Arg Glu Val Gln Ala Leu Ala Asp Val Thr His Asp 
    50                  55                  60                  
Ala Arg Asp Ile Ala Ala Ser Ile Ser Ser Gly Ser Phe Gln Thr Leu 
65                  70                  75                  80  
Gly Met Val Ile Leu Pro Cys Ser Ile Lys Thr Leu Ser Gly Ile Val 
                85                  90                  95      
His Ser Tyr Thr Asp Gly Leu Leu Thr Arg Ala Ala Asp Val Val Leu 
            100                 105                 110         
Lys Glu Arg Arg Pro Leu Val Leu Cys Val Arg Glu Thr Pro Leu His 
        115                 120                 125             
Leu Gly His Leu Arg Leu Met Thr Gln Ala Ala Glu Ile Gly Ala Val 
    130                 135                 140                 
Ile Met Pro Pro Val Pro Ala Phe Tyr His Arg Pro Gln Ser Leu Asp 
145                 150                 155                 160 
Asp Val Ile Asn Gln Thr Val Asn Arg Val Leu Asp Gln Phe Ala Ile 
                165                 170                 175     
Thr Leu Pro Glu Asp Leu Phe Ala Arg Trp Gln Gly Ala 
            180                 185                 

<210> 3
<211> 229
<212> PRT
<213> Candida albicans


<220> 
<223> Candida albicans (strain SC5314 / ATCC MYA-2876)

<400> 3
Met Ile Ala Arg Val Cys Leu Arg Arg Ser Asn Val Leu Pro Ile Phe 
1               5                   10                  15      
Gln Ile Pro Ser Arg Lys Tyr Ser Ile Asn Tyr Glu Lys Val Asn Asn 
            20                  25                  30          
Ser Ile Tyr Asn Asn Val Ile Lys Pro Lys Arg Ile Val Leu Ala Ile 
        35                  40                  45              
Thr Gly Ala Thr Gly Thr Gln Ile Gly Val Arg Leu Leu Glu Ile Leu 
    50                  55                  60                  
Lys Glu Leu Gly Val Glu Thr His Leu Val Met Ser Lys Trp Gly Ile 
65                  70                  75                  80  
Ala Thr Leu Lys Tyr Glu Thr Asp Tyr Gln Val Asp Tyr Val Thr Ser 
                85                  90                  95      
Leu Ala Thr Lys Thr Tyr Ser Ala Arg Asp Val Thr Ala Pro Ile Ser 
            100                 105                 110         
Ser Gly Ser Phe Val His Asp Gly Met Ile Val Ala Pro Cys Ser Met 
        115                 120                 125             
Lys Ser Leu Ser Ala Ile Arg Thr Gly Phe Thr Glu Asp Leu Ile Val 
    130                 135                 140                 
Arg Ala Ala Asp Val Ser Leu Lys Glu Arg Arg Lys Leu Leu Leu Val 
145                 150                 155                 160 
Ala Arg Glu Thr Pro Leu Ser Asp Ile His Leu Asp Asn Met Leu Tyr 
                165                 170                 175     
Leu Ser Arg Met Gly Val Thr Ile Phe Pro Pro Val Pro Ala Phe Tyr 
            180                 185                 190         
Thr Lys Pro Lys Thr Ile Asp Asp Ile Val Glu Gln Thr Cys Gly Arg 
        195                 200                 205             
Ile Leu Asp Asn Phe Gly Ile Asn Ile Asp Thr Phe Glu Arg Trp Asp 
    210                 215                 220                 
Gly Ile Asn His Arg 
225                 

<210> 4
<211> 227
<212> PRT
<213> Aspergillus niger


<400> 4
Met Phe Asn Ser Leu Leu Ser Gly Thr Thr Thr Pro Asn Ser Gly Arg 
1               5                   10                  15      
Ala Ser Pro Pro Ala Ser Glu Met Pro Ile Asp Asn Asp His Val Ala 
            20                  25                  30          
Val Ala Arg Pro Ala Pro Arg Arg Arg Arg Ile Val Val Ala Met Thr 
        35                  40                  45              
Gly Ala Thr Gly Ala Met Leu Gly Ile Lys Val Leu Ile Ala Leu Arg 
    50                  55                  60                  
Arg Leu Asn Val Glu Thr His Leu Val Met Ser Lys Trp Ala Glu Ala 
65                  70                  75                  80  
Thr Ile Lys Tyr Glu Thr Asp Tyr His Pro Ser Asn Val Arg Ala Leu 
                85                  90                  95      
Ala Asp Tyr Val His Asn Ile Asn Asp Met Ala Ala Pro Val Ser Ser 
            100                 105                 110         
Gly Ser Phe Arg Ala Asp Gly Met Ile Val Val Pro Cys Ser Met Lys 
        115                 120                 125             
Thr Leu Ala Ala Ile His Ser Gly Phe Cys Asp Asp Leu Ile Ser Arg 
    130                 135                 140                 
Thr Ala Asp Val Met Leu Lys Glu Arg Arg Arg Leu Val Leu Val Ala 
145                 150                 155                 160 
Arg Glu Thr Pro Leu Ser Glu Ile His Leu Arg Asn Met Leu Glu Val 
                165                 170                 175     
Thr Arg Ala Gly Ala Val Ile Phe Pro Pro Val Pro Ala Phe Tyr Ile 
            180                 185                 190         
Lys Ala Gly Ser Ile Glu Asp Leu Ile Asp Gln Ser Val Gly Arg Met 
        195                 200                 205             
Leu Asp Leu Phe Asp Leu Asp Thr Gly Asp Phe Glu Arg Trp Asn Gly 
    210                 215                 220                 
Trp Glu Lys 
225         

<210> 5
<211> 242
<212> PRT
<213> Saccharomyces cerevisiae S288c


<400> 5
Met Leu Leu Phe Pro Arg Arg Thr Asn Ile Ala Phe Phe Lys Thr Thr 
1               5                   10                  15      
Gly Ile Phe Ala Asn Phe Pro Leu Leu Gly Arg Thr Ile Thr Thr Ser 
            20                  25                  30          
Pro Ser Phe Leu Thr His Lys Leu Ser Lys Glu Val Thr Arg Ala Ser 
        35                  40                  45              
Thr Ser Pro Pro Arg Pro Lys Arg Ile Val Val Ala Ile Thr Gly Ala 
    50                  55                  60                  
Thr Gly Val Ala Leu Gly Ile Arg Leu Leu Gln Val Leu Lys Glu Leu 
65                  70                  75                  80  
Ser Val Glu Thr His Leu Val Ile Ser Lys Trp Gly Ala Ala Thr Met 
                85                  90                  95      
Lys Tyr Glu Thr Asp Trp Glu Pro His Asp Val Ala Ala Leu Ala Thr 
            100                 105                 110         
Lys Thr Tyr Ser Val Arg Asp Val Ser Ala Cys Ile Ser Ser Gly Ser 
        115                 120                 125             
Phe Gln His Asp Gly Met Ile Val Val Pro Cys Ser Met Lys Ser Leu 
    130                 135                 140                 
Ala Ala Ile Arg Ile Gly Phe Thr Glu Asp Leu Ile Thr Arg Ala Ala 
145                 150                 155                 160 
Asp Val Ser Ile Lys Glu Asn Arg Lys Leu Leu Leu Val Thr Arg Glu 
                165                 170                 175     
Thr Pro Leu Ser Ser Ile His Leu Glu Asn Met Leu Ser Leu Cys Arg 
            180                 185                 190         
Ala Gly Val Ile Ile Phe Pro Pro Val Pro Ala Phe Tyr Thr Arg Pro 
        195                 200                 205             
Lys Ser Leu His Asp Leu Leu Glu Gln Ser Val Gly Arg Ile Leu Asp 
    210                 215                 220                 
Cys Phe Gly Ile His Ala Asp Thr Phe Pro Arg Trp Glu Gly Ile Lys 
225                 230                 235                 240 
Ser Lys 
        

<210> 6
<211> 180
<212> PRT
<213> Cryptococcus gattii WM276


<220> 
<223> serotype B

<400> 6
Met Arg Arg Lys Arg Tyr Val Val Ala Val Thr Gly Ala Thr Gly Ala 
1               5                   10                  15      
Thr Leu Ala Ile Arg Leu Leu Gln Ala Leu Arg Ala Leu Asp Ile Glu 
            20                  25                  30          
Thr His Leu Ile Ile Ser Lys Trp Ala Val Lys Thr Leu Lys Tyr Glu 
        35                  40                  45              
Thr Asp Met Ile Glu Arg Glu Leu Lys Asp Leu Ala Asp Tyr Ser Tyr 
    50                  55                  60                  
Ser Asn Ser Asp Leu Ala Ala Pro Pro Ser Ser Gly Ser Phe Ile His 
65                  70                  75                  80  
Asp Gly Met Phe Ile Ile Pro Cys Ser Met Lys Thr Leu Ala Ala Val 
                85                  90                  95      
Arg Ile Gly Leu Gly Asp Glu Leu Ile Ser Arg Ser Ala Asp Val Cys 
            100                 105                 110         
Leu Lys Glu Gly Arg Lys Leu Met Leu Val Val Arg Glu Thr Pro Leu 
        115                 120                 125             
Asn Asp Ile His Leu Glu Asn Met Leu Phe Leu Arg Arg Ala Gly Ala 
    130                 135                 140                 
Ile Ile Phe Pro Pro Val Pro Ala Tyr Tyr Ile Arg Pro Gln Thr Ile 
145                 150                 155                 160 
Asp Asp Leu Thr Asn Gln Thr Val Gly Arg Ile Leu Asp Ser Ser Lys 
                165                 170                 175     
Cys Ser Gln Lys 
            180 

<210> 7
<211> 192
<212> PRT
<213> Bacillus subtilis


<400> 7
Met Lys Leu Val Ile Gly Met Thr Gly Ala Thr Gly Ala Ile Phe Gly 
1               5                   10                  15      
Ile Arg Leu Leu Glu Tyr Leu Lys Ala Ala Glu Ile Glu Thr His Leu 
            20                  25                  30          
Val Val Ser Pro Trp Ala Asn Val Thr Ile Thr His Glu Thr Asp Tyr 
        35                  40                  45              
Thr Leu Lys Asp Val Glu Lys Leu Ala Ser Tyr Thr Tyr Ser His Lys 
    50                  55                  60                  
Asp Gln Ala Ala Ala Ile Ser Ser Gly Ser Phe Glu Thr Asp Gly Met 
65                  70                  75                  80  
Ile Ile Ala Pro Cys Ser Met Lys Ser Leu Ala Ser Ile Arg Thr Gly 
                85                  90                  95      
Met Ala Asp Asn Leu Leu Thr Arg Ala Ala Asp Val Ile Leu Lys Glu 
            100                 105                 110         
Arg Lys Lys Leu Val Leu Leu Thr Arg Glu Thr Pro Leu Ser Gln Ile 
        115                 120                 125             
His Leu Glu Asn Met Leu Ala Leu Thr Lys Met Gly Ser Val Ile Leu 
    130                 135                 140                 
Pro Pro Met Pro Ala Phe Tyr Asn Lys Pro Ala Asp Met Asp Glu Leu 
145                 150                 155                 160 
Ile Asp His Ile Val Phe Arg Thr Leu Asp Gln Phe Gly Ile Arg Leu 
                165                 170                 175     
Pro Glu Ala Lys Arg Trp Tyr Gly Ile Glu Lys Gln Lys Gly Gly Ile 
            180                 185                 190         



<210> 8
<211> 209
<212> PRT
<213> Pseudomonas aeruginosa


<400> 8
Met Ser Gly Pro Glu Arg Ile Thr Leu Ala Met Thr Gly Ala Ser Gly 
1               5                   10                  15      
Ala Gln Tyr Gly Leu Arg Leu Leu Asp Cys Leu Val Gln Glu Glu Arg 
            20                  25                  30          
Glu Val His Phe Leu Ile Ser Lys Ala Ala Gln Leu Val Met Ala Thr 
        35                  40                  45              
Glu Thr Asp Val Ala Leu Pro Ala Lys Pro Gln Ala Met Gln Ala Phe 
    50                  55                  60                  
Leu Thr Glu Tyr Cys Gly Ala Ala Ala Gly Gln Ile Arg Val Phe Gly 
65                  70                  75                  80  
Gln Asn Asp Trp Met Ala Pro Pro Ala Ser Gly Ser Ser Ala Pro Asn 
                85                  90                  95      
Ala Met Val Ile Cys Pro Cys Ser Thr Gly Thr Leu Ser Ala Val Ala 
            100                 105                 110         
Thr Gly Ala Cys Asn Asn Leu Ile Glu Arg Ala Ala Asp Val Ala Leu 
        115                 120                 125             
Lys Glu Arg Arg Pro Leu Val Leu Val Pro Arg Glu Ala Pro Phe Ser 
    130                 135                 140                 
Ser Ile His Leu Glu Asn Met Leu Lys Leu Ser Asn Leu Gly Ala Val 
145                 150                 155                 160 
Ile Leu Pro Ala Ala Pro Gly Phe Tyr His Gln Pro Gln Ser Val Glu 
                165                 170                 175     
Asp Leu Val Asp Phe Val Val Ala Arg Ile Leu Asn Thr Leu Gly Ile 
            180                 185                 190         
Pro Gln Asp Met Leu Pro Arg Trp Gly Glu Gln His Leu Val Ser Asp 
        195                 200                 205             
Glu 
    

<210> 9
<211> 219
<212> PRT
<213> Enterobacter sp.


<220> 
<223> Enterobacter sp. DC4

<400> 9
Met Leu Arg Gln Val Arg Ala Asn Ala Leu Thr Cys Asn Ser Pro Gln 
1               5                   10                  15      
Asn Pro Ala Gln Ser Ala Leu Lys Ser Val Arg Ala Lys Ile Met Lys 
            20                  25                  30          
Arg Leu Ile Val Gly Leu Ser Gly Ala Ser Gly Ala Ile Tyr Gly Val 
        35                  40                  45              
Arg Leu Leu Gln Val Leu Arg Asn Val Ala Glu Val Glu Thr His Leu 
    50                  55                  60                  
Val Met Ser Gln Ala Ala Arg Gln Thr Leu Ser Leu Glu Thr Asp Leu 
65                  70                  75                  80  
Ser Leu Arg Asp Val Gln Ala Leu Ala Asp Val Val His Asp Ala Arg 
                85                  90                  95      
Asp Ile Ala Ala Ser Ile Ser Ser Gly Ser Phe Lys Thr Ala Gly Met 
            100                 105                 110         
Val Ile Leu Pro Cys Ser Ile Lys Thr Leu Ser Gly Ile Val Asn Ser 
        115                 120                 125             
Tyr Thr Asp Thr Leu Val Thr Arg Ala Ala Asp Val Val Leu Lys Glu 
    130                 135                 140                 
Arg Arg Pro Leu Val Leu Cys Val Arg Glu Thr Pro Leu His Leu Gly 
145                 150                 155                 160 
His Leu Arg Leu Met Thr Gln Ala Ala Glu Leu Gly Ala Ile Ile Met 
                165                 170                 175     
Pro Pro Val Pro Ala Phe Tyr His Arg Pro Thr Ser Leu Asp Asp Val 
            180                 185                 190         
Ile Asn Gln Thr Val Asn Arg Val Leu Asp Gln Phe Asp Ile Asp Leu 
        195                 200                 205             
Pro Glu Asp Leu Phe Thr Arg Trp Gln Gly Ala 
    210                 215                 

