                         SEQUENCE LISTING

<110>  VIB VZW
       KATHOLIEKE UNIVERSITEIT LEUVEN, K.U.LEUVEN R&D
 
<120>  MEANS AND METHODS TO IMPROVE FERMENTATION EFFICIENCY

<130>  JoTh/AST2/692

<150>  EP 20179988.9
<151>  2020-06-15

<160>  6     

<170>  PatentIn version 3.5

<210>  1
<211>  430
<212>  PRT
<213>  Saccharomyces cerevisiae

<400>  1

Met Ala Glu Lys Ile Leu Glu Asn Lys Asp Pro Lys Leu Glu Ala Met 
1               5                   10                  15      


Thr Val Asp His Glu Val Ser Ala Pro Lys Pro Ile Pro Val Asp Glu 
            20                  25                  30          


Pro Thr Leu Thr Arg Val Ala Arg Pro Leu Arg His Val Arg His Ile 
        35                  40                  45              


Pro Val Lys Ser Leu Val Phe His Ser Lys His Gly Pro Ile Thr Phe 
    50                  55                  60                  


Ser Tyr Glu Asn Lys Ile Lys Leu Pro Ile Ser Lys Asn Lys Leu Val 
65                  70                  75                  80  


Val Gln Val Asn Tyr Val Gly Leu Asn Pro Val Asp Met Lys Ile Arg 
                85                  90                  95      


Asn Gly Tyr Thr Lys Pro Ile Tyr Gly Glu Ala Gly Ile Gly Arg Glu 
            100                 105                 110         


Tyr Ser Gly Val Ile Thr His Val Gly Asp Asn Leu Thr Asn Arg Trp 
        115                 120                 125             


Asn Val Gly Asp Asp Val Tyr Gly Ile Tyr Tyr His Pro Lys Leu Ala 
    130                 135                 140                 


Ile Gly Ala Leu Gln Ser Ser Leu Leu Ile Asp Pro Arg Val Asp Pro 
145                 150                 155                 160 


Ile Leu Met Arg Pro Lys Asn Thr Leu Ser Pro Glu Lys Ala Ala Gly 
                165                 170                 175     


Ser Leu Phe Cys Leu Gly Thr Ala Leu Asn Leu Leu Ala Gln Leu Lys 
            180                 185                 190         


Glu Lys Asp Gln Leu Asn Thr Glu Ser Asn Val Leu Ile Asn Gly Gly 
        195                 200                 205             


Thr Ser Ser Val Gly Met Phe Ala Ile Gln Leu Leu Lys Arg Tyr Tyr 
    210                 215                 220                 


Lys Val Ser Lys Lys Leu Val Val Val Thr Ser Gly Asn Gly Ala Ala 
225                 230                 235                 240 


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


Asn Tyr Leu Ser Cys Arg Gly Lys Ser Ser Lys Pro Leu Arg Arg Met 
            260                 265                 270         


Leu Asp Thr Gly Lys Val Val Asp Tyr Asp Asp Phe Asn Thr Leu Lys 
        275                 280                 285             


Glu Thr Glu Asp Tyr Thr Gln Gly Lys Phe Asn Val Val Leu Asp Phe 
    290                 295                 300                 


Ile Gly Gly Tyr Asp Ile Leu Ser His Ser Ser Ser Leu Ile His Ala 
305                 310                 315                 320 


Lys Gly Ala Tyr Ile Thr Thr Val Gly Asp Tyr Val Gly Asn Tyr Lys 
                325                 330                 335     


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


Met Phe Gly Ser Met Leu Trp Ser Tyr Asp Tyr Ser His Phe Tyr Phe 
        355                 360                 365             


Asp Pro Asn Ile Lys Ile Ile Pro Lys Lys Asn Asp Trp Ile His Glu 
    370                 375                 380                 


Cys Gly Lys Leu Leu Asn Glu Gly Val Val Asp Cys Val Val Asp Lys 
385                 390                 395                 400 


Val Tyr Ser Trp Lys Asn Phe Lys Glu Ala Phe Ser Tyr Met Ala Thr 
                405                 410                 415     


Gln Arg Ala Gln Gly Lys Leu Ile Met Lys Val Glu Gly Phe 
            420                 425                 430 


<210>  2
<211>  430
<212>  PRT
<213>  Saccharomyces cerevisiae

<400>  2

Met Ala Glu Lys Ile Leu Glu Asn Lys Asp Pro Lys Leu Glu Ala Met 
1               5                   10                  15      


Thr Val Asp His Glu Val Ser Ala Pro Lys Pro Ile Pro Val Asp Glu 
            20                  25                  30          


Pro Thr Leu Thr Arg Val Ala Arg Pro Leu Arg His Val Arg His Ile 
        35                  40                  45              


Pro Val Lys Ser Leu Val Phe His Ser Lys His Gly Pro Ile Thr Phe 
    50                  55                  60                  


Ser Tyr Glu Asn Lys Ile Lys Leu Pro Ile Ser Lys Asn Lys Leu Val 
65                  70                  75                  80  


Val Gln Val Asn Tyr Val Gly Leu Asn Pro Val Asp Met Lys Ile Arg 
                85                  90                  95      


Asn Gly Tyr Thr Lys Pro Ile Tyr Gly Glu Ala Gly Ile Gly Arg Glu 
            100                 105                 110         


Tyr Ser Gly Val Ile Thr His Val Gly Asp Asn Leu Thr Asn Arg Trp 
        115                 120                 125             


Asn Val Gly Asp Asp Val Tyr Gly Ile Tyr Tyr His Pro Lys Leu Ala 
    130                 135                 140                 


Ile Gly Ala Leu Gln Ser Ser Leu Leu Ile Asp Pro Arg Val Asp Pro 
145                 150                 155                 160 


Ile Leu Met Arg Pro Lys Asn Thr Leu Ser Pro Glu Lys Ala Ala Gly 
                165                 170                 175     


Ser Leu Phe Cys Leu Gly Thr Ala Leu Asn Leu Leu Ala Gln Leu Lys 
            180                 185                 190         


Glu Lys Asp Gln Leu Asn Thr Glu Ser Asn Val Leu Ile Asn Gly Gly 
        195                 200                 205             


Thr Ser Ser Val Gly Met Phe Ala Ile Gln Leu Leu Lys Arg Tyr Tyr 
    210                 215                 220                 


Lys Val Ser Lys Lys Leu Val Val Val Thr Ser Gly Asn Gly Ala Ala 
225                 230                 235                 240 


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


Asn Tyr Leu Ser Cys Arg Gly Lys Ser Ser Lys Pro Leu Arg Arg Met 
            260                 265                 270         


Leu Asp Thr Gly Lys Val Val Asp Tyr Asp Asp Phe Asn Thr Leu Lys 
        275                 280                 285             


Glu Thr Glu Asp Tyr Thr Gln Gly Lys Phe Asn Val Val Leu Asp Phe 
    290                 295                 300                 


Ile Gly Gly Tyr Asp Ile Leu Ser His Ser Ser Ser Leu Ile His Ala 
305                 310                 315                 320 


Lys Gly Ala Tyr Ile Thr Thr Val Gly Asp Tyr Val Gly Asn Tyr Lys 
                325                 330                 335     


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


Met Phe Gly Ser Met Leu Trp Ser Tyr Asp Tyr Ser His Phe Tyr Phe 
        355                 360                 365             


Asp Pro Asn Ile Lys Ile Ile Pro Lys Lys Asn Asp Trp Ile His Glu 
    370                 375                 380                 


Cys Gly Lys Leu Leu Asn Glu Gly Val Val Asp Cys Val Val Asp Lys 
385                 390                 395                 400 


Val Tyr Ser Trp Lys Ile Phe Lys Glu Ala Phe Ser Tyr Met Ala Thr 
                405                 410                 415     


Gln Arg Ala Gln Gly Lys Leu Ile Met Lys Val Glu Gly Phe 
            420                 425                 430 


<210>  3
<211>  429
<212>  PRT
<213>  Saccharomyces cerevisiae

<400>  3

Met Ala Lys Asp Ile Leu Lys Asn Gln Asp Pro Lys Leu Gln Ala Met 
1               5                   10                  15      


Ile Val Glu His Ser Ala Pro Ala Pro Lys Glu Ile Pro Met Asp Ala 
            20                  25                  30          


Pro Val Leu Lys Arg Val Ala Arg Pro Leu Arg His Val Lys Phe Ile 
        35                  40                  45              


Pro Ile Lys Ser Leu Ile Phe His Thr Lys Thr Gly Pro Met Asp Phe 
    50                  55                  60                  


Ser Tyr Glu Lys Lys Ile Lys Thr Pro Ile Pro Lys Asn Lys Ile Val 
65                  70                  75                  80  


Val Arg Val Ser Asn Val Gly Leu Asn Pro Val Asp Met Lys Ile Arg 
                85                  90                  95      


Asn Gly Tyr Thr Ser Ser Ile Tyr Gly Glu Ile Gly Leu Gly Arg Glu 
            100                 105                 110         


Tyr Ser Gly Val Ile Thr Glu Val Gly Glu Asn Leu Asn Tyr Ala Trp 
        115                 120                 125             


His Val Gly Asp Glu Val Tyr Gly Ile Tyr Tyr His Pro His Leu Ala 
    130                 135                 140                 


Val Gly Cys Leu Gln Ser Ser Ile Leu Val Asp Pro Lys Val Asp Pro 
145                 150                 155                 160 


Ile Leu Leu Arg Pro Glu Ser Val Ser Ala Glu Glu Ala Ala Gly Ser 
                165                 170                 175     


Leu Phe Cys Leu Ala Thr Gly Tyr Asn Ile Leu Asn Lys Leu Ser Lys 
            180                 185                 190         


Asn Lys Tyr Leu Lys Gln Asp Ser Asn Val Leu Ile Asn Gly Gly Thr 
        195                 200                 205             


Ser Ser Val Gly Met Phe Val Ile Gln Leu Leu Lys Arg His Tyr Lys 
    210                 215                 220                 


Leu Gln Lys Lys Leu Val Ile Val Thr Ser Ala Asn Gly Pro Gln Val 
225                 230                 235                 240 


Leu Gln Glu Lys Phe Pro Asp Leu Ala Asp Glu Met Ile Phe Ile Asp 
                245                 250                 255     


Tyr Leu Thr Cys Arg Gly Lys Ser Ser Lys Pro Leu Arg Lys Met Leu 
            260                 265                 270         


Glu Glu Lys Lys Ile Ser Gln Tyr Asp Pro Val Glu Asp Lys Glu Thr 
        275                 280                 285             


Ile Leu Asn Tyr Asn Glu Gly Lys Phe Asp Val Val Leu Asp Phe Val 
    290                 295                 300                 


Gly Gly Tyr Asp Ile Leu Ser His Ser Ser Ser Leu Ile His Gly Gly 
305                 310                 315                 320 


Gly Ala Tyr Val Thr Thr Val Gly Asp Tyr Val Ala Asn Tyr Lys Glu 
                325                 330                 335     


Asp Ile Phe Asp Ser Trp Asp Asn Pro Ser Ala Asn Ala Arg Lys Met 
            340                 345                 350         


Phe Gly Ser Ile Ile Trp Ser Tyr Asn Tyr Thr His Tyr Tyr Phe Asp 
        355                 360                 365             


Pro Asn Ala Lys Thr Ala Ser Ala Asn Asn Asp Trp Ile Glu Gln Cys 
    370                 375                 380                 


Gly Asp Phe Leu Lys Asn Gly Thr Val Lys Cys Val Val Asp Lys Val 
385                 390                 395                 400 


Tyr Asp Trp Lys Asp His Lys Glu Ala Phe Ser Tyr Met Ala Thr Gln 
                405                 410                 415     


Arg Ala Gln Gly Lys Leu Ile Met Asn Val Glu Lys Phe 
            420                 425                 


<210>  4
<211>  429
<212>  PRT
<213>  Saccharomyces cerevisiae

<400>  4

Met Ala Lys Asp Ile Leu Lys Asn Gln Asp Pro Lys Leu Gln Ala Met 
1               5                   10                  15      


Ile Val Glu His Ser Ala Pro Ala Pro Lys Glu Ile Pro Met Asp Ala 
            20                  25                  30          


Pro Val Leu Lys Arg Val Ala Arg Pro Leu Arg His Val Lys Phe Ile 
        35                  40                  45              


Pro Ile Lys Ser Leu Ile Phe His Thr Lys Thr Gly Pro Met Asp Phe 
    50                  55                  60                  


Ser Tyr Glu Lys Lys Ile Lys Thr Pro Ile Pro Lys Asn Lys Ile Val 
65                  70                  75                  80  


Val Arg Val Ser Asn Val Gly Leu Asn Pro Val Asp Met Lys Ile Arg 
                85                  90                  95      


Asn Gly Tyr Thr Ser Ser Ile Tyr Gly Glu Ile Gly Leu Gly Arg Glu 
            100                 105                 110         


Tyr Ser Gly Val Ile Thr Glu Val Gly Glu Asn Leu Asn Tyr Ala Trp 
        115                 120                 125             


His Val Gly Asp Glu Val Tyr Gly Ile Tyr Tyr His Pro His Leu Ala 
    130                 135                 140                 


Val Gly Cys Leu Gln Ser Ser Ile Leu Val Asp Pro Lys Val Asp Pro 
145                 150                 155                 160 


Ile Leu Leu Arg Pro Glu Ser Val Ser Ala Glu Glu Ala Ala Gly Ser 
                165                 170                 175     


Leu Phe Cys Leu Ala Thr Gly Tyr Asn Ile Leu Asn Lys Leu Ser Lys 
            180                 185                 190         


Asn Lys Tyr Leu Lys Gln Asp Ser Asn Val Leu Ile Asn Gly Gly Thr 
        195                 200                 205             


Ser Ser Val Gly Met Phe Val Ile Gln Leu Leu Lys Arg His Tyr Lys 
    210                 215                 220                 


Leu Gln Lys Lys Leu Val Ile Val Thr Ser Ala Asn Gly Pro Gln Val 
225                 230                 235                 240 


Leu Gln Glu Lys Phe Pro Asp Leu Ala Asp Glu Met Ile Phe Ile Asp 
                245                 250                 255     


Tyr Leu Thr Cys Arg Gly Lys Ser Ser Lys Pro Leu Arg Lys Met Leu 
            260                 265                 270         


Glu Glu Lys Lys Ile Ser Gln Tyr Asp Pro Val Glu Asp Lys Glu Thr 
        275                 280                 285             


Ile Leu Asn Tyr Asn Glu Gly Lys Phe Asp Val Val Leu Asp Phe Val 
    290                 295                 300                 


Gly Gly Tyr Asp Ile Leu Ser His Ser Ser Ser Leu Ile His Gly Gly 
305                 310                 315                 320 


Gly Ala Tyr Val Thr Thr Val Gly Asp Tyr Val Ala Asn Tyr Lys Glu 
                325                 330                 335     


Asp Ile Phe Asp Ser Trp Asp Asn Pro Ser Ala Asn Ala Arg Lys Met 
            340                 345                 350         


Phe Gly Ser Ile Ile Trp Ser Tyr Asn Tyr Thr His Tyr Tyr Phe Asp 
        355                 360                 365             


Pro Asn Ala Lys Thr Ala Ser Ala Asn Asn Asp Trp Ile Glu Gln Cys 
    370                 375                 380                 


Gly Asp Phe Leu Lys Asn Gly Thr Val Lys Cys Val Val Asp Lys Val 
385                 390                 395                 400 


Tyr Asp Trp Lys Ile His Lys Glu Ala Phe Ser Tyr Met Ala Thr Gln 
                405                 410                 415     


Arg Ala Gln Gly Lys Leu Ile Met Asn Val Glu Lys Phe 
            420                 425                 


<210>  5
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  gRNA

<400>  5
ttattcctgg aaaaatttca                                                   20


<210>  6
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  gRNA

<400>  6
tataagaaaa tgcttcttta                                                   20


