               SEQUENCE LISTING

<110> DSM IP Assets B.V.

<120> LIPOLYTIC ENZYME VARIANTS

<130> 32949-WO-PCT

<150> EP18178445.5
<151> 2018-06-19

<160> 7

<170> BiSSAP 1.3.6

<210> 1
<211> 346
<212> PRT
<213> Artificial Sequence


<220> 
<223> Amino acid sequence of a polypeptide having lipase activity

<400> 1
Met Leu Leu Leu Ser Leu Leu Ser Ile Val Thr Leu Ala Val Ala Ser 
1               5                   10                  15      
Pro Leu Ser Val Glu Glu Tyr Ala Lys Ala Leu Glu Glu Arg Ala Val 
            20                  25                  30          
Thr Val Ser Ser Ser Glu Leu Asn Asn Phe Lys Phe Tyr Ile Gln His 
        35                  40                  45              
Gly Ala Ala Ala Tyr Cys Asn Ser Glu Thr Ala Ala Gly Ala Asn Val 
    50                  55                  60                  
Thr Cys Thr Gly Asn Ala Cys Pro Glu Ile Glu Ala Asn Gly Val Thr 
65                  70                  75                  80  
Val Val Ala Ser Phe Thr Gly Thr Lys Thr Gly Ile Gly Gly Tyr Val 
                85                  90                  95      
Ser Thr Asp Asn Thr Asn Lys Glu Ile Val Leu Ser Phe Arg Gly Ser 
            100                 105                 110         
Ile Asn Ile Arg Asn Trp Leu Thr Asn Leu Asp Phe Gly Gln Asp Asp 
        115                 120                 125             
Cys Ser Leu Thr Ser Gly Cys Gly Val His Ser Gly Phe Gln Arg Ala 
    130                 135                 140                 
Trp Glu Glu Ile Ala Asp Asn Leu Thr Ala Ala Val Ala Lys Ala Lys 
145                 150                 155                 160 
Thr Ala Asn Pro Asp Tyr Lys Val Val Ala Thr Gly His Ser Leu Gly 
                165                 170                 175     
Gly Ala Val Ala Thr Leu Ala Gly Ala Asn Leu Arg Ala Ala Gly Thr 
            180                 185                 190         
Pro Leu Asp Ile Tyr Thr Tyr Gly Ser Pro Arg Val Gly Asn Ala Glu 
        195                 200                 205             
Leu Ala Glu Phe Ile Ser Asn Gln Thr Gly Gly Glu Phe Arg Val Thr 
    210                 215                 220                 
His Gly Asp Asp Pro Val Pro Arg Leu Pro Pro Leu Ile Phe Gly Tyr 
225                 230                 235                 240 
Arg His Thr Ser Pro Glu Tyr Trp Leu Asp Gly Ser Gly Gly Asp Lys 
                245                 250                 255     
Ile Asn Tyr Thr Ile Asn Asp Ile Lys Val Cys Glu Gly Ala Ala Asn 
            260                 265                 270         
Leu Gln Cys Asn Gly Gly Thr Leu Gly Leu Asp Ile Ala Ala His Leu 
        275                 280                 285             
His Tyr Phe Gln Ala Thr Asp Ala Cys Asn Ala Gly Gly Phe Ser Trp 
    290                 295                 300                 
Arg Arg Tyr Arg Ser Ala Glu Ser Val Asp Lys Arg Ala Thr Met Thr 
305                 310                 315                 320 
Asp Ala Glu Leu Glu Lys Lys Leu Asn Ser Tyr Val Gln Met Asp Lys 
                325                 330                 335     
Glu Tyr Val Lys Asn Asn Gln Ala Arg Ser 
            340                 345     

<210> 2
<211> 1041
<212> DNA
<213> Artificial Sequence


<220> 
<223> Nucleotide sequence encoding a lipase of SEQ ID NO: 1

<400> 2
atgcttctcc tctccctcct ctccattgtc accctcgctg ttgcttctcc tctgtccgtt      60

gaggagtacg ccaaggccct cgaggagcgt gccgtcaccg tctcctcctc cgagctcaac     120

aacttcaagt tctacatcca gcacggtgct gctgcctact gcaactccga gactgctgct     180

ggtgccaacg tcacctgcac tggcaacgcc tgccccgaga ttgaggccaa cggtgtcacc     240

gttgttgcct ccttcactgg taccaagact ggtatcggtg gctacgtctc caccgacaac     300

accaacaagg agatcgtcct ttctttccgt ggcagcatca acatccgcaa ctggctgacc     360

aacctggact tcggccagga tgactgctct ctgacctccg gctgcggtgt ccactccggt     420

ttccagcgtg cctgggagga gattgccgac aacctgaccg ctgctgttgc caaggccaag     480

actgccaacc ccgactacaa ggttgttgcc actggccact ccctgggtgg tgctgttgcc     540

accctggctg gtgccaacct ccgtgctgct ggtacccccc tcgacatcta cacctacggc     600

tctccccgtg tcggcaacgc cgagcttgct gagttcatct ccaaccagac tggtggtgag     660

ttccgtgtca cccacggtga tgaccccgtc ccccgtcttc ctcctctgat cttcggctac     720

cgccacacct cccccgagta ctggctcgat ggcagcggtg gtgacaagat caactacacc     780

atcaacgaca tcaaggtctg cgagggtgct gccaacctgc agtgcaacgg tggtaccctg     840

ggactcgaca ttgctgctca cctgcactac ttccaggcca ctgatgcctg caacgccggt     900

ggtttcagct ggcgccgcta ccgctctgct gagagcgttg acaagcgtgc caccatgact     960

gatgctgagc tcgagaagaa gctcaacagc tacgtgcaga tggacaagga gtacgtcaag    1020

aacaaccagg ctcgctccta a                                              1041


<210> 3
<211> 348
<212> PRT
<213> Artificial Sequence


<220> 
<223> Amino acid sequence of a lipase of SEQ ID NO: 1 containing two
      proline residues at position 304

<400> 3
Met Leu Leu Leu Ser Leu Leu Ser Ile Val Thr Leu Ala Val Ala Ser 
1               5                   10                  15      
Pro Leu Ser Val Glu Glu Tyr Ala Lys Ala Leu Glu Glu Arg Ala Val 
            20                  25                  30          
Thr Val Ser Ser Ser Glu Leu Asn Asn Phe Lys Phe Tyr Ile Gln His 
        35                  40                  45              
Gly Ala Ala Ala Tyr Cys Asn Ser Glu Thr Ala Ala Gly Ala Asn Val 
    50                  55                  60                  
Thr Cys Thr Gly Asn Ala Cys Pro Glu Ile Glu Ala Asn Gly Val Thr 
65                  70                  75                  80  
Val Val Ala Ser Phe Thr Gly Thr Lys Thr Gly Ile Gly Gly Tyr Val 
                85                  90                  95      
Ser Thr Asp Asn Thr Asn Lys Glu Ile Val Leu Ser Phe Arg Gly Ser 
            100                 105                 110         
Ile Asn Ile Arg Asn Trp Leu Thr Asn Leu Asp Phe Gly Gln Asp Asp 
        115                 120                 125             
Cys Ser Leu Thr Ser Gly Cys Gly Val His Ser Gly Phe Gln Arg Ala 
    130                 135                 140                 
Trp Glu Glu Ile Ala Asp Asn Leu Thr Ala Ala Val Ala Lys Ala Lys 
145                 150                 155                 160 
Thr Ala Asn Pro Asp Tyr Lys Val Val Ala Thr Gly His Ser Leu Gly 
                165                 170                 175     
Gly Ala Val Ala Thr Leu Ala Gly Ala Asn Leu Arg Ala Ala Gly Thr 
            180                 185                 190         
Pro Leu Asp Ile Tyr Thr Tyr Gly Ser Pro Arg Val Gly Asn Ala Glu 
        195                 200                 205             
Leu Ala Glu Phe Ile Ser Asn Gln Thr Gly Gly Glu Phe Arg Val Thr 
    210                 215                 220                 
His Gly Asp Asp Pro Val Pro Arg Leu Pro Pro Leu Ile Phe Gly Tyr 
225                 230                 235                 240 
Arg His Thr Ser Pro Glu Tyr Trp Leu Asp Gly Ser Gly Gly Asp Lys 
                245                 250                 255     
Ile Asn Tyr Thr Ile Asn Asp Ile Lys Val Cys Glu Gly Ala Ala Asn 
            260                 265                 270         
Leu Gln Cys Asn Gly Gly Thr Leu Gly Leu Asp Ile Ala Ala His Leu 
        275                 280                 285             
His Tyr Phe Gln Ala Thr Asp Ala Cys Asn Ala Gly Gly Phe Ser Trp 
    290                 295                 300                 
Pro Pro Arg Arg Tyr Arg Ser Ala Glu Ser Val Asp Lys Arg Ala Thr 
305                 310                 315                 320 
Met Thr Asp Ala Glu Leu Glu Lys Lys Leu Asn Ser Tyr Val Gln Met 
                325                 330                 335     
Asp Lys Glu Tyr Val Lys Asn Asn Gln Ala Arg Ser 
            340                 345             

<210> 4
<211> 1047
<212> DNA
<213> Artificial Sequence


<220> 
<223> Nucleotide sequence encoding a lipase according to SEQ ID NO: 3

<400> 4
atgcttctcc tctccctcct ctccattgtc accctcgctg ttgcttctcc tctgtccgtt      60

gaggagtacg ccaaggccct cgaggagcgt gccgtcaccg tctcctcctc cgagctcaac     120

aacttcaagt tctacatcca gcacggtgct gctgcctact gcaactccga gactgctgct     180

ggtgccaacg tcacctgcac tggcaacgcc tgccccgaga ttgaggccaa cggtgtcacc     240

gttgttgcct ccttcactgg taccaagact ggtatcggtg gctacgtctc caccgacaac     300

accaacaagg agatcgtcct ttctttccgt ggcagcatca acatccgcaa ctggctgacc     360

aacctggact tcggccagga tgactgctct ctgacctccg gctgcggtgt ccactccggt     420

ttccagcgtg cctgggagga gattgccgac aacctgaccg ctgctgttgc caaggccaag     480

actgccaacc ccgactacaa ggttgttgcc actggccact ccctgggtgg tgctgttgcc     540

accctggctg gtgccaacct ccgtgctgct ggtacccccc tcgacatcta cacctacggc     600

tctccccgtg tcggcaacgc cgagcttgct gagttcatct ccaaccagac tggtggtgag     660

ttccgtgtca cccacggtga tgaccccgtc ccccgtcttc ctcctctgat cttcggctac     720

cgccacacct cccccgagta ctggctcgat ggcagcggtg gtgacaagat caactacacc     780

atcaacgaca tcaaggtctg cgagggtgct gccaacctgc agtgcaacgg tggtaccctg     840

ggactcgaca ttgctgctca cctgcactac ttccaggcca ctgatgcctg caacgccggt     900

ggtttcagct ggcctccccg ccgctaccgc tctgctgaga gcgttgacaa gcgtgccacc     960

atgactgatg ctgagctcga gaagaagctc aacagctacg tgcagatgga caaggagtac    1020

gtcaagaaca accaggctcg ctcctaa                                        1047


<210> 5
<211> 13
<212> DNA
<213> Artificial Sequence


<220> 
<223> Modified translational initiation sequence of glucoamylase glA
      promotor

<400> 5
caccgtcaaa atg                                                       13


<210> 6
<211> 346
<212> PRT
<213> Artificial Sequence


<220> 
<223> Fusarium oxysporum

<400> 6
Met Leu Leu Leu Pro Leu Leu Ser Ala Ile Thr Leu Ala Val Ala Ser 
1               5                   10                  15      
Pro Val Ala Leu Asp Asp Tyr Val Asn Ser Leu Glu Glu Arg Ala Val 
            20                  25                  30          
Gly Val Thr Thr Thr Asp Phe Ser Asn Phe Lys Phe Tyr Ile Gln His 
        35                  40                  45              
Gly Ala Ala Ala Tyr Cys Asn Ser Glu Ala Ala Ala Gly Ser Lys Ile 
    50                  55                  60                  
Thr Cys Ser Asn Asn Gly Cys Pro Thr Val Gln Gly Asn Gly Ala Thr 
65                  70                  75                  80  
Ile Val Thr Ser Phe Val Gly Ser Lys Thr Gly Ile Gly Gly Tyr Val 
                85                  90                  95      
Ala Thr Asp Ser Ala Arg Lys Glu Ile Val Val Ser Phe Arg Gly Ser 
            100                 105                 110         
Ile Asn Ile Arg Asn Trp Leu Thr Asn Leu Asp Phe Gly Gln Glu Asp 
        115                 120                 125             
Cys Ser Leu Val Ser Gly Cys Gly Val His Ser Gly Phe Gln Arg Ala 
    130                 135                 140                 
Trp Asn Glu Ile Ser Ser Gln Ala Thr Ala Ala Val Ala Ser Ala Arg 
145                 150                 155                 160 
Lys Ala Asn Pro Ser Phe Asn Val Ile Ser Thr Gly His Ser Leu Gly 
                165                 170                 175     
Gly Ala Val Ala Val Leu Ala Ala Ala Asn Leu Arg Val Gly Gly Thr 
            180                 185                 190         
Pro Val Asp Ile Tyr Thr Tyr Gly Ser Pro Arg Val Gly Asn Ala Gln 
        195                 200                 205             
Leu Ser Ala Phe Val Ser Asn Gln Ala Gly Gly Glu Tyr Arg Val Thr 
    210                 215                 220                 
His Ala Asp Asp Pro Val Pro Arg Leu Pro Pro Leu Ile Phe Gly Tyr 
225                 230                 235                 240 
Arg His Thr Thr Pro Glu Phe Trp Leu Ser Gly Gly Gly Gly Asp Lys 
                245                 250                 255     
Val Asp Tyr Thr Ile Ser Asp Val Lys Val Cys Glu Gly Ala Ala Asn 
            260                 265                 270         
Leu Gly Cys Asn Gly Gly Thr Leu Gly Leu Asp Ile Ala Ala His Leu 
        275                 280                 285             
His Tyr Phe Gln Ala Thr Asp Ala Cys Asn Ala Gly Gly Phe Ser Trp 
    290                 295                 300                 
Arg Arg Tyr Arg Ser Ala Glu Ser Val Asp Lys Arg Ala Thr Met Thr 
305                 310                 315                 320 
Asp Ala Glu Leu Glu Lys Lys Leu Asn Ser Tyr Val Gln Met Asp Lys 
                325                 330                 335     
Glu Tyr Val Lys Asn Asn Gln Ala Arg Ser 
            340                 345     

<210> 7
<211> 1041
<212> DNA
<213> Artificial Sequence


<220> 
<223> Nucleotide sequence encoding Fusarium oxysporum lipase that was
      codon-pair optimized for expression in Aspergillus niger

<400> 7
atgctcctcc ttcctctcct ctccgccatc accctggccg ttgcctcccc cgttgctctc      60

gatgactacg tcaactccct cgaggagcgt gccgtcggtg tcaccaccac cgacttctcc     120

aacttcaagt tctacatcca gcacggtgct gctgcctact gcaactccga ggctgctgct     180

ggcagcaaga tcacctgctc caacaacggc tgccccaccg tccagggcaa cggtgccacc     240

attgtcacct ccttcgtcgg cagcaagact ggtatcggtg gctacgttgc caccgactct     300

gctcgcaagg agattgtcgt ctccttccgt ggcagcatca acatccgcaa ctggctgacc     360

aacctggact tcggccagga agactgctct cttgtctccg gctgcggtgt ccactccggt     420

ttccagcgtg cctggaacga gatctcctcc caggccactg ctgccgttgc ttctgctcgc     480

aaggccaacc cctccttcaa cgtcatctcc actggccact ctcttggtgg tgctgttgct     540

gtccttgctg ctgccaacct ccgtgtcggt ggtactcccg ttgacatcta cacctacggc     600

agccctcgtg tcggcaacgc ccagctgtct gctttcgtct ccaaccaggc tggtggtgaa     660

taccgtgtca cccacgccga tgaccccgtt ccccgtcttc ctcctctgat cttcggttac     720

cgccacacca ctcccgagtt ctggctatcc ggtggtggtg gtgacaaggt cgactacacc     780

atctccgatg tcaaggtctg cgagggtgct gccaacctgg gctgcaacgg tggtaccctg     840

ggactcgaca ttgctgctca cctgcactac ttccaggcca ctgatgcctg caacgccggt     900

ggtttcagct ggcgccgcta ccgctctgct gagagcgttg acaagcgtgc caccatgact     960

gatgctgagc tcgagaagaa gctcaacagc tacgtgcaga tggacaagga gtacgtcaag    1020

aacaaccagg ctcgctccta a                                              1041


