               SEQUENCE LISTING

<110> DSM IP Assets B.V.

<120> Proline specific endoprotease

<130> 30252-WO-PCT

<160> 6

<170> BiSSAP 1.2

<210> 1
<211> 1565
<212> DNA
<213> Artificial Sequence

<220> 
<221> source
<222> 1..1565
<223> /organism="Artificial Sequence"
      /note="Codon optimized nucleic acid encoding proline-specific
      endoprotease BC2G075_Aspergillus_carbonarius, for expression in
      A. niger"
      /mol_type="unassigned DNA"

<400> 1
atggtcaagt ccatcctggc ctccgtcttc ttcgctgcca ctgctcttgc tgctcgtcct      60

cgcttagtgc ccaagcccgt gtctcgcccc gcctccagca cttctgccgc caccaccggt     120

gaggcctact tcgagcagct ggttgaccac cacaaccccg agaagggcac cttctcccag     180

cgctactggt ggagcactga gtactggggt ggtcccggct cccccgtcgt cctcttcacc     240

cccggtgaag tctctgccga tggctacgag ggctacctga ccaacgacac cctgaccggt     300

gtctacgctc aggagatcca gggtgctgtt gtgttgattg agcaccgtta ctggggcgac     360

agcagcccct acgaggtcct caacgccgag actctccagt acctgaccct cgaccaggct     420

gtccttgaca tgacctactt cgctgagact gtcaagttcc agttcgacaa ctcgacccgc     480

agcaacgccc agaacgctcc ttgggtgatg gtcggtggaa gctactctgg tgctctcacc     540

gcctgggttg agtccgtcgc tcctggaacc ttctgggcct accacgccac ctcggctcct     600

gttgaggcca tctacgactt ctggcagtac ttctacccca tctcccaggg tatggcccag     660

aactgctcca aggatgtctc ccgtgttgct gagcacgttg acaaggtcgg caagtctggc     720

actgctgagg agcagcagaa gctcaaggaa ctcttcggtc ttggtgctct tgagcactac     780

gatgacttcg ctgctgtcct ccccaacggc ccctacctct ggcaggacaa cgacttcgcc     840

actggatact ccgagttctt ccagttctgc gatgccgtcg agggtgtgga agccggtgct     900

gctgtgaccc ccggccccga gggtgttggt cttgagaagg ctctggccaa ctacgcctac     960

tggttcaact cgactctcct tcccaactac tgcgcttcct acggctactg gtcggatgag    1020

tggtccgttg cctgcttcga ctcctacaac gcctcctctc ctctgttcac cgacacctcc    1080

gttgacaacg ccgttgaccg tcagtgggaa tggttcctct gcaacgaacc tttcttctgg    1140

tggcaggatg gtgctcccga ggatgtcacc accattgtgc ctcgtctggt caacgcggaa    1200

tactggcagc gtcagtgctc tctgtacttc cccgaaacca acggctacac cttcggttcc    1260

gccaagaaca agactgctgc caccgtcaac gactggactg gtggctggtt cgaaacccgc    1320

aacaccaccc gtctcatctg gaccaacggc cagtacgacc cctggcgcga cagcggtgtc    1380

tcttccacct tccgtcctgg tggccagctc gtcagcactg ccaacgagcc tgtccagatc    1440

atccccggtg gtttccactg ctcggatctg tacatggccg actactacgc caacgccggt    1500

gtccgcaagg tcgtcgacaa cgaggttgct cagatcaaga agtgggttgc tgagtactac    1560

gccta                                                                1565


<210> 2
<211> 521
<212> PRT
<213> Artificial Sequence

<220> 
<223> Proline-specific endoprotease BC2G075_Aspergillus_carbonarius
      with PMeA signal sequence from A. niger

<400> 2
Met Val Lys Ser Ile Leu Ala Ser Val Phe Phe Ala Ala Thr Ala Leu 
1               5                   10                  15      
Ala Ala Arg Pro Arg Leu Val Pro Lys Pro Val Ser Arg Pro Ala Ser 
            20                  25                  30          
Ser Thr Ser Ala Ala Thr Thr Gly Glu Ala Tyr Phe Glu Gln Leu Val 
        35                  40                  45              
Asp His His Asn Pro Glu Lys Gly Thr Phe Ser Gln Arg Tyr Trp Trp 
    50                  55                  60                  
Ser Thr Glu Tyr Trp Gly Gly Pro Gly Ser Pro Val Val Leu Phe Thr 
65                  70                  75                  80  
Pro Gly Glu Val Ser Ala Asp Gly Tyr Glu Gly Tyr Leu Thr Asn Asp 
                85                  90                  95      
Thr Leu Thr Gly Val Tyr Ala Gln Glu Ile Gln Gly Ala Val Val Leu 
            100                 105                 110         
Ile Glu His Arg Tyr Trp Gly Asp Ser Ser Pro Tyr Glu Val Leu Asn 
        115                 120                 125             
Ala Glu Thr Leu Gln Tyr Leu Thr Leu Asp Gln Ala Val Leu Asp Met 
    130                 135                 140                 
Thr Tyr Phe Ala Glu Thr Val Lys Phe Gln Phe Asp Asn Ser Thr Arg 
145                 150                 155                 160 
Ser Asn Ala Gln Asn Ala Pro Trp Val Met Val Gly Gly Ser Tyr Ser 
                165                 170                 175     
Gly Ala Leu Thr Ala Trp Val Glu Ser Val Ala Pro Gly Thr Phe Trp 
            180                 185                 190         
Ala Tyr His Ala Thr Ser Ala Pro Val Glu Ala Ile Tyr Asp Phe Trp 
        195                 200                 205             
Gln Tyr Phe Tyr Pro Ile Ser Gln Gly Met Ala Gln Asn Cys Ser Lys 
    210                 215                 220                 
Asp Val Ser Arg Val Ala Glu His Val Asp Lys Val Gly Lys Ser Gly 
225                 230                 235                 240 
Thr Ala Glu Glu Gln Gln Lys Leu Lys Glu Leu Phe Gly Leu Gly Ala 
                245                 250                 255     
Leu Glu His Tyr Asp Asp Phe Ala Ala Val Leu Pro Asn Gly Pro Tyr 
            260                 265                 270         
Leu Trp Gln Asp Asn Asp Phe Ala Thr Gly Tyr Ser Glu Phe Phe Gln 
        275                 280                 285             
Phe Cys Asp Ala Val Glu Gly Val Glu Ala Gly Ala Ala Val Thr Pro 
    290                 295                 300                 
Gly Pro Glu Gly Val Gly Leu Glu Lys Ala Leu Ala Asn Tyr Ala Tyr 
305                 310                 315                 320 
Trp Phe Asn Ser Thr Leu Leu Pro Asn Tyr Cys Ala Ser Tyr Gly Tyr 
                325                 330                 335     
Trp Ser Asp Glu Trp Ser Val Ala Cys Phe Asp Ser Tyr Asn Ala Ser 
            340                 345                 350         
Ser Pro Leu Phe Thr Asp Thr Ser Val Asp Asn Ala Val Asp Arg Gln 
        355                 360                 365             
Trp Glu Trp Phe Leu Cys Asn Glu Pro Phe Phe Trp Trp Gln Asp Gly 
    370                 375                 380                 
Ala Pro Glu Asp Val Thr Thr Ile Val Pro Arg Leu Val Asn Ala Glu 
385                 390                 395                 400 
Tyr Trp Gln Arg Gln Cys Ser Leu Tyr Phe Pro Glu Thr Asn Gly Tyr 
                405                 410                 415     
Thr Phe Gly Ser Ala Lys Asn Lys Thr Ala Ala Thr Val Asn Asp Trp 
            420                 425                 430         
Thr Gly Gly Trp Phe Glu Thr Arg Asn Thr Thr Arg Leu Ile Trp Thr 
        435                 440                 445             
Asn Gly Gln Tyr Asp Pro Trp Arg Asp Ser Gly Val Ser Ser Thr Phe 
    450                 455                 460                 
Arg Pro Gly Gly Gln Leu Val Ser Thr Ala Asn Glu Pro Val Gln Ile 
465                 470                 475                 480 
Ile Pro Gly Gly Phe His Cys Ser Asp Leu Tyr Met Ala Asp Tyr Tyr 
                485                 490                 495     
Ala Asn Ala Gly Val Arg Lys Val Val Asp Asn Glu Val Ala Gln Ile 
            500                 505                 510         
Lys Lys Trp Val Ala Glu Tyr Tyr Ala 
        515                 520     

<210> 3
<211> 17
<212> PRT
<213> Artificial Sequence

<220> 
<223> Signal sequence of A. niger pectineesterase PMeA ss

<400> 3
Met Val Lys Ser Ile Leu Ala Ser Val Phe Phe Ala Ala Thr Ala Leu 
1               5                   10                  15      
Ala 
    

<210> 4
<211> 19
<212> PRT
<213> Artificial Sequence

<220> 
<223> Pro-sequence of A.carbonarius proline-specific endoprotease

<400> 4
Ala Arg Pro Arg Leu Val Pro Lys Pro Val Ser Arg Pro Ala Ser Ser 
1               5                   10                  15      
Thr Ser Ala 
            

<210> 5
<211> 521
<212> PRT
<213> Artificial Sequence

<220> 
<223> Proline-specific endoprotease from Aspergillus niger comprising a
      PMeA signal sequence from A. niger

<400> 5
Met Val Lys Ser Ile Leu Ala Ser Val Phe Phe Ala Ala Thr Ala Leu 
1               5                   10                  15      
Ala Ala Arg Pro Arg Leu Val Pro Lys Pro Val Ser Arg Pro Ala Ser 
            20                  25                  30          
Ser Lys Ser Ala Ala Thr Thr Gly Glu Ala Tyr Phe Glu Gln Leu Leu 
        35                  40                  45              
Asp His His Asn Pro Glu Lys Gly Thr Phe Ser Gln Arg Tyr Trp Trp 
    50                  55                  60                  
Ser Thr Glu Tyr Trp Gly Gly Pro Gly Ser Pro Val Val Leu Phe Thr 
65                  70                  75                  80  
Pro Gly Glu Val Ser Ala Asp Gly Tyr Glu Gly Tyr Leu Thr Asn Glu 
                85                  90                  95      
Thr Leu Thr Gly Val Tyr Ala Gln Glu Ile Gln Gly Ala Val Ile Leu 
            100                 105                 110         
Ile Glu His Arg Tyr Trp Gly Asp Ser Ser Pro Tyr Glu Val Leu Asn 
        115                 120                 125             
Ala Glu Thr Leu Gln Tyr Leu Thr Leu Asp Gln Ala Ile Leu Asp Met 
    130                 135                 140                 
Thr Tyr Phe Ala Glu Thr Val Lys Leu Gln Phe Asp Asn Ser Thr Arg 
145                 150                 155                 160 
Ser Asn Ala Gln Asn Ala Pro Trp Val Met Val Gly Gly Ser Tyr Ser 
                165                 170                 175     
Gly Ala Leu Thr Ala Trp Thr Glu Ser Val Ala Pro Gly Thr Phe Trp 
            180                 185                 190         
Ala Tyr His Ala Thr Ser Ala Pro Val Glu Ala Ile Tyr Asp Tyr Trp 
        195                 200                 205             
Gln Tyr Phe Tyr Pro Ile Gln Gln Gly Met Ala Gln Asn Cys Ser Lys 
    210                 215                 220                 
Asp Val Ser Leu Val Ala Glu Tyr Val Asp Lys Ile Gly Lys Asn Gly 
225                 230                 235                 240 
Thr Ala Lys Glu Gln Gln Ala Leu Lys Glu Leu Phe Gly Leu Gly Ala 
                245                 250                 255     
Val Glu His Phe Asp Asp Phe Ala Ala Val Leu Pro Asn Gly Pro Tyr 
            260                 265                 270         
Leu Trp Gln Asp Asn Asp Phe Ala Thr Gly Tyr Ser Ser Phe Phe Gln 
        275                 280                 285             
Phe Cys Asp Ala Val Glu Gly Val Glu Ala Gly Ala Ala Val Thr Pro 
    290                 295                 300                 
Gly Pro Glu Gly Val Gly Leu Glu Lys Ala Leu Ala Asn Tyr Ala Asn 
305                 310                 315                 320 
Trp Phe Asn Ser Thr Ile Leu Pro Asp Tyr Cys Ala Ser Tyr Gly Tyr 
                325                 330                 335     
Trp Thr Asp Glu Trp Ser Val Ala Cys Phe Asp Ser Tyr Asn Ala Ser 
            340                 345                 350         
Ser Pro Ile Tyr Thr Asp Thr Ser Val Gly Asn Ala Val Asp Arg Gln 
        355                 360                 365             
Trp Glu Trp Phe Leu Cys Asn Glu Pro Phe Phe Tyr Trp Gln Asp Gly 
    370                 375                 380                 
Ala Pro Glu Gly Thr Ser Thr Ile Val Pro Arg Leu Val Ser Ala Ser 
385                 390                 395                 400 
Tyr Trp Gln Arg Gln Cys Pro Leu Tyr Phe Pro Glu Thr Asn Gly Tyr 
                405                 410                 415     
Thr Tyr Gly Ser Ala Lys Gly Lys Asn Ala Ala Thr Val Asn Ser Trp 
            420                 425                 430         
Thr Gly Gly Trp Asp Met Thr Arg Asn Thr Thr Arg Leu Ile Trp Thr 
        435                 440                 445             
Asn Gly Gln Tyr Asp Pro Trp Arg Asp Ser Gly Val Ser Ser Thr Phe 
    450                 455                 460                 
Arg Pro Gly Gly Pro Leu Ala Ser Thr Ala Asn Glu Pro Val Gln Ile 
465                 470                 475                 480 
Ile Pro Gly Gly Phe His Cys Ser Asp Leu Tyr Met Ala Asp Tyr Tyr 
                485                 490                 495     
Ala Asn Glu Gly Val Lys Lys Val Val Asp Asn Glu Val Lys Gln Ile 
            500                 505                 510         
Lys Glu Trp Val Glu Glu Tyr Tyr Ala 
        515                 520     

<210> 6
<211> 1566
<212> DNA
<213> Artificial Sequence

<220> 
<221> source
<222> 1..1566
<223> /organism="Artificial Sequence"
      /note="Proline-specific endoprotease from Aspergillus niger
      comprising a PMeA signal sequence from A. niger"
      /mol_type="unassigned DNA"

<400> 6
atggtcaagt ccatcctggc ctccgtcttc ttcgctgcca ctgctcttgc tgcaaggcct      60

cgtctcgttc ccaagcccgt ttctcgtccc gccagctcca agtccgctgc tactactggt     120

gaggcctact ttgaacagct gttggaccac cacaaccctg agaagggtac tttctcgcaa     180

agatactggt ggagcaccga gtactggggt ggtcccggat cccccgttgt cctgttcact     240

cccggtgagg tcagcgctga tggctacgag ggttatctga ccaacgagac tctcaccggt     300

gtctacgccc aggagattca gggtgctgtc atcctgatcg aacaccgata ctggggtgac     360

tcgtctccct acgaggtgct gaacgccgag actctccagt acttgaccct cgaccaggct     420

atccttgata tgacctactt cgccgaaacc gtcaagctcc agtttgacaa ctccacccgc     480

tccaacgctc agaacgctcc ttgggttatg gtcggcggca gctacagcgg tgctctgact     540

gcttggaccg agtccgttgc tcccggcacc ttctgggctt accacgccac ctctgctcct     600

gttgaggcca tctacgacta ctggcaatac ttctacccca ttcagcaggg tatggctcag     660

aactgctcca aagatgtctc tcttgtagca gaatacgtcg acaagatcgg caagaacggc     720

actgccaagg agcaacaggc tctgaaggag cttttcggcc taggagcagt ggagcacttc     780

gacgacttcg ccgctgttct gcccaacggt ccttacctct ggcaagacaa cgactttgcc     840

accggttact cttctttctt ccagttctgt gatgccgtcg agggtgtcga ggctggtgct     900

gccgtcaccc ccggtcctga aggtgttggt ctggaaaagg cccttgctaa ctacgcgaac     960

tggttcaact ctaccatcct ccccgattac tgcgccagct acggctactg gactgacgag    1020

tggtccgtcg cctgcttcga ctcctacaac gcctcctctc ctatatacac cgacaccagc    1080

gttggtaacg ccgtcgaccg tcagtgggag tggttcctct gcaatgagcc cttcttttac    1140

tggcaggacg gtgcccccga gggtacttca acgatagtac cccgcttagt gtccgcctcc    1200

tactggcagc gtcaatgtcc gttgtacttc cccgagacta acggttacac ctacggctcc    1260

gccaagggaa agaacgccgc caccgtcaac agctggaccg gtggctggga catgacccgt    1320

aacaccaccc gtctgatctg gacgaacggc caatacgacc cctggcgtga ctccggtgtc    1380

tcttccacct tccgccccgg tggtcccctc gcttcgaccg ccaacgagcc cgtccagata    1440

atacccggtg gtttccattg ctccgacctc tacatggcag actactacgc caacgagggc    1500

gtcaagaagg ttgtcgacaa cgaagtcaaa caaatcaagg agtgggttga ggaatactac    1560

gcgtaa                                                               1566


