               SEQUENCE LISTING

<110> DSM IP Assets B.V.

<120> Proline specific endoprotease

<130> 30254-WO-PCT

<160> 6

<170> BiSSAP 1.2

<210> 1
<211> 1580
<212> DNA
<213> Artificial Sequence

<220> 
<221> source
<222> 1..1580
<223> /organism="Artificial Sequence"
      /note="Codon optimized nucleic acid encoding proline-specific
      endoprotease from Rasamsonia emersonii BC2G079 for expression in
      A. niger"
      /mol_type="unassigned DNA"

<400> 1
atgccctccc tctcctccct cgttgccttg actgcttctc ttgtctctct ggctgctgcc      60

gctgctcctc gtctccctct tcctcctcgc cctcccttgc ctccccgtga ccccttgcac     120

ggacctacca acgcctccgc cactttccag cagctcatcg accacaacca ccccgagctt     180

ggcaccttct cccagcgcta ctggtggaat gatgagttct ggaagggtcc cggctctccc     240

gttgtccttt tcacccccgg tgaagaagat gccagcggtt acgtgggcta cctgaagaac     300

accaccatca ccggtctgat cgctcagacc atcggtggtg ccgtcatcgt cctcgaacac     360

cgctactggg gccagtcctc cccctacgac tctctgacca ccaagaacct gcagtacctg     420

accctcaagc agtccattgc cgacctcacc tacttcgcca agaccgtcaa gctccccttc     480

gaccgcaacg gcagctccaa cgccgacaag gctccctggg ttctcagcgg tggaagctac     540

tctggtgctc tctccgcctg gactgccagc acctcccccg gtactttctg ggcctaccac     600

gccagctctg ctcctgttga ggccatctac gattactggc agtacttcgc tcccgtgcag     660

gatggattgc ctgccaactg ctcgaaggac ctctcccgtg tcgtcgacta catcgactcc     720

gttctccagt ccggcaacgc cactgccaag caacagctca aggacctttt cggtctgggt     780

gctctggagc acgacgatga cttcgcctcc gctcttgaga acggcccttg gctctggcag     840

tcgaactcgt tctacgaccc ctaccctcct gtctacgagt tctgcgacta cgttgagaac     900

gcctacgcca gccctcccgt tgctgctggt cccgatggtg ttggtctgga gaaggctctg     960

tctggctacg ccacctggtg gaacaaggtc ttcttccccg gctactgcgc tacctacggc    1020

tactggtcct ccaacgactc cattgcctgc ttcgacacct acaaccagtc gtcgcccatg    1080

ttcaccgacc tttccgtctc caacactatc aaccgccagt ggaactggtt cctctgcaac    1140

gagcccttct tctactggca ggatggtgct cccaagaacg tccccagcat tgtctctcgt    1200

ctggtcactg ctgagtactg gcagcgccag tgccccttgt tcttccctga agaggatggc    1260

tacacctacg gaagcgccaa gggcaagact gctgccgatg tcaacgcctg gaccaagggc    1320

tggttcttga ctaacaccac ccgtctgatc tggaccaacg gcgagcttga cccctggcgc    1380

tctgctggtg tcagcagcaa gttccgtccc ggtggtcccc tccagtccac tccccaggct    1440

cctctgcagc tcattcccga gggtgtccac tgctacgatc tgatcctcaa gaacgccgag    1500

gccaacgccg gtgtgcagcg tgttgtcacc aacgaggttg ctcagatcaa ggcctgggtg    1560

aacgaatact accgtaagta                                                1580


<210> 2
<211> 526
<212> PRT
<213> Artificial Sequence

<220> 
<223> Rasamsonia emersonii

<400> 2
Met Pro Ser Leu Ser Ser Leu Val Ala Leu Thr Ala Ser Leu Val Ser 
1               5                   10                  15      
Leu Ala Ala Ala Ala Ala Pro Arg Leu Pro Leu Pro Pro Arg Pro Pro 
            20                  25                  30          
Leu Pro Pro Arg Asp Pro Leu His Gly Pro Thr Asn Ala Ser Ala Thr 
        35                  40                  45              
Phe Gln Gln Leu Ile Asp His Asn His Pro Glu Leu Gly Thr Phe Ser 
    50                  55                  60                  
Gln Arg Tyr Trp Trp Asn Asp Glu Phe Trp Lys Gly Pro Gly Ser Pro 
65                  70                  75                  80  
Val Val Leu Phe Thr Pro Gly Glu Glu Asp Ala Ser Gly Tyr Val Gly 
                85                  90                  95      
Tyr Leu Lys Asn Thr Thr Ile Thr Gly Leu Ile Ala Gln Thr Ile Gly 
            100                 105                 110         
Gly Ala Val Ile Val Leu Glu His Arg Tyr Trp Gly Gln Ser Ser Pro 
        115                 120                 125             
Tyr Asp Ser Leu Thr Thr Lys Asn Leu Gln Tyr Leu Thr Leu Lys Gln 
    130                 135                 140                 
Ser Ile Ala Asp Leu Thr Tyr Phe Ala Lys Thr Val Lys Leu Pro Phe 
145                 150                 155                 160 
Asp Arg Asn Gly Ser Ser Asn Ala Asp Lys Ala Pro Trp Val Leu Ser 
                165                 170                 175     
Gly Gly Ser Tyr Ser Gly Ala Leu Ser Ala Trp Thr Ala Ser Thr Ser 
            180                 185                 190         
Pro Gly Thr Phe Trp Ala Tyr His Ala Ser Ser Ala Pro Val Glu Ala 
        195                 200                 205             
Ile Tyr Asp Tyr Trp Gln Tyr Phe Ala Pro Val Gln Asp Gly Leu Pro 
    210                 215                 220                 
Ala Asn Cys Ser Lys Asp Leu Ser Arg Val Val Asp Tyr Ile Asp Ser 
225                 230                 235                 240 
Val Leu Gln Ser Gly Asn Ala Thr Ala Lys Gln Gln Leu Lys Asp Leu 
                245                 250                 255     
Phe Gly Leu Gly Ala Leu Glu His Asp Asp Asp Phe Ala Ser Ala Leu 
            260                 265                 270         
Glu Asn Gly Pro Trp Leu Trp Gln Ser Asn Ser Phe Tyr Asp Pro Tyr 
        275                 280                 285             
Pro Pro Val Tyr Glu Phe Cys Asp Tyr Val Glu Asn Ala Tyr Ala Ser 
    290                 295                 300                 
Pro Pro Val Ala Ala Gly Pro Asp Gly Val Gly Leu Glu Lys Ala Leu 
305                 310                 315                 320 
Ser Gly Tyr Ala Thr Trp Trp Asn Lys Val Phe Phe Pro Gly Tyr Cys 
                325                 330                 335     
Ala Thr Tyr Gly Tyr Trp Ser Ser Asn Asp Ser Ile Ala Cys Phe Asp 
            340                 345                 350         
Thr Tyr Asn Gln Ser Ser Pro Met Phe Thr Asp Leu Ser Val Ser Asn 
        355                 360                 365             
Thr Ile Asn Arg Gln Trp Asn Trp Phe Leu Cys Asn Glu Pro Phe Phe 
    370                 375                 380                 
Tyr Trp Gln Asp Gly Ala Pro Lys Asn Val Pro Ser Ile Val Ser Arg 
385                 390                 395                 400 
Leu Val Thr Ala Glu Tyr Trp Gln Arg Gln Cys Pro Leu Phe Phe Pro 
                405                 410                 415     
Glu Glu Asp Gly Tyr Thr Tyr Gly Ser Ala Lys Gly Lys Thr Ala Ala 
            420                 425                 430         
Asp Val Asn Ala Trp Thr Lys Gly Trp Phe Leu Thr Asn Thr Thr Arg 
        435                 440                 445             
Leu Ile Trp Thr Asn Gly Glu Leu Asp Pro Trp Arg Ser Ala Gly Val 
    450                 455                 460                 
Ser Ser Lys Phe Arg Pro Gly Gly Pro Leu Gln Ser Thr Pro Gln Ala 
465                 470                 475                 480 
Pro Leu Gln Leu Ile Pro Glu Gly Val His Cys Tyr Asp Leu Ile Leu 
                485                 490                 495     
Lys Asn Ala Glu Ala Asn Ala Gly Val Gln Arg Val Val Thr Asn Glu 
            500                 505                 510         
Val Ala Gln Ile Lys Ala Trp Val Asn Glu Tyr Tyr Arg Lys 
        515                 520                 525     

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

<220> 
<223> Signal sequence of A. niger pectinemethylesterase 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 A.niger 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      
Lys 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


