               SEQUENCE LISTING

<110> DSM IP Assets B.V.

<120> VARIANT MALTOGENIC ALPHA-AMYLASE

<130> 32852-WO-PCT

<150> EP18165871.7
<151> 2018-04-05

<160> 5

<170> BiSSAP 1.3.6

<210> 1
<211> 686
<212> PRT
<213> Artificial Sequence


<220> 
<223> mature polypeptide of maltogenic alpha amylase derived from
      Bacillus stearothermophilus

<400> 1
Ser Ser Ser Ala Ser Val Lys Gly Asp Val Ile Tyr Gln Ile Ile Ile 
1               5                   10                  15      
Asp Arg Phe Tyr Asp Gly Asp Thr Thr Asn Asn Asn Pro Ala Lys Ser 
            20                  25                  30          
Tyr Gly Leu Tyr Asp Pro Thr Lys Ser Lys Trp Lys Met Tyr Trp Gly 
        35                  40                  45              
Gly Asp Leu Glu Gly Val Arg Gln Lys Leu Pro Tyr Leu Lys Gln Leu 
    50                  55                  60                  
Gly Val Thr Thr Ile Trp Leu Ser Pro Val Leu Asp Asn Leu Asp Thr 
65                  70                  75                  80  
Leu Ala Gly Thr Asp Asn Thr Gly Tyr His Gly Tyr Trp Thr Arg Asp 
                85                  90                  95      
Phe Lys Gln Ile Glu Glu His Phe Gly Asn Trp Thr Thr Phe Asp Thr 
            100                 105                 110         
Leu Val Asn Asp Ala His Gln Asn Gly Ile Lys Val Ile Val Asp Phe 
        115                 120                 125             
Val Pro Asn His Ser Thr Pro Phe Lys Ala Asn Asp Ser Thr Phe Ala 
    130                 135                 140                 
Glu Gly Gly Ala Leu Tyr Asn Asn Gly Thr Tyr Met Gly Asn Tyr Phe 
145                 150                 155                 160 
Asp Asp Ala Thr Lys Gly Tyr Phe His His Asn Gly Asp Ile Ser Asn 
                165                 170                 175     
Trp Asp Asp Arg Tyr Glu Ala Gln Trp Lys Asn Phe Thr Asp Pro Ala 
            180                 185                 190         
Gly Phe Ser Leu Ala Asp Leu Ser Gln Glu Asn Gly Thr Ile Ala Gln 
        195                 200                 205             
Tyr Leu Thr Asp Ala Ala Val Gln Leu Val Ala His Gly Ala Asp Gly 
    210                 215                 220                 
Leu Arg Ile Asp Ala Val Lys His Phe Asn Ser Gly Phe Ser Lys Ser 
225                 230                 235                 240 
Leu Ala Asp Lys Leu Tyr Gln Lys Lys Asp Ile Phe Leu Val Gly Glu 
                245                 250                 255     
Trp Tyr Gly Asp Asp Pro Gly Thr Ala Asn His Leu Glu Lys Val Arg 
            260                 265                 270         
Tyr Ala Asn Asn Ser Gly Val Asn Val Leu Asp Phe Asp Leu Asn Thr 
        275                 280                 285             
Val Ile Arg Asn Val Phe Gly Thr Phe Thr Gln Thr Met Tyr Asp Leu 
    290                 295                 300                 
Asn Asn Met Val Asn Gln Thr Gly Asn Glu Tyr Lys Tyr Lys Glu Asn 
305                 310                 315                 320 
Leu Ile Thr Phe Ile Asp Asn His Asp Met Ser Arg Phe Leu Ser Val 
                325                 330                 335     
Asn Ser Asn Lys Ala Asn Leu His Gln Ala Leu Ala Phe Ile Leu Thr 
            340                 345                 350         
Ser Arg Gly Thr Pro Ser Ile Tyr Tyr Gly Thr Glu Gln Tyr Met Ala 
        355                 360                 365             
Gly Gly Asn Asp Pro Tyr Asn Arg Gly Met Met Pro Ala Phe Asp Thr 
    370                 375                 380                 
Thr Thr Thr Ala Phe Lys Glu Val Ser Thr Leu Ala Gly Leu Arg Arg 
385                 390                 395                 400 
Asn Asn Ala Ala Ile Gln Tyr Gly Thr Thr Thr Gln Arg Trp Ile Asn 
                405                 410                 415     
Asn Asp Val Tyr Ile Tyr Glu Arg Lys Phe Phe Asn Asp Val Val Leu 
            420                 425                 430         
Val Ala Ile Asn Arg Asn Thr Gln Ser Ser Tyr Ser Ile Ser Gly Leu 
        435                 440                 445             
Gln Thr Ala Leu Pro Asn Gly Ser Tyr Ala Asp Tyr Leu Ser Gly Leu 
    450                 455                 460                 
Leu Gly Gly Asn Gly Ile Ser Val Ser Asn Gly Ser Val Ala Ser Phe 
465                 470                 475                 480 
Thr Leu Ala Pro Gly Ala Val Ser Val Trp Gln Tyr Ser Thr Ser Ala 
                485                 490                 495     
Ser Ala Pro Gln Ile Gly Ser Val Ala Pro Asn Met Gly Ile Pro Gly 
            500                 505                 510         
Asn Val Val Thr Ile Asp Gly Lys Gly Phe Gly Thr Thr Gln Gly Thr 
        515                 520                 525             
Val Thr Phe Gly Gly Val Thr Ala Thr Val Lys Ser Trp Thr Ser Asn 
    530                 535                 540                 
Arg Ile Glu Val Tyr Val Pro Asn Met Ala Ala Gly Leu Thr Asp Val 
545                 550                 555                 560 
Lys Val Thr Ala Gly Gly Val Ser Ser Asn Leu Tyr Ser Tyr Asn Ile 
                565                 570                 575     
Leu Ser Gly Thr Gln Thr Ser Val Val Phe Thr Val Lys Ser Ala Pro 
            580                 585                 590         
Pro Thr Asn Leu Gly Asp Lys Ile Tyr Leu Thr Gly Asn Ile Pro Glu 
        595                 600                 605             
Leu Gly Asn Trp Ser Thr Asp Thr Ser Gly Ala Val Asn Asn Ala Gln 
    610                 615                 620                 
Gly Pro Leu Leu Ala Pro Asn Tyr Pro Asp Trp Phe Tyr Val Phe Ser 
625                 630                 635                 640 
Val Pro Ala Gly Lys Thr Ile Gln Phe Lys Phe Phe Ile Lys Arg Ala 
                645                 650                 655     
Asp Gly Thr Ile Gln Trp Glu Asn Gly Ser Asn His Val Ala Thr Thr 
            660                 665                 670         
Pro Thr Gly Ala Thr Gly Asn Ile Thr Val Thr Trp Gln Asn 
        675                 680                 685     

<210> 2
<211> 2186
<212> DNA
<213> Artificial Sequence


<220> 
<223> ROM 1 module encoding a maltogenic alpha-amylase containing amino
      acid substitutions F188L, S200N, D261G and T288P

<400> 2
agcgcgtctc ctatgaaaaa gaaaacgctt tctttgttcg taggcctgat gctgctgatc      60

ggtttgctgt tctctggttc tcttccatac aacccaaacg ctgctgaagc aagctcaagt     120

gcttctgtaa aaggcgatgt gatttaccaa atcatcatcg accgtttcta tgacggagac     180

actactaaca acaacccagc aaaatcatac ggtctttacg atccgacaaa atcaaaatgg     240

aaaatgtact ggggcggaga tcttgaaggc gttcgtcaaa aacttcctta cctgaaacaa     300

ttaggtgtga cgacgatttg gctgtctcct gttcttgata accttgatac attggcaggt     360

actgacaaca ctggctacca cggctactgg acgagagact tcaaacaaat cgaagaacac     420

ttcggaaact ggacgacatt cgatacgctt gtgaacgatg ctcaccaaaa cggaatcaaa     480

gttatcgttg atttcgttcc aaaccacagc actccattca aagctaacga ttcaacgttc     540

gctgaaggcg gtgctcttta caacaacgga acttacatgg gcaactactt tgatgatgca     600

acaaaaggct acttccacca caacggagat atttctaact gggatgaccg ttatgaagct     660

caatggaaaa acctgactga tccggcaggc ttcagccttg ctgatcttaa ccaagaaaac     720

ggaacgattg ctcaatacct gacagacgct gctgttcaat tagttgctca cggtgcagac     780

ggacttcgca ttgacgctgt aaaacacttc aacagcggct tctcaaaatc acttgctgac     840

aaactttacc aaaagaaaga tattttcctt gttggtgaat ggtacggaga tggcccaggt     900

actgcaaacc accttgaaaa agttcgttac gcaaacaaca gcggagtaaa cgtgcttgat     960

ttcgatttaa acccagtgat ccgtaatgta ttcggaacat tcactcaaac gatgtacgat    1020

ttgaacaaca tggtgaacca aacaggaaat gaatacaaat acaaagaaaa cctgattaca    1080

ttcatcgaca accatgatat gtctcgtttc ctttctgtaa acagcaacaa agcaaacctt    1140

caccaagctc ttgcattcat tttaacttct cgcggaactc caagcattta ctacggaact    1200

gaacaataca tggcaggcgg aaatgatcct tacaaccgcg gaatgatgcc tgctttcgac    1260

acaacaacta ctgcattcaa agaagtatca acgcttgctg gtctaagacg caacaacgct    1320

gctattcaat acggaactac tactcaacgc tggatcaaca acgacgttta catctacgaa    1380

cgcaaattct tcaacgatgt tgtgcttgtt gcaatcaacc gtaatacaca atcttcttac    1440

agcatttctg gtcttcaaac ggctcttcct aacggttctt acgctgatta cctaagcgga    1500

cttcttggcg gaaacggcat ttctgtttca aacggttctg ttgcttcttt cacacttgct    1560

cctggtgctg tttctgtttg gcaatactct acttctgctt ctgctcctca aatcggttct    1620

gtagctccaa acatgggcat cccaggaaac gttgtgacga ttgacggaaa aggcttcgga    1680

acaactcaag gtactgtaac gttcggcggc gttactgcaa ctgtaaaaag ctggacttct    1740

aaccgtatcg aagtgtacgt gccgaacatg gctgctggtt tgactgatgt gaaagtaact    1800

gcaggcggcg tttcttcaaa cctttacagc tacaacatcc tttctggtac tcaaacttct    1860

gttgtgttta ctgtgaaatc agctccgccg acaaaccttg gtgataaaat ctacctgact    1920

ggaaacatcc cagagcttgg caactggagc actgatacaa gcggcgctgt taacaacgct    1980

caaggaccgc ttcttgctcc aaactaccct gactggttct acgtattctc tgttcctgct    2040

ggaaaaacaa tccaattcaa attctttatc aaacgtgctg acggcacgat tcaatgggaa    2100

aacggttcaa accatgtggc aacaactcca actggtgcaa ctggtaacat cactgttact    2160

tggcaaaact aataaggaga cgcgct                                         2186


<210> 3
<211> 283
<212> DNA
<213> Artificial Sequence


<220> 
<223> BsmBI site

<400> 3
agcgcgtctc cccgctaaaa attttacaaa aaggtattga ctttccctac agggtgtgta     60

ataatttaat tataaggaca aatgaataaa gattgtatcc ttcggggcag ggtggaaatc    120

ccgaccggcg gtagtaaagc acatttgctt tagagtccgt gacccgtgtg cataagcacg    180

cggtggattc agtttaagct gaagccgaca gtgaaagtct ggatgggaga aggatggacg    240

gtaaataaca aaagaaagga ggtgatcata tgggagacgc gct                      283


<210> 4
<211> 30
<212> PRT
<213> Artificial Sequence


<220> 
<223> forward primer

<400> 4
Cys Ala Ala Ala Cys Ala Thr Ala Thr Ala Gly Ala Gly Cys Thr Thr 
1               5                   10                  15      
Cys Cys Ala Gly Ala Thr Gly Ala Thr Gly Gly Cys Ala Gly 
            20                  25                  30  

<210> 5
<211> 27
<212> DNA
<213> Artificial Sequence


<220> 
<223> Reverse primer

<400> 5
ggtcatccca gttagaaata tctccgt                                        27


