                         SEQUENCE LISTING

<110>  DuPont Nutrition Biosciences ApS
       Cramer, Jacob Flyvholm
       Jensen, Lene Bojsen
       Kellett-Smith, Anja Hemmingsen
       Bladt, Tove
       Lee, Sang-Kye
 
<120>  Acetolactate Decarboxylase

<130>  NB40736 PCT

<150>  US62/165671
<151>  2015-05-22

<150>  US62/168406
<151>  2015-05-29

<150>  US62/166610
<151>  2015-05-26

<160>  8     

<170>  PatentIn version 3.5

<210>  1
<211>  858
<212>  DNA
<213>  Brevibacillus brevis

<400>  1
atgaaaaaaa atatcatcac ttctatcaca tctctggctc tggttgccgg gctgtctttg       60

actgcttttg cagctacaac ggctactgta ccagcaccac ctgccaagca ggaatccaaa      120

cctgcggttg ccgctaatcc ggcaccaaaa aatgtactgt ttcaatactc aacgatcaat      180

gcactcatgc ttggacagtt tgaaggggac ttgactttga aagacctgaa gctgcgaggc      240

gatatggggc ttggtaccat caatgatctc gatggagaga tgattcagat gggtacaaaa      300

ttctaccaga tcgacagcac cggaaaatta tcggagctgc cagaaagtgt gaaaactcca      360

tttgcggtta ctacacattt cgagccgaaa gaaaaaacta cattaaccaa tgtgcaagat      420

tacaatcaat taacaaaaat gcttgaggag aaatttgaaa acaagaacgt cttttatgcc      480

gtaaagctga ccggtacctt taagatggta aaggctagaa cagttccaaa acaaaccaga      540

ccttatccgc agctgactga agtaaccaaa aaacaatccg agtttgaatt taaaaatgtt      600

aagggaaccc tgattggctt ctatacgcca aattatgcag cagccctgaa tgttcccgga      660

ttccatctcc acttcatcac agaggataaa acaagtggcg gacacgtatt aaatctgcaa      720

tttgacaacg cgaatctgga aatttctccg atccatgagt ttgatgtaca attgccgcac      780

acagatgatt ttgcccactc tgatctgaca caagttacta ctagccaagt acaccaagct      840

gagtcagaaa gaaaataa                                                    858


<210>  2
<211>  285
<212>  PRT
<213>  Brevibacillus brevis

<400>  2

Met Lys Lys Asn Ile Ile Thr Ser Ile Thr Ser Leu Ala Leu Val Ala 
1               5                   10                  15      


Gly Leu Ser Leu Thr Ala Phe Ala Ala Thr Thr Ala Thr Val Pro Ala 
            20                  25                  30          


Pro Pro Ala Lys Gln Glu Ser Lys Pro Ala Val Ala Ala Asn Pro Ala 
        35                  40                  45              


Pro Lys Asn Val Leu Phe Gln Tyr Ser Thr Ile Asn Ala Leu Met Leu 
    50                  55                  60                  


Gly Gln Phe Glu Gly Asp Leu Thr Leu Lys Asp Leu Lys Leu Arg Gly 
65                  70                  75                  80  


Asp Met Gly Leu Gly Thr Ile Asn Asp Leu Asp Gly Glu Met Ile Gln 
                85                  90                  95      


Met Gly Thr Lys Phe Tyr Gln Ile Asp Ser Thr Gly Lys Leu Ser Glu 
            100                 105                 110         


Leu Pro Glu Ser Val Lys Thr Pro Phe Ala Val Thr Thr His Phe Glu 
        115                 120                 125             


Pro Lys Glu Lys Thr Thr Leu Thr Asn Val Gln Asp Tyr Asn Gln Leu 
    130                 135                 140                 


Thr Lys Met Leu Glu Glu Lys Phe Glu Asn Lys Asn Val Phe Tyr Ala 
145                 150                 155                 160 


Val Lys Leu Thr Gly Thr Phe Lys Met Val Lys Ala Arg Thr Val Pro 
                165                 170                 175     


Lys Gln Thr Arg Pro Tyr Pro Gln Leu Thr Glu Val Thr Lys Lys Gln 
            180                 185                 190         


Ser Glu Phe Glu Phe Lys Asn Val Lys Gly Thr Leu Ile Gly Phe Tyr 
        195                 200                 205             


Thr Pro Asn Tyr Ala Ala Ala Leu Asn Val Pro Gly Phe His Leu His 
    210                 215                 220                 


Phe Ile Thr Glu Asp Lys Thr Ser Gly Gly His Val Leu Asn Leu Gln 
225                 230                 235                 240 


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


Gln Leu Pro His Thr Asp Asp Phe Ala His Ser Asp Leu Thr Gln Val 
            260                 265                 270         


Thr Thr Ser Gln Val His Gln Ala Glu Ser Glu Arg Lys 
        275                 280                 285 


<210>  3
<211>  261
<212>  PRT
<213>  Brevibacillus brevis

<400>  3

Ala Thr Thr Ala Thr Val Pro Ala Pro Pro Ala Lys Gln Glu Ser Lys 
1               5                   10                  15      


Pro Ala Val Ala Ala Asn Pro Ala Pro Lys Asn Val Leu Phe Gln Tyr 
            20                  25                  30          


Ser Thr Ile Asn Ala Leu Met Leu Gly Gln Phe Glu Gly Asp Leu Thr 
        35                  40                  45              


Leu Lys Asp Leu Lys Leu Arg Gly Asp Met Gly Leu Gly Thr Ile Asn 
    50                  55                  60                  


Asp Leu Asp Gly Glu Met Ile Gln Met Gly Thr Lys Phe Tyr Gln Ile 
65                  70                  75                  80  


Asp Ser Thr Gly Lys Leu Ser Glu Leu Pro Glu Ser Val Lys Thr Pro 
                85                  90                  95      


Phe Ala Val Thr Thr His Phe Glu Pro Lys Glu Lys Thr Thr Leu Thr 
            100                 105                 110         


Asn Val Gln Asp Tyr Asn Gln Leu Thr Lys Met Leu Glu Glu Lys Phe 
        115                 120                 125             


Glu Asn Lys Asn Val Phe Tyr Ala Val Lys Leu Thr Gly Thr Phe Lys 
    130                 135                 140                 


Met Val Lys Ala Arg Thr Val Pro Lys Gln Thr Arg Pro Tyr Pro Gln 
145                 150                 155                 160 


Leu Thr Glu Val Thr Lys Lys Gln Ser Glu Phe Glu Phe Lys Asn Val 
                165                 170                 175     


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


Asn Val Pro Gly Phe His Leu His Phe Ile Thr Glu Asp Lys Thr Ser 
        195                 200                 205             


Gly Gly His Val Leu Asn Leu Gln Phe Asp Asn Ala Asn Leu Glu Ile 
    210                 215                 220                 


Ser Pro Ile His Glu Phe Asp Val Gln Leu Pro His Thr Asp Asp Phe 
225                 230                 235                 240 


Ala His Ser Asp Leu Thr Gln Val Thr Thr Ser Gln Val His Gln Ala 
                245                 250                 255     


Glu Ser Glu Arg Lys 
            260     


<210>  4
<211>  783
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  mature sequence of the aldB gene in plasmid RIHI-aldB

<400>  4
gctacaacgg ctactgtacc agcaccacct gccaagcagg aatccaaacc tgcggttgcc       60

gctaatccgg caccaaaaaa tgtactgttt caatactcaa cgatcaatgc actcatgctt      120

ggacagtttg aaggggactt gactttgaaa gacctgaagc tgcgaggcga tatggggctt      180

ggtaccatca atgatctcga tggagagatg attcagatgg gtacaaaatt ctaccagatc      240

gacagcaccg gaaaattatc ggagctgcca gaaagtgtga aaactccatt tgcggttact      300

acacatttcg agccgaaaga aaaaactaca ttaaccaatg tgcaagatta caatcaatta      360

acaaaaatgc ttgaggagaa atttgaaaac aagaacgtct tttatgccgt aaagctgacc      420

ggtactttta agatggtaaa ggctagaaca gttccaaaac aaaccagacc ttatccgcag      480

ctgactgaag taaccaaaaa acaatccgag tttgaattta aaaatgttaa gggaaccctg      540

attggcttct atacgccaaa ttatgcagca gccctgaatg ttcccggatt ccatctccac      600

ttcatcacag aggataaaac aagtggcgga cacgtattaa atctgcaatt tgacaacgcg      660

aatctggaaa tttctccgat ccatgagttt gatgttcaat tgccgcacac agatgatttt      720

gcccactctg atctgacaca agttactact agccaagtac accaagctga gtcagaaaga      780

aaa                                                                    783


<210>  5
<211>  290
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  aldB precursor protein expressed from plasmid RIHI-aldB

<400>  5

Val Arg Ser Lys Lys Leu Trp Ile Ser Leu Leu Phe Ala Leu Thr Leu 
1               5                   10                  15      


Ile Phe Thr Met Ala Phe Ser Asn Met Ser Ala Gln Ala Ala Thr Thr 
            20                  25                  30          


Ala Thr Val Pro Ala Pro Pro Ala Lys Gln Glu Ser Lys Pro Ala Val 
        35                  40                  45              


Ala Ala Asn Pro Ala Pro Lys Asn Val Leu Phe Gln Tyr Ser Thr Ile 
    50                  55                  60                  


Asn Ala Leu Met Leu Gly Gln Phe Glu Gly Asp Leu Thr Leu Lys Asp 
65                  70                  75                  80  


Leu Lys Leu Arg Gly Asp Met Gly Leu Gly Thr Ile Asn Asp Leu Asp 
                85                  90                  95      


Gly Glu Met Ile Gln Met Gly Thr Lys Phe Tyr Gln Ile Asp Ser Thr 
            100                 105                 110         


Gly Lys Leu Ser Glu Leu Pro Glu Ser Val Lys Thr Pro Phe Ala Val 
        115                 120                 125             


Thr Thr His Phe Glu Pro Lys Glu Lys Thr Thr Leu Thr Asn Val Gln 
    130                 135                 140                 


Asp Tyr Asn Gln Leu Thr Lys Met Leu Glu Glu Lys Phe Glu Asn Lys 
145                 150                 155                 160 


Asn Val Phe Tyr Ala Val Lys Leu Thr Gly Thr Phe Lys Met Val Lys 
                165                 170                 175     


Ala Arg Thr Val Pro Lys Gln Thr Arg Pro Tyr Pro Gln Leu Thr Glu 
            180                 185                 190         


Val Thr Lys Lys Gln Ser Glu Phe Glu Phe Lys Asn Val Lys Gly Thr 
        195                 200                 205             


Leu Ile Gly Phe Tyr Thr Pro Asn Tyr Ala Ala Ala Leu Asn Val Pro 
    210                 215                 220                 


Gly Phe His Leu His Phe Ile Thr Glu Asp Lys Thr Ser Gly Gly His 
225                 230                 235                 240 


Val Leu Asn Leu Gln Phe Asp Asn Ala Asn Leu Glu Ile Ser Pro Ile 
                245                 250                 255     


His Glu Phe Asp Val Gln Leu Pro His Thr Asp Asp Phe Ala His Ser 
            260                 265                 270         


Asp Leu Thr Gln Val Thr Thr Ser Gln Val His Gln Ala Glu Ser Glu 
        275                 280                 285             


Arg Lys 
    290 


<210>  6
<211>  858
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sequence encoding an acetolactate decarboxylase

<400>  6
atgaaaaaaa atatcatcac ttctatcaca tctctggctc tcgttgccgg gctgtctttg       60

actgcttttg cagctacaac ggctactgta ccagcaccac ctgccaagca ggaatccaaa      120

cctgtggttg ccgctaatcc ggcaccaaaa aatgtactgt ttcaatactc aacgatcaat      180

gcactcatgc ttggacagtt tgaaggggac ttgactttga aagacctgaa gctacgaggc      240

gatatggggc ttggtaccat caatgatctc gatggagaga tgattcagat gggtacaaaa      300

ttctaccaga tcgacagcac cggaaaatta tccgagctgc cagaaagtgt gaaaactcca      360

tttgcggtta ctacacattt cgagccgaaa gaaaaaacta cattaaccaa tgtgcaagat      420

tacaatcaat taacaaaaat gcttgaggag aaatttgaaa acaagaacgt cttttatgcc      480

gtaaagctga ccggtacctt taagatggta aaggctagaa cagttccaaa acaaaccaga      540

ccttatccgc agctgactga agtaaccaaa aaacaatccg agtttgaatt taaaaatgtt      600

aagggaaccc tgattggctt ctatacgcca aattatgcag cagccctgaa tgttcccgga      660

ttccatctcc acttcatcac agaggataaa acaagtggcg gacacgtatt aaatctgcaa      720

tttgacaacg cgaatctgga aatttctccg atccatgagt ttgatgtaca attgccgcac      780

acagatgatt ttgcccactc tgatctgaca caagttacta ctagccaagt acaccaagct      840

gagtcagaaa gaaaataa                                                    858


<210>  7
<211>  285
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  sequence of acetolactate decarboxylase (ALDC) precursor aldB

<400>  7

Met Lys Lys Asn Ile Ile Thr Ser Ile Thr Ser Leu Ala Leu Val Ala 
1               5                   10                  15      


Gly Leu Ser Leu Thr Ala Phe Ala Ala Thr Thr Ala Thr Val Pro Ala 
            20                  25                  30          


Pro Pro Ala Lys Gln Glu Ser Lys Pro Val Val Ala Ala Asn Pro Ala 
        35                  40                  45              


Pro Lys Asn Val Leu Phe Gln Tyr Ser Thr Ile Asn Ala Leu Met Leu 
    50                  55                  60                  


Gly Gln Phe Glu Gly Asp Leu Thr Leu Lys Asp Leu Lys Leu Arg Gly 
65                  70                  75                  80  


Asp Met Gly Leu Gly Thr Ile Asn Asp Leu Asp Gly Glu Met Ile Gln 
                85                  90                  95      


Met Gly Thr Lys Phe Tyr Gln Ile Asp Ser Thr Gly Lys Leu Ser Glu 
            100                 105                 110         


Leu Pro Glu Ser Val Lys Thr Pro Phe Ala Val Thr Thr His Phe Glu 
        115                 120                 125             


Pro Lys Glu Lys Thr Thr Leu Thr Asn Val Gln Asp Tyr Asn Gln Leu 
    130                 135                 140                 


Thr Lys Met Leu Glu Glu Lys Phe Glu Asn Lys Asn Val Phe Tyr Ala 
145                 150                 155                 160 


Val Lys Leu Thr Gly Thr Phe Lys Met Val Lys Ala Arg Thr Val Pro 
                165                 170                 175     


Lys Gln Thr Arg Pro Tyr Pro Gln Leu Thr Glu Val Thr Lys Lys Gln 
            180                 185                 190         


Ser Glu Phe Glu Phe Lys Asn Val Lys Gly Thr Leu Ile Gly Phe Tyr 
        195                 200                 205             


Thr Pro Asn Tyr Ala Ala Ala Leu Asn Val Pro Gly Phe His Leu His 
    210                 215                 220                 


Phe Ile Thr Glu Asp Lys Thr Ser Gly Gly His Val Leu Asn Leu Gln 
225                 230                 235                 240 


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


Gln Leu Pro His Thr Asp Asp Phe Ala His Ser Asp Leu Thr Gln Val 
            260                 265                 270         


Thr Thr Ser Gln Val His Gln Ala Glu Ser Glu Arg Lys 
        275                 280                 285 


<210>  8
<211>  261
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  predicted amino acid sequence of the mature acetolactate 
       decarboxylase (ALDC) aldB

<400>  8

Ala Thr Thr Ala Thr Val Pro Ala Pro Pro Ala Lys Gln Glu Ser Lys 
1               5                   10                  15      


Pro Val Val Ala Ala Asn Pro Ala Pro Lys Asn Val Leu Phe Gln Tyr 
            20                  25                  30          


Ser Thr Ile Asn Ala Leu Met Leu Gly Gln Phe Glu Gly Asp Leu Thr 
        35                  40                  45              


Leu Lys Asp Leu Lys Leu Arg Gly Asp Met Gly Leu Gly Thr Ile Asn 
    50                  55                  60                  


Asp Leu Asp Gly Glu Met Ile Gln Met Gly Thr Lys Phe Tyr Gln Ile 
65                  70                  75                  80  


Asp Ser Thr Gly Lys Leu Ser Glu Leu Pro Glu Ser Val Lys Thr Pro 
                85                  90                  95      


Phe Ala Val Thr Thr His Phe Glu Pro Lys Glu Lys Thr Thr Leu Thr 
            100                 105                 110         


Asn Val Gln Asp Tyr Asn Gln Leu Thr Lys Met Leu Glu Glu Lys Phe 
        115                 120                 125             


Glu Asn Lys Asn Val Phe Tyr Ala Val Lys Leu Thr Gly Thr Phe Lys 
    130                 135                 140                 


Met Val Lys Ala Arg Thr Val Pro Lys Gln Thr Arg Pro Tyr Pro Gln 
145                 150                 155                 160 


Leu Thr Glu Val Thr Lys Lys Gln Ser Glu Phe Glu Phe Lys Asn Val 
                165                 170                 175     


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


Asn Val Pro Gly Phe His Leu His Phe Ile Thr Glu Asp Lys Thr Ser 
        195                 200                 205             


Gly Gly His Val Leu Asn Leu Gln Phe Asp Asn Ala Asn Leu Glu Ile 
    210                 215                 220                 


Ser Pro Ile His Glu Phe Asp Val Gln Leu Pro His Thr Asp Asp Phe 
225                 230                 235                 240 


Ala His Ser Asp Leu Thr Gln Val Thr Thr Ser Gln Val His Gln Ala 
                245                 250                 255     


Glu Ser Glu Arg Lys 
            260     


