<110>    SAMYANG COPORATION

<120>    Glycosyltransferase and method of preparing steviol glycosides
         using the same

<130>    OPP20214954KR

<150>    KR 10-2020-0182517
<151>    2020-12-23

<160>    16

<170>    KoPatentIn 3.0

<210>    1
<211>    461
<212>    PRT
<213>    Artificial Sequence

<220>
<223>    TaUGT amino acid sequence


<400>    1
Met Asp Asp Gly Ser Ser Ser Ser Ser Ser Pro Leu Arg Val Val Ile
  1               5                  10                  15 

Cys Pro Trp Leu Ala Phe Gly His Leu Leu Pro Cys Leu Asp Ile Ala
             20                  25                  30 

Glu Arg Leu Ala Ser Arg Gly His Arg Val Ser Phe Val Ser Thr Pro
         35                  40                  45 

Arg Asn Ile Ala Arg Leu Pro Pro Val Arg Pro Ala Val Ala Pro Leu
     50                  55                  60 

Val Asp Tyr Val Ala Leu Pro Leu Pro Arg Val Asp Gly Leu Pro Glu
 65                  70                  75                  80 

Gly Ala Glu Ser Thr Asn Asp Val Pro His Asp Lys Phe Glu Leu Leu
                 85                  90                  95 

Arg Lys Ala Phe Asp Gly Leu Ala Ala Pro Phe Ser Glu Phe Leu His
            100                 105                 110 

Ala Ala Cys Ala Glu Gly Thr Gly Lys Arg Pro Asp Trp Leu Ile Val
        115                 120                 125 

Asp Ser Phe His His Trp Ala Ala Ala Ala Ala Val Glu Asn Lys Val
    130                 135                 140 

Pro Cys Val Met Leu Leu Leu Gly Ala Ala Asn Val Ile Ala Thr Trp
145                 150                 155                 160 

Ala Arg Gly Val Ser Glu His Ala Ala Ala Ala Val Gly Lys Glu Arg
                165                 170                 175 

Ser Ala Ala Glu Ala Pro Ser Phe Glu Thr Glu Arg Arg Lys Leu Met
            180                 185                 190 

Ile Thr Gln Asn Ala Ser Gly Met Thr Val Ala Glu Arg Tyr Phe Leu
        195                 200                 205 

Thr Leu Met Arg Ser Asn Leu Val Ala Ile Arg Ser Cys Ala Glu Trp
    210                 215                 220 

Glu Pro Glu Ser Val Ala Ala Leu Thr Thr Leu Ala Gly Lys Pro Val
225                 230                 235                 240 

Val Thr Leu Gly Leu Leu Pro Pro Ser Pro Glu Gly Gly Arg Gly Ile
                245                 250                 255 

Ser Lys Gln Asp Ala Ala Val Arg Trp Leu Asp Ala Gln Arg Asp Lys
            260                 265                 270 

Ser Val Val Tyr Val Ala Leu Gly Ser Glu Val Pro Leu Arg Val Glu
        275                 280                 285 

Gln Val His Glu Leu Ala Leu Gly Leu Glu Leu Ser Gly Ala Ser Phe
    290                 295                 300 

Leu Trp Ala Leu Arg Lys Pro Pro Gly Met Pro Asp Ala Ala Val Leu
305                 310                 315                 320 

Pro Pro Gly Phe Glu Glu Arg Thr Arg Gly Arg Gly Leu Val Val Thr
                325                 330                 335 

Gly Trp Val Pro Gln Ile Ser Val Leu Ala His Gly Ala Val Ala Ala
            340                 345                 350 

Phe Leu Thr His Cys Gly Trp Asn Ser Thr Ile Glu Gly Leu Leu Phe
        355                 360                 365 

Gly Gln Pro Leu Ile Met Leu Pro Ile Ser Ser Asp Gln Gly Pro Asn
    370                 375                 380 

Ala Arg Leu Met Glu Gly Arg Lys Val Gly Met Gln Val Pro Arg Asn
385                 390                 395                 400 

Glu Ser Asp Gly Ser Phe Thr Arg Glu Asp Val Ala Ala Thr Val Gln
                405                 410                 415 

Ala Val Ala Met Glu Glu Asp Gly Ser Arg Val Phe Thr Ala Asn Ala
            420                 425                 430 

Lys Thr Met Gln Glu Ile Val Ala Asp Ser Ala Cys His Glu Arg Cys
        435                 440                 445 

Ile Asp Gly Phe Ile Gln Gln Leu Arg Ser Tyr Lys Glu
    450                 455                 460 




<210>    2
<211>    1383
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    TaUGT nucleotide sequence


<400>    2
atggacgacg ggtcttctag ctctagcagt ccgctgcgtg ttgttatttg cccgtggctg         60

gcttttggac acctgcttcc atgcttagac attgcggaaa gattagccag ccgtggacac        120

agagtatcct tcgtaagtac gccgcgtaat atagcacgtt tacccccggt cagaccggca        180

gttgcgccgc tggtcgatta cgtagcgcta cctctgccca gagtggacgg acttccggaa        240

ggggctgaat ctacaaacga cgtgccccat gataagttcg agcttttacg taaggctttc        300

gacggtttgg cagcgccctt cagcgaattt ctacacgcag cgtgcgcgga gggcacaggc        360

aaaagaccag attggcttat tgtggattcc ttccaccatt gggctgcggc tgcggcagtg        420

gaaaataaag tcccgtgcgt catgttactt ttaggtgcag ccaacgtgat cgccacttgg        480

gcacgtggcg tgtcagagca tgcggccgcc gcggttggaa aagaacgtag tgccgcagaa        540

gctccatcat tcgaaaccga acgtcgtaaa ctgatgataa ctcagaatgc atcaggtatg        600

accgtagcag agcgttattt cctgacactt atgagatcaa acttggttgc cattcgttcc        660

tgtgcagaat gggagcctga atctgtggcg gcactaacca ctctggccgg gaagcccgta        720

gtgaccttag gtttgcttcc accgtctcct gagggaggaa ggggtatttc caagcaagac        780

gctgccgtta ggtggctgga cgctcagagg gataagtccg ttgtatacgt ggcgttaggg        840

tctgaagttc ccttgagagt ggagcaggta catgagcttg cgcttggact agagttgtca        900

ggtgctagct tcctgtgggc gttacgtaag ccacccggca tgcctgacgc cgcagtcttg        960

ccgccgggtt ttgaagagag aaccagggga cgtggtctgg ttgtgactgg ctgggtacca       1020

caaatcagcg tcttagcaca tggcgccgtt gccgcttttc tgacacattg cggctggaat       1080

agtacaatcg aaggcttgct gttcggacaa ccgttaatca tgctgccaat cagtagcgat       1140

cagggcccga atgcccgtct tatggaggga agaaaagttg gaatgcaggt gcctcgtaat       1200

gagtccgatg gatcattcac aagagaggat gtcgctgcca cagtacaagc agtagcgatg       1260

gaggaggatg ggagccgtgt ttttacggct aatgcaaaga ccatgcaaga aatagttgct       1320

gactccgcgt gtcacgaaag gtgcatcgat ggatttatcc aacagctaag gtcctataaa       1380

gaa                                                                     1383


<210>    3
<211>    462
<212>    PRT
<213>    Artificial Sequence

<220>
<223>    EUGT11 amino acid sequence


<400>    3
Met Asp Ser Gly Tyr Ser Ser Ser Tyr Ala Ala Ala Ala Gly Met His
  1               5                  10                  15 

Val Val Ile Cys Pro Trp Leu Ala Phe Gly His Leu Leu Pro Cys Leu
             20                  25                  30 

Asp Leu Ala Gln Arg Leu Ala Ser Arg Gly His Arg Val Ser Phe Val
         35                  40                  45 

Ser Thr Pro Arg Asn Ile Ser Arg Leu Pro Pro Val Arg Pro Ala Leu
     50                  55                  60 

Ala Pro Leu Val Ala Phe Val Ala Leu Pro Leu Pro Arg Val Glu Gly
 65                  70                  75                  80 

Leu Pro Asp Gly Ala Glu Ser Thr Asn Asp Val Pro His Asp Arg Pro
                 85                  90                  95 

Asp Met Val Glu Leu His Arg Arg Ala Phe Asp Gly Leu Ala Ala Pro
            100                 105                 110 

Phe Ser Glu Phe Leu Gly Thr Ala Cys Ala Asp Trp Val Ile Val Asp
        115                 120                 125 

Val Phe His His Trp Ala Ala Ala Ala Ala Leu Glu His Lys Val Pro
    130                 135                 140 

Cys Ala Met Met Leu Leu Gly Ser Ala His Met Ile Ala Ser Ile Ala
145                 150                 155                 160 

Asp Arg Arg Leu Glu Arg Ala Glu Thr Glu Ser Pro Ala Ala Ala Gly
                165                 170                 175 

Gln Gly Arg Pro Ala Ala Ala Pro Thr Phe Glu Val Ala Arg Met Lys
            180                 185                 190 

Leu Ile Arg Thr Lys Gly Ser Ser Gly Met Ser Leu Ala Glu Arg Phe
        195                 200                 205 

Ser Leu Thr Leu Ser Arg Ser Ser Leu Val Val Gly Arg Ser Cys Val
    210                 215                 220 

Glu Phe Glu Pro Glu Thr Val Pro Leu Leu Ser Thr Leu Arg Gly Lys
225                 230                 235                 240 

Pro Ile Thr Phe Leu Gly Leu Met Pro Pro Leu His Glu Gly Arg Arg
                245                 250                 255 

Glu Asp Gly Glu Asp Ala Thr Val Arg Trp Leu Asp Ala Gln Pro Ala
            260                 265                 270 

Lys Ser Val Val Tyr Val Ala Leu Gly Ser Glu Val Pro Leu Gly Val
        275                 280                 285 

Glu Lys Val His Glu Leu Ala Leu Gly Leu Glu Leu Ala Gly Thr Arg
    290                 295                 300 

Phe Leu Trp Ala Leu Arg Lys Pro Thr Gly Val Ser Asp Ala Asp Leu
305                 310                 315                 320 

Leu Pro Ala Gly Phe Glu Glu Arg Thr Arg Gly Arg Gly Val Val Ala
                325                 330                 335 

Thr Arg Trp Val Pro Gln Met Ser Ile Leu Ala His Ala Ala Val Gly
            340                 345                 350 

Ala Phe Leu Thr His Cys Gly Trp Asn Ser Thr Ile Glu Gly Leu Met
        355                 360                 365 

Phe Gly His Pro Leu Ile Met Leu Pro Ile Phe Gly Asp Gln Gly Pro
    370                 375                 380 

Asn Ala Arg Leu Ile Glu Ala Lys Asn Ala Gly Leu Gln Val Ala Arg
385                 390                 395                 400 

Asn Asp Gly Asp Gly Ser Phe Asp Arg Glu Gly Val Ala Ala Ala Ile
                405                 410                 415 

Arg Ala Val Ala Val Glu Glu Glu Ser Ser Lys Val Phe Gln Ala Lys
            420                 425                 430 

Ala Lys Lys Leu Gln Glu Ile Val Ala Asp Met Ala Cys His Glu Arg
        435                 440                 445 

Tyr Ile Asp Gly Phe Ile Gln Gln Leu Arg Ser Tyr Lys Asp
    450                 455                 460 




<210>    4
<211>    1386
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    EUGT11 nucleotide sequence


<400>    4
atggactccg gctacagttc atcatatgct gcagccgctg ggatgcacgt tgtgatttgt         60

ccgtggctag cattcggtca tctgttgccg tgtttggacc tggctcagag gctagcttcc        120

agaggacatc gtgtttcctt cgtttctacg cctaggaata tcagtagact accaccggta        180

agacctgctt tagctccact tgtggcgttc gtggccctac cgttaccccg tgtagaagga        240

ttgcctgacg gagcagagtc tacgaatgac gtacctcacg acagacccga tatggttgaa        300

cttcacagaa gagcgttcga cggtcttgct gcaccgtttt ccgaattcct tgggacggcc        360

tgtgcggact gggtcatagt ggacgtattc catcactggg ctgccgcagc tgccctagaa        420

cataaggttc cgtgtgccat gatgttgtta ggttctgctc atatgatcgc gtctattgca        480

gaccgtaggt tggagagggc agaaacggaa agtccagccg ccgccggaca ggggaggccc        540

gctgcggcgc caacctttga ggtggctcgt atgaaactaa ttaggactaa aggttcaagc        600

ggtatgtcac tggcggaaag gtttagtctt actttgtcca ggtcttcatt ggtcgtcggt        660

cgttcctgcg ttgagtttga acccgaaacc gttccccttc tttccacgtt acgtggaaaa        720

ccgattactt ttcttggtct tatgccgcca ctgcacgagg gtagaaggga agatggtgaa        780

gacgctacag ttaggtggct agacgcgcaa ccggctaaga gcgtcgtcta tgtcgcactt        840

ggctcagagg tgcccttggg ggtcgagaag gtccatgagt tggcgctggg gttggagttg        900

gcgggtacaa ggtttctttg ggcccttcgt aaaccgacgg gcgtatcaga tgcagaccta        960

cttccggctg gtttcgagga gcgtactagg ggaaggggcg ttgtggccac gagatgggtg       1020

ccacaaatga gcatcctagc tcatgccgca gtcggcgcat tcttaacgca ttgtggctgg       1080

aattcaacca ttgaagggct aatgttcggt catccactga taatgcttcc tatctttgga       1140

gaccaaggtc ccaatgcgcg tttgatcgaa gccaagaatg ctggccttca agtcgcccgt       1200

aatgatggcg atggttcatt cgacagagag ggagtggcgg cagcgatcag agcggttgca       1260

gtggaagagg agtccagtaa ggtgtttcag gctaaggcca agaaattaca ggagatagtg       1320

gctgatatgg cttgccacga gagatatatc gacggcttta tacaacagct acgttcttat       1380

aaggac                                                                  1386


<210>    5
<211>    463
<212>    PRT
<213>    Artificial Sequence

<220>
<223>    HvUGT amino acid sequence


<400>    5
Met Asp Gly Asp Gly Asn Ser Ser Ser Ser Ser Ser Pro Leu His Val
  1               5                  10                  15 

Val Ile Cys Pro Trp Leu Ala Leu Gly His Leu Leu Pro Cys Leu Asp
             20                  25                  30 

Ile Ala Glu Arg Leu Ala Ser Arg Gly His Arg Val Ser Phe Val Ser
         35                  40                  45 

Thr Pro Arg Asn Ile Ala Arg Leu Pro Pro Leu Arg Pro Ala Val Ala
     50                  55                  60 

Pro Leu Val Glu Phe Val Ala Leu Pro Leu Pro His Val Asp Gly Leu
 65                  70                  75                  80 

Pro Glu Gly Ala Glu Ser Thr Asn Asp Val Pro Tyr Asp Lys Phe Glu
                 85                  90                  95 

Leu His Arg Lys Ala Phe Asp Gly Leu Ala Ala Pro Phe Ser Glu Phe
            100                 105                 110 

Leu Arg Ala Ala Cys Ala Glu Gly Ala Gly Ser Arg Pro Asp Trp Leu
        115                 120                 125 

Ile Val Asp Thr Phe His His Trp Ala Ala Ala Ala Ala Val Glu Asn
    130                 135                 140 

Lys Val Pro Cys Val Met Leu Leu Leu Gly Ala Ala Thr Val Ile Ala
145                 150                 155                 160 

Gly Phe Ala Arg Gly Val Ser Glu His Ala Ala Ala Ala Val Gly Lys
                165                 170                 175 

Glu Arg Pro Ala Ala Glu Ala Pro Ser Phe Glu Thr Glu Arg Arg Lys
            180                 185                 190 

Leu Met Thr Thr Gln Asn Ala Ser Gly Met Thr Val Ala Glu Arg Tyr
        195                 200                 205 

Phe Leu Thr Leu Met Arg Ser Asp Leu Val Ala Ile Arg Ser Cys Ala
    210                 215                 220 

Glu Trp Glu Pro Glu Ser Val Ala Ala Leu Thr Thr Leu Ala Gly Lys
225                 230                 235                 240 

Pro Val Val Pro Leu Gly Leu Leu Pro Pro Ser Pro Glu Gly Gly Arg
                245                 250                 255 

Gly Val Ser Lys Glu Asp Ala Ala Val Arg Trp Leu Asp Ala Gln Pro
            260                 265                 270 

Ala Lys Ser Val Val Tyr Val Ala Leu Gly Ser Glu Val Pro Leu Arg
        275                 280                 285 

Ala Glu Gln Val His Glu Leu Ala Leu Gly Leu Glu Leu Ser Gly Ala
    290                 295                 300 

Arg Phe Leu Trp Ala Leu Arg Lys Pro Thr Asp Ala Pro Asp Ala Ala
305                 310                 315                 320 

Val Leu Pro Pro Gly Phe Glu Glu Arg Thr Arg Gly Arg Gly Leu Val
                325                 330                 335 

Val Thr Gly Trp Val Pro Gln Ile Gly Val Leu Ala His Gly Ala Val
            340                 345                 350 

Ala Ala Phe Leu Thr His Cys Gly Trp Asn Ser Thr Ile Glu Gly Leu
        355                 360                 365 

Leu Phe Gly His Pro Leu Ile Met Leu Pro Ile Ser Ser Asp Gln Gly
    370                 375                 380 

Pro Asn Ala Arg Leu Met Glu Gly Arg Lys Val Gly Met Gln Val Pro
385                 390                 395                 400 

Arg Asp Glu Ser Asp Gly Ser Phe Arg Arg Glu Asp Val Ala Ala Thr
                405                 410                 415 

Val Arg Ala Val Ala Val Glu Glu Asp Gly Arg Arg Val Phe Thr Ala
            420                 425                 430 

Asn Ala Lys Lys Met Gln Glu Ile Val Ala Asp Gly Ala Cys His Glu
        435                 440                 445 

Arg Cys Ile Asp Gly Phe Ile Gln Gln Leu Arg Ser Tyr Lys Ala
    450                 455                 460 




<210>    6
<211>    1389
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    HvUGT nucleotide sequence


<400>    6
atggacggcg acggaaacag ttcatcaagt tctagtcctc tgcatgttgt tatatgtccc         60

tggctagctc ttgggcatct tcttccttgt ttggacattg cggaaagact ggcctcaaga        120

ggacacaggg tcagcttcgt atccacgcca agaaatattg cccgtctacc cccgcttagg        180

ccagcggtgg ctcccttagt agagttcgtt gctctacctc tgcctcacgt ggacgggtta        240

cctgaagggg ctgaaagtac aaacgatgta ccctatgaca agtttgaact tcaccgtaag        300

gcgttcgacg ggctggccgc tccttttagc gagttcttga gagctgcttg tgccgagggg        360

gctggttccc gtcctgattg gctaatcgtg gatacattcc atcactgggc agcggcagcc        420

gccgttgaaa ataaagtgcc atgcgtgatg ttactgcttg gtgcagccac cgttattgcc        480

gggttcgcaa gaggggtcag tgagcacgcc gccgctgcag tcggtaaaga gagaccggcc        540

gctgaagccc cgagttttga gacggagaga cgtaaactta tgaccactca gaatgcaagt        600

ggtatgacgg ttgcagaaag gtatttccta accttaatgc gtagtgactt ggtcgcaata        660

cgtagctgcg ccgagtggga gcccgagtct gtcgcggcgt tgacaacctt agcaggcaag        720

cccgtcgttc ctttaggctt gctgccaccg tctccagagg gggggagagg cgtttccaag        780

gaggacgccg cggttcgttg gttggatgcg caacccgcga agagcgtcgt gtatgtcgcg        840

ttaggcagtg aggttccatt gagagctgaa caagtccacg agttggcgct tggtttagaa        900

ttatctggcg ctaggttttt atgggcgcta aggaaaccca ccgacgcgcc agatgccgcg        960

gtattgccgc cagggtttga ggaaaggact cgtggcagag ggcttgtcgt gaccggttgg       1020

gtgcctcaaa tcggcgttct tgctcatggt gccgtagcag cattcttgac tcattgtggc       1080

tggaattcaa ccatcgaggg gctgctgttt ggtcacccgt tgatcatgct gccaataagc       1140

tccgaccaag gacccaatgc tcgtttgatg gaaggcagaa aagtggggat gcaggtgccc       1200

agagacgaaa gcgatgggag ttttaggaga gaagatgtgg cagctactgt tcgtgctgta       1260

gcggtggagg aagatggcag aagggttttt acagctaatg cgaagaagat gcaagaaata       1320

gtagccgacg gggcttgtca cgagagatgc atcgatggat ttatccaaca actgcgttcc       1380

tacaaagct                                                               1389


<210>    7
<211>    31
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Forward primer for CYC1 terminator


<400>    7
tgattgtcga tatcatgtaa ttagttatgt c                                        31


<210>    8
<211>    28
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Reverse primer for GAL10 promoter


<400>    8
catcaattct tacttttttt ttggatgg                                            28


<210>    9
<211>    44
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Forward primer for connecting TaUGT and GAL10 promoter


<400>    9
catccaaaaa aaaagtaaga attgatggac gacgggtctt ctag                          44


<210>    10
<211>    48
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Reverse primer for connecting TaUGT and CYC1 terminator


<400>    10
ctaattacat gatatcgaca atcattcttt ataggacctt agctgttg                      48


<210>    11
<211>    44
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Forward primer for connecting EUGT11 and GAL10 promoter


<400>    11
catccaaaaa aaaagtaaga attgatggac tccggctaca gttc                          44


<210>    12
<211>    45
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Reverse primer for connecting EUGT11 and CYC1 terminator


<400>    12
ctaattacat gatatcgaca attagtcctt ataagaacgt agctg                         45


<210>    13
<211>    42
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Forward primer for connecting HvUGT and GAL10 promoter


<400>    13
catccaaaaa aaaagtaaga attgatggac ggcgacggaa ac                            42


<210>    14
<211>    45
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Reverse primer for connecting HvUGT and CYC1 terminator


<400>    14
ctaattacat gatatcgaca atcaagcttt gtaggaacgc agttg                         45


<210>    15
<211>    18
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Forward primer for GAL10 promoter


<400>    15
ggatccatcg cttcgctg                                                       18


<210>    16
<211>    20
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Reverse primer for CYC1 terminator


<400>    16
gcaaattaaa gccttcgagc                                                     20
