<110>    CJ CheilJedang Corporation

<120>    Novel major facilitator superfamily permease variant and a method
         for producing L-lysine using the same

<130>    OPP20212333KR

<150>    KR 10-2021-0047409
<151>    2021-04-12

<160>    10

<170>    KoPatentIn 3.0

<210>    1
<211>    481
<212>    PRT
<213>    Artificial Sequence

<220>
<223>    (H377Q)_major facilitator superfamily permease


<400>    1
Met Asp Ser Gln Ile Asn Thr Gln Thr Ser Pro Ala Ala Ala Lys Leu
  1               5                  10                  15 

Pro Arg Glu Val Val Val Val Leu Ser Ile Leu Val Val Ser Ala Met
             20                  25                  30 

Ile Met Ile Leu Asn Glu Thr Ile Leu Ser Val Ala Leu Pro Ser Ile
         35                  40                  45 

Met Glu Asp Phe Ser Val Pro Glu Thr Thr Ala Gln Trp Leu Thr Thr
     50                  55                  60 

Gly Phe Met Leu Thr Met Ala Val Val Ile Pro Thr Thr Gly Tyr Leu
 65                  70                  75                  80 

Leu Asp Arg Phe Ser Thr Lys Thr Ile Phe Val Thr Ala Leu Leu Phe
                 85                  90                  95 

Phe Thr Val Gly Thr Leu Thr Ala Ala Leu Ala Pro Thr Phe Ala Val
            100                 105                 110 

Leu Leu Gly Ala Arg Ile Val Gln Ala Val Gly Thr Ala Leu Val Met
        115                 120                 125 

Pro Leu Leu Met Thr Val Thr Leu Thr Val Val Pro Ala Glu Arg Arg
    130                 135                 140 

Gly Ser Met Met Gly Ile Ile Ser Ile Val Ile Ser Val Ala Pro Ala
145                 150                 155                 160 

Leu Gly Pro Thr Leu Ser Gly Val Ile Leu Asn Ser Leu Thr Trp His
                165                 170                 175 

Trp Leu Phe Trp Met Met Leu Pro Ile Val Val Ile Ala Leu Val Ile
            180                 185                 190 

Gly Phe Phe Leu Ile Lys Asn Ile Gly Glu Thr Lys Ile Thr Pro Leu
        195                 200                 205 

Asp Val Leu Ser Val Ile Leu Ser Val Phe Ala Phe Gly Gly Leu Val
    210                 215                 220 

Tyr Gly Phe Ser Ser Phe Gly Ala Ile Leu Glu Gly Glu Gly Thr Val
225                 230                 235                 240 

Gly Ile Phe Ala Ile Val Val Gly Ala Ile Ala Leu Leu Ile Phe Ala
                245                 250                 255 

Leu Arg Gln His Gln Leu Gly Lys Gln Asp Lys Ala Leu Met Asp Leu
            260                 265                 270 

Arg Ala Phe Lys Val Arg Asn Phe Ser Phe Ser Leu Thr Thr Ile Leu
        275                 280                 285 

Leu Ala Phe Gly Ala Met Leu Gly Thr Val Met Val Leu Pro Ile Tyr
    290                 295                 300 

Leu Gln Thr Ser Leu Gly Val Thr Ala Leu Val Thr Gly Leu Val Val
305                 310                 315                 320 

Met Pro Gly Gly Leu Leu Gln Gly Leu Ile Ser Pro Phe Ile Gly Arg
                325                 330                 335 

Phe Tyr Asp Lys Val Gly Pro Arg Pro Leu Leu Ile Pro Gly Ala Ile
            340                 345                 350 

Ala Leu Ala Ile Ala Ala Ser Ser Met Thr Phe Leu Asn Glu Asn Ser
        355                 360                 365 

Pro Val Trp Met Val Val Val Met Gln Val Val Phe Ser Ile Gly Met
    370                 375                 380 

Cys Leu Met Met Thr Pro Leu Met Thr Thr Ala Leu Gly Ala Leu Pro
385                 390                 395                 400 

Lys His Leu Tyr Gly His Gly Ser Ala Ile Leu Asn Thr Phe Gln Gln
                405                 410                 415 

Leu Ala Gly Ala Ala Gly Thr Ala Ile Met Ile Ala Ala Leu Ser Phe
            420                 425                 430 

Gly Thr Ser Ile Ala Ala Ser Ser Gly Ser Ala His Ala Glu Ala Val
        435                 440                 445 

Ala Ala Gly Thr Lys Val Ala Phe Ile Ala Gly Ala Ile Ile Ala Val
    450                 455                 460 

Ile Ala Leu Val Val Ser Leu Phe Val Thr Arg Val Glu Glu Glu Ala
465                 470                 475                 480 

His
   


<210>    2
<211>    1446
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    (H377Q)_NCgl2592


<400>    2
atggattccc aaattaatac tcagacctct ccggcagctg cgaagctgcc tagggaggtc         60

gttgttgttc tttcgatcct cgtggtttcc gcgatgatca tgattcttaa tgaaaccatt        120

ctgtcggttg cgttgccttc catcatggaa gatttctccg tgcctgaaac tactgcacag        180

tggttgacca ctggctttat gttgacgatg gcagtggtga ttccaactac tggttatctg        240

cttgatcgtt tttccactaa gacgatcttt gttactgcgt tgttgttctt tacggttggt        300

acgttgactg cggcgttggc tccaacgttt gcggtgctgc ttggtgctcg tatcgttcag        360

gcggttggta ctgcgctggt gatgcctttg ctgatgacgg ttacgttgac ggtggttcct        420

gcggagcgtc gtggttcgat gatgggcatt atttccatcg tgatttctgt tgcgccggcg        480

cttggtccta cgttgtctgg tgtcattctt aactctttga cctggcactg gttgttttgg        540

atgatgcttc cgatcgttgt tatcgctttg gtaattggtt tcttcttgat caaaaatatc        600

ggcgaaacca agatcacccc actggatgtt ctgtctgtca tcctttcggt gtttgccttc        660

ggtggtttgg tgtacggctt cagttccttc ggagcaatcc tggagggcga aggcaccgta        720

ggtatcttcg cgatcgtcgt tggcgccatc gcactcctca tctttgcttt gcgacagcac        780

caactcggca agcaagacaa agcactgatg gatctccgag ccttcaaggt gaggaacttc        840

agcttctcct tgaccaccat ccttttggcg ttcggcgcga tgctcggaac cgtcatggtt        900

ttgccaatct acctgcagac ttccctcgga gttactgctt tggtgaccgg tttggttgtt        960

atgcccggcg gcctcctcca gggtctgatc agcccattca tcggacgttt ctacgacaag       1020

gtcggtccac gtccgctgct gattcccgga gcaattgcgc tggctatcgc ggcatcctcg       1080

atgactttcc tcaatgagaa ttcacccgtg tggatggtcg tggtcatgca agttgtgttc       1140

agcatcggca tgtgtttgat gatgacccct ctcatgacca ccgctctcgg cgcccttccg       1200

aagcacctct atggtcacgg ctccgcaatt ttgaacacgt tccaacagct cgcaggcgca       1260

gccggaacag cgatcatgat tgcagcactt tccttcggca cttccattgc agcgtcttcg       1320

ggatctgcgc atgctgaagc tgttgccgct ggtaccaagg ttgcgttcat cgcaggcgca       1380

atcatcgcgg tgatcgcttt ggttgtttcc ctcttcgtca ctcgcgtcga ggaagaagct       1440

cactaa                                                                  1446


<210>    3
<211>    481
<212>    PRT
<213>    Artificial Sequence

<220>
<223>    (WT)_major facilitator superfamily permease


<400>    3
Met Asp Ser Gln Ile Asn Thr Gln Thr Ser Pro Ala Ala Ala Lys Leu
  1               5                  10                  15 

Pro Arg Glu Val Val Val Val Leu Ser Ile Leu Val Val Ser Ala Met
             20                  25                  30 

Ile Met Ile Leu Asn Glu Thr Ile Leu Ser Val Ala Leu Pro Ser Ile
         35                  40                  45 

Met Glu Asp Phe Ser Val Pro Glu Thr Thr Ala Gln Trp Leu Thr Thr
     50                  55                  60 

Gly Phe Met Leu Thr Met Ala Val Val Ile Pro Thr Thr Gly Tyr Leu
 65                  70                  75                  80 

Leu Asp Arg Phe Ser Thr Lys Thr Ile Phe Val Thr Ala Leu Leu Phe
                 85                  90                  95 

Phe Thr Val Gly Thr Leu Thr Ala Ala Leu Ala Pro Thr Phe Ala Val
            100                 105                 110 

Leu Leu Gly Ala Arg Ile Val Gln Ala Val Gly Thr Ala Leu Val Met
        115                 120                 125 

Pro Leu Leu Met Thr Val Thr Leu Thr Val Val Pro Ala Glu Arg Arg
    130                 135                 140 

Gly Ser Met Met Gly Ile Ile Ser Ile Val Ile Ser Val Ala Pro Ala
145                 150                 155                 160 

Leu Gly Pro Thr Leu Ser Gly Val Ile Leu Asn Ser Leu Thr Trp His
                165                 170                 175 

Trp Leu Phe Trp Met Met Leu Pro Ile Val Val Ile Ala Leu Val Ile
            180                 185                 190 

Gly Phe Phe Leu Ile Lys Asn Ile Gly Glu Thr Lys Ile Thr Pro Leu
        195                 200                 205 

Asp Val Leu Ser Val Ile Leu Ser Val Phe Ala Phe Gly Gly Leu Val
    210                 215                 220 

Tyr Gly Phe Ser Ser Phe Gly Ala Ile Leu Glu Gly Glu Gly Thr Val
225                 230                 235                 240 

Gly Ile Phe Ala Ile Val Val Gly Ala Ile Ala Leu Leu Ile Phe Ala
                245                 250                 255 

Leu Arg Gln His Gln Leu Gly Lys Gln Asp Lys Ala Leu Met Asp Leu
            260                 265                 270 

Arg Ala Phe Lys Val Arg Asn Phe Ser Phe Ser Leu Thr Thr Ile Leu
        275                 280                 285 

Leu Ala Phe Gly Ala Met Leu Gly Thr Val Met Val Leu Pro Ile Tyr
    290                 295                 300 

Leu Gln Thr Ser Leu Gly Val Thr Ala Leu Val Thr Gly Leu Val Val
305                 310                 315                 320 

Met Pro Gly Gly Leu Leu Gln Gly Leu Ile Ser Pro Phe Ile Gly Arg
                325                 330                 335 

Phe Tyr Asp Lys Val Gly Pro Arg Pro Leu Leu Ile Pro Gly Ala Ile
            340                 345                 350 

Ala Leu Ala Ile Ala Ala Ser Ser Met Thr Phe Leu Asn Glu Asn Ser
        355                 360                 365 

Pro Val Trp Met Val Val Val Met His Val Val Phe Ser Ile Gly Met
    370                 375                 380 

Cys Leu Met Met Thr Pro Leu Met Thr Thr Ala Leu Gly Ala Leu Pro
385                 390                 395                 400 

Lys His Leu Tyr Gly His Gly Ser Ala Ile Leu Asn Thr Phe Gln Gln
                405                 410                 415 

Leu Ala Gly Ala Ala Gly Thr Ala Ile Met Ile Ala Ala Leu Ser Phe
            420                 425                 430 

Gly Thr Ser Ile Ala Ala Ser Ser Gly Ser Ala His Ala Glu Ala Val
        435                 440                 445 

Ala Ala Gly Thr Lys Val Ala Phe Ile Ala Gly Ala Ile Ile Ala Val
    450                 455                 460 

Ile Ala Leu Val Val Ser Leu Phe Val Thr Arg Val Glu Glu Glu Ala
465                 470                 475                 480 

His
   


<210>    4
<211>    1446
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    (WT)_NCgl2592


<400>    4
atggattccc aaattaatac tcagacctct ccggcagctg cgaagctgcc tagggaggtc         60

gttgttgttc tttcgatcct cgtggtttcc gcgatgatca tgattcttaa tgaaaccatt        120

ctgtcggttg cgttgccttc catcatggaa gatttctccg tgcctgaaac tactgcacag        180

tggttgacca ctggctttat gttgacgatg gcagtggtga ttccaactac tggttatctg        240

cttgatcgtt tttccactaa gacgatcttt gttactgcgt tgttgttctt tacggttggt        300

acgttgactg cggcgttggc tccaacgttt gcggtgctgc ttggtgctcg tatcgttcag        360

gcggttggta ctgcgctggt gatgcctttg ctgatgacgg ttacgttgac ggtggttcct        420

gcggagcgtc gtggttcgat gatgggcatt atttccatcg tgatttctgt tgcgccggcg        480

cttggtccta cgttgtctgg tgtcattctt aactctttga cctggcactg gttgttttgg        540

atgatgcttc cgatcgttgt tatcgctttg gtaattggtt tcttcttgat caaaaatatc        600

ggcgaaacca agatcacccc actggatgtt ctgtctgtca tcctttcggt gtttgccttc        660

ggtggtttgg tgtacggctt cagttccttc ggagcaatcc tggagggcga aggcaccgta        720

ggtatcttcg cgatcgtcgt tggcgccatc gcactcctca tctttgcttt gcgacagcac        780

caactcggca agcaagacaa agcactgatg gatctccgag ccttcaaggt gaggaacttc        840

agcttctcct tgaccaccat ccttttggcg ttcggcgcga tgctcggaac cgtcatggtt        900

ttgccaatct acctgcagac ttccctcgga gttactgctt tggtgaccgg tttggttgtt        960

atgcccggcg gcctcctcca gggtctgatc agcccattca tcggacgttt ctacgacaag       1020

gtcggtccac gtccgctgct gattcccgga gcaattgcgc tggctatcgc ggcatcctcg       1080

atgactttcc tcaatgagaa ttcacccgtg tggatggtcg tggtcatgca cgttgtgttc       1140

agcatcggca tgtgtttgat gatgacccct ctcatgacca ccgctctcgg cgcccttccg       1200

aagcacctct atggtcacgg ctccgcaatt ttgaacacgt tccaacagct cgcaggcgca       1260

gccggaacag cgatcatgat tgcagcactt tccttcggca cttccattgc agcgtcttcg       1320

ggatctgcgc atgctgaagc tgttgccgct ggtaccaagg ttgcgttcat cgcaggcgca       1380

atcatcgcgg tgatcgcttt ggttgtttcc ctcttcgtca ctcgcgtcga ggaagaagct       1440

cactaa                                                                  1446


<210>    5
<211>    38
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    NCgl2592_1F


<400>    5
tgaattcgag ctcggtaccc cggtggtttg gtgtacgg                                 38


<210>    6
<211>    33
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    NCgl2592_2R


<400>    6
cgatgctgaa cacaacttgc atgaccacga cca                                      33


<210>    7
<211>    33
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    NCgl2592_3F


<400>    7
tggtcgtggt catgcaagtt gtgttcagca tcg                                      33


<210>    8
<211>    38
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    NCgl2592_4R


<400>    8
gtcgactcta gaggatcccc cgcttaaggt gaggtggc                                 38


<210>    9
<211>    18
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    NCgl2592_5F


<400>    9
ggatgatgct tccgatcg                                                       18


<210>    10
<211>    18
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    NCgl2592_6R


<400>    10
gccgacctac caacaagc                                                       18
