WO2022149865A2 - GlxR 단백질 변이체 또는 이를 이용한 쓰레오닌 생산방법 - Google Patents
GlxR 단백질 변이체 또는 이를 이용한 쓰레오닌 생산방법 Download PDFInfo
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- WO2022149865A2 WO2022149865A2 PCT/KR2022/000204 KR2022000204W WO2022149865A2 WO 2022149865 A2 WO2022149865 A2 WO 2022149865A2 KR 2022000204 W KR2022000204 W KR 2022000204W WO 2022149865 A2 WO2022149865 A2 WO 2022149865A2
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- glxr
- seq
- amino acid
- threonine
- microorganism
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/34—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Corynebacterium (G)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N15/77—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Corynebacterium; for Brevibacterium
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/08—Lysine; Diaminopimelic acid; Threonine; Valine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/15—Corynebacterium
Abstract
Description
명칭 | 서열 | 서열번호 |
GlxR_amino acid | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGEPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 1 |
glxR_nucleic acid | GTGGAAGGTGTACAGGAGATCCTGTCGCGCGCCGGAATTTTTCAAGGCGTTGACCCAACGGCAGTCAATAACCTCATCCAGGATATGGAGACCGTTCGCTTCCCACGCGGAGCAACCATCTTCGACGAGGGCGAGCCAGGTGACCGCCTTTACATCATCACCTCCGGCAAAGTGAAGCTTGCGCGCCACGCACCGGACGGCCGCGAAAACCTGCTGACCATCATGGGTCCTTCCGACATGTTCGGTGAGCTCTCCATCTTCGACCCAGGCCCACGCACCTCCTCTGCAGTGTGTGTCACCGAAGTTCATGCAGCAACCATGAACTCTGACATGCTGCGCAACTGGGTAGCTGACCACCCAGCTATCGCTGAGCAGCTCCTGCGCGTTCTGGCTCGTCGTCTGCGTCGCACCAACGCTTCCCTGGCTGACCTCATCTTCACCGACGTCCCAGGCCGCGTTGCTAAGACCCTTCTGCAGCTGGCTAACCGCTTCGGCACCCAAGAAGCTGGCGCGCTGCGCGTGAACCACGACCTCACTCAGGAAGAAATCGCACAGCTCGTCGGTGCTTCCCGTGAAACTGTGAATAAGGCTCTTGCAACGTTCGCACACCGTGGCTGGATTCGCCTCGAGGGCAAGTCCGTCCTCATTGTGGACACCGAGCATTTGGCACGTCGCGCTCGATAA | 서열번호 2 |
명칭 | 서열 | 서열번호 |
GlxR_E45A | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGAPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 3 |
GlxR_E45S | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGSPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 4 |
GlxR_E45F | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGFPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 5 |
GlxR_E45K | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGKPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 6 |
GlxR_D66A | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGEPGDRLYIITSGKVKLARHAPAGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 7 |
GlxR_D66F | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGEPGDRLYIITSGKVKLARHAPFGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 8 |
GlxR_D66K | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGEPGDRLYIITSGKVKLARHAPKGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 9 |
GlxR_T166D | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGEPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGDQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 10 |
GlxR_T166F | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGEPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGFQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 11 |
GlxR_T166K | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGEPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGKQEAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 12 |
GlxR_E168A | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGEPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQAAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 13 |
GlxR_E168F | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGEPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQFAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 14 |
GlxR_E168K | MEGVQEILSRAGIFQGVDPTAVNNLIQDMETVRFPRGATIFDEGEPGDRLYIITSGKVKLARHAPDGRENLLTIMGPSDMFGELSIFDPGPRTSSAVCVTEVHAATMNSDMLRNWVADHPAIAEQLLRVLARRLRRTNASLADLIFTDVPGRVAKTLLQLANRFGTQKAGALRVNHDLTQEEIAQLVGASRETVNKALATFAHRGWIRLEGKSVLIVDTEHLARRAR | 서열번호 15 |
명칭 | 서열 (5’->3’) | 서열번호 |
glxR L1 | GAATTCGAGCTCGGTACCCTGAGACCCCACGCGAG | 서열번호 16 |
glxR R2 | GACTCTAGAGGATCCCCATCGACGGTGTCACCCCAC | 서열번호 17 |
glxR(E45A) L2 | GTAAAGGCGGTCACCTGGCgCGCCCTCGTCGAAGATGG | 서열번호 18 |
glxR(E45A) R1 | CCATCTTCGACGAGGGCGcGCCAGGTGACCGCCTTTAC | 서열번호 19 |
pDZ_F | TATTACGCCAGCTGGCGAAAG | 서열번호 20 |
pDZ_R | TTCCGGCTCGTATGTTGTGTG | 서열번호 21 |
glxR(E45S) L2 | GTAAAGGCGGTCACCTGGagaGCCCTCGTCGAAGATGG | 서열번호 22 |
glxR(E45S) R1 | CCATCTTCGACGAGGGCtctCCAGGTGACCGCCTTTAC | 서열번호 23 |
glxR(E45F) L2 | GTAAAGGCGGTCACCTGGaaaGCCCTCGTCGAAGATGG | 서열번호 24 |
glxR(E45F) R1 | CCATCTTCGACGAGGGCtttCCAGGTGACCGCCTTTAC | 서열번호 25 |
glxR(E45K) L2 | GTAAAGGCGGTCACCTGGCTtGCCCTCGTCGAAGATGG | 서열번호 26 |
glxR(E45K) R1 | CCATCTTCGACGAGGGCaAGCCAGGTGACCGCCTTTAC | 서열번호 27 |
glxR(D66A) L2 | GGTTTTCGCGGCCtgCCGGTGCGTGGCGCGCAAG | 서열번호 28 |
glxR(D66A) R1 | GCGCCACGCACCGGcaGGCCGCGAAAACCTGCTG | 서열번호 29 |
glxR(D66F) L2 | CAGCAGGTTTTCGCGGCCaaaCGGTGCGTGGCGC | 서열번호 30 |
glxR(D66F) R1 | GCGCCACGCACCGtttGGCCGCGAAAACCTGCTG | 서열번호 31 |
glxR(D66K) L2 | GGTTTTCGCGGCCtttCCGGTGCGTGGCGCGCAAG | 서열번호 32 |
glxR(D66K) R1 | GCGCCACGCACCGaaaGGCCGCGAAAACCTGCTG | 서열번호 33 |
glxR(T166D) L2 | GCCAGCTTCTTGGtcGCCGAAGCGGTTAGCCAGC | 서열번호 34 |
glxR(T166D) R1 | AACCGCTTCGGCgaCCAAGAAGCTGGCGCGCTG | 서열번호 35 |
glxR(T166F) L2 | GCCAGCTTCTTGGaaGCCGAAGCGGTTAGCCAGC | 서열번호 36 |
glxR(T166F) R1 | AACCGCTTCGGCttCCAAGAAGCTGGCGCGCTG | 서열번호 37 |
glxR(T166K) L2 | GCCAGCTTCTTGtttGCCGAAGCGGTTAGCCAGC | 서열번호 38 |
glxR(T166K) R1 | AACCGCTTCGGCaaaCAAGAAGCTGGCGCGCTG | 서열번호 39 |
glxR(E168A) L2 | AGCGCGCCAGCagCTTGGGCGCCGAAGCGGTTAG | 서열번호 40 |
glxR(E168A) R1 | CTTCGGCGCCCAAGctGCTGGCGCGCTGCGCGTG | 서열번호 41 |
glxR(E168F) L2 | AGCGCGCCAGCaaaTTGGGCGCCGAAGCGGTTAG | 서열번호 42 |
glxR(E168F) R1 | CTTCGGCGCCCAAtttGCTGGCGCGCTGCGCGTG | 서열번호 43 |
glxR(E168K) L2 | AGCGCGCCAGCcttTTGGGCGCCGAAGCGGTTAG | 서열번호 44 |
glxR(E168K) R1 | CTTCGGCGCCCAAaagGCTGGCGCGCTGCGCGTG | 서열번호 45 |
배지 종류 | 성분 |
종 배지 (pH 7.0) | 포도당 20 g, 펩톤 10 g, 효모추출물 5 g, 요소 1.5 g, KH2PO4 4 g, K2HPO4 8g, MgSO4·7H2O 0.5 g, 바이오틴 100 ㎍, 티아민 HCl 1000 ㎍, 칼슘-판토텐산 2000 ㎍, 니코틴아미드 2000 ㎍ (증류수 1 리터 기준) |
L-쓰레오닌 생산배지 (pH 7.2) | 포도당 30g, KH2PO4 2g, Urea 3g, (NH4)2SO4 40g, Peptone 2.5g, CSL(Sigma) 5g(10 ml), MgSO4·7H2O 0.5g, Leucine 400mg, CaCO3 20g(증류수 1 리터 기준) |
균주 | L-Thr 농도 (g/L) | 당 소모속도 (g/hr) | L-LYS 농도 (g/L) |
ATCC13032 | 0 | 3.65 | 1.2 |
ATCC13032 △glxR::glxR* (E45A) | 0.22 | 4.22 | 0 |
ATCC13032 △glxR::glxR* (D66A) | 0.16 | 3.98 | 0.2 |
ATCC13032 △glxR::glxR* (T166D) | 0.1 | - | 1 |
ATCC13032 △glxR::glxR* (E168A) | 0.06 | - | 0.7 |
균주 | L-Thr 농도 (g/L) | 당 소모속도 (g/hr) | L-LYS 농도 (g/L) |
ATCC13032 | 0 | 3.47 | 0.4 |
ATCC13032 △glxR::glxR* (E45A) | 0.2 | 4.02 | 0.4 |
ATCC13032 △glxR::glxR* (E45S) | 0.1 | 3.50 | 0.3 |
ATCC13032 △glxR::glxR* (E45F) | 0.1 | - | 0.4 |
ATCC13032 △glxR::glxR* (E45K) | 0.07 | 3.66 | 0.3 |
ATCC13032 △glxR::glxR* (D66A) | 0.22 | 3.76 | 0.2 |
ATCC13032 △glxR::glxR* (D66F) | 0.11 | - | 0.1 |
ATCC13032 △glxR::glxR* (D66K) | 0.06 | - | 0.4 |
ATCC13032 △glxR::glxR* (T166D) | 0.18 | 3.86 | 0.4 |
ATCC13032 △glxR::glxR* (T166F) | 0.07 | - | 0.3 |
ATCC13032 △glxR::glxR* (T166K) | 0.04 | 3.52 | 0.3 |
ATCC13032 △glxR::glxR* (E168A) | 0.15 | 3.81 | 0.3 |
ATCC13032 △glxR::glxR* (E168F) | 0.1 | - | 0.2 |
ATCC13032 △glxR::glxR* (E168K) | 0.05 | - | 0.1 |
명칭 | 서열 | 서열번호 |
LysC_L377K | MALVVQKYGGSSLESAERIRNVAERIVATKKAGNDVVVVCSAMGDTTDELLELAAAVNPVPPAREMDMLLTAGERISNALVAMAIESLGAEAQSFTGSQAGVLTTERHGNARIVDVTPGRVREALDEGKICIVAGFQGVNKETRDVTTLGRGGSDTTAVALAAALNADVCEIYSDVDGVYTADPRIVPNAQKLEKLSFEEMLELAAVGSKILVLRSVEYARAFNVPLRVRSSYSNDPGTLIAGSMEDIPVEEAVLTGVATDKSEAKVTVLGISDKPGEAAKVFRALADAEINIDMVLQNVSSVEDGTTDITFTCPRSDGRRAMEILKKLQVQGNWTNVLYDDQVGKVSLVGAGMKSHPGVTAEFMEALRDVNVNIEKISTSEIRISVLIREDDLDAAARALHEQFQLGGEDEAVVYAGTGR | 서열번호 46 |
LysC L1 | TCGAGCTCGGTACCCGCTGCGCAGTGTTGAATAC | 서열번호 47 |
LysC R2 | CTCTAGAGGATCCCCGTTCACCTCAGAGACGATT | 서열번호 48 |
lysC(L377K) L2 | TGGAAATCTTTTCGATGTTCACGTTGACAT | 서열번호 49 |
lysC(L377K) R1 | ATGTCAACGTGAACATCGAAAAGATTTCC | 서열번호 50 |
명칭 | 서열 | 서열번호 |
Hom_R407H | MTSASAPSFNPGKGPGSAVGIALLGFGTVGTEVMRLMTEYGDELAHRIGGPLEVRGIAVSDISKPREGVAPELLTEDAFALIEREDVDIVVEVIGGIEYPREVVLAALKAGKSVVTANKALVAAHSAELADAAEAANVDLYFEAAVAGAIPVVGPLRRSLAGDQIQSVMGIVNGTTNFILDAMDSTGADYADSLAEATRLGYAEADPTADVEGHDAASKAAILASIAFHTRVTADDVYCEGISNISAADIEAAQQAGHTIKLLAICEKFTNKEGKSAISARVHPTLLPVSHPLASVNKSFNAIFVEAEAAGRLMFYGNGAGGAPTASAVLGDVVGAARNKVHGGRAPGESTYANLPIADFGETTTRYHLDMDVEDRVGVLAELASLFSEQGISLRTIRQEERDDDAHLIVVTHSALESDLSRTVELLKAKPVVKAINSVIRLERD | 서열번호 51 |
hom L1 | ATCCTCTAGAGTCGACCCAACTGCAGACGTCGAA | 서열번호 52 |
hom R2 | ATGCCTGCAGGTCGACATAGACAGATTTGTCCACG | 서열번호 53 |
hom(R407H) L2 | GTGACCACGATCAGATGTGCATCATCATCGCGCTC | 서열번호 54 |
hom(R407H) R1 | GCGATGATGATGCACATCTGATCGTGGTCACCCAC | 서열번호 55 |
배지 종류 | 성분 |
종배지 (pH 7.0) | 포도당 20 g, 펩톤 10 g, 효모추출물 5 g, 요소 1.5 g, KH2PO4 4 g, K2HPO4 8 g, MgSO4·7H2O 0.5 g, 바이오틴 100 ㎍, 티아민 HCl 1000 ㎍, 칼슘-판토텐산 2000 ㎍, 니코틴아마이드 2000 ㎍ (증류수 1 리터 기준) |
L-쓰레오닌 생산배지 (pH 7.0) | 포도당 100g, KH2PO4 2g, Urea 3g, (NH4)2SO4 25g, Peptone 2.5g, CSL(Sigma) 5g(10 ml), MgSO4·7H2O 0.5g, 바이오틴 100 ㎍, 티아민 HCl 1000 ㎍, 칼슘-판토텐산 2000 ㎍, 니코틴아마이드 3000 ㎍, CaCO3 30g (증류수 1 리터 기준) |
균주 | L-Thr 농도 (g/L) | 당 소모속도 (g/hr) | L-LYS 농도 (g/L) |
KCCM12120P | 7.62 | 4.04 | 0.36 |
CA09-2373 | 9.23 | 4.66 | 0.28 |
CA09-2374 | 9.67 | 4.21 | 0.31 |
CA09-2325 | 8.87 | 4.13 | 0.35 |
CA09-2324 | 8.90 | 4.22 | - |
Claims (8)
- 하기 (1) 내지 (4)로 이루어진 군에서 선택된 변이를 포함하는, GlxR(glyoxylate bypass regulator) 단백질 변이체:(1) 서열번호 1의 아미노산 서열에서 N-말단으로부터 45번째 아미노산 위치에 상응하는 잔기가 알라닌(alanine), 세린(serine), 페닐알라닌(phenylalanine), 또는 라이신(lysine)으로 치환;(2) 서열번호 1의 아미노산 서열에서 N-말단으로부터 66번째 아미노산 위치에 상응하는 잔기가 알라닌(alanine), 페닐알라닌(phenylalanine), 또는 라이신(lysine)으로 치환;(3) 서열번호 1의 아미노산 서열에서 N-말단으로부터 166번째 아미노산 위치에 상응하는 잔기가 아스파르트산(aspartic acid), 페닐알라닌(phenylalanine), 또는 라이신(lysine)으로 치환; 및(4) 서열번호 1의 아미노산 서열에서 N-말단으로부터 168번째 아미노산 위치에 상응하는 잔기가 알라닌(alanine), 페닐알라닌(phenylalanine), 또는 라이신(lysine)으로 치환.
- 제1항의 GlxR 단백질 변이체를 암호화하는, 폴리뉴클레오티드.
- 제1항의 GlxR 단백질 변이체 및 상기 GlxR 단백질 변이체를 암호화하는 폴리뉴클레오티드로 이루어지는 군에서 선택된 1종 이상을 포함하는, 미생물.
- 제3항에 있어서, 상기 미생물은 L-쓰레오닌 생산능을 갖는 것인, 미생물.
- 제3항에 있어서, 상기 미생물은 코리네박테리움 속(Corynebacterium sp.) 인, 미생물.
- 제5항에 있어서, 상기 코리네박테리움 속 미생물은 코리네박테리움 글루타미쿰(Corynebacterium glutamicum)인, 미생물.
- 제3항 내지 제6항 중 어느 한 항의 미생물을 배지에서 배양하는 단계를 포함하는, L-쓰레오닌의 생산 방법.
- 제7항에 있어서, 상기 방법은 배양된 배지 또는 미생물에서 L-쓰레오닌을 회수하는 단계를 추가로 포함하는, L-쓰레오닌의 생산 방법.
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MX2023008105A MX2023008105A (es) | 2021-01-11 | 2022-01-06 | Variante de proteina glxr o metodo de produccion de treonina usando la misma. |
EP22736842.0A EP4276107A2 (en) | 2021-01-11 | 2022-01-06 | Glxr protein variant or threonine production method using same |
JP2023541941A JP2024503049A (ja) | 2021-01-11 | 2022-01-06 | GlxR蛋白質変異体またはこれを利用したスレオニン生産方法 |
CN202280009585.6A CN116888141A (zh) | 2021-01-11 | 2022-01-06 | Glxr蛋白变体或使用其生产苏氨酸的方法 |
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20080025355A (ko) | 2006-09-15 | 2008-03-20 | 씨제이제일제당 (주) | L-라이신 생산능이 향상된 코리네박테리아 및 그를이용한 l-라이신 생산 방법 |
US20160369310A1 (en) | 2014-09-05 | 2016-12-22 | Cj Cheiljedang Corporation | Microorganism with improved l-threonine producing capability, and method for producing l-threonine by using the same |
KR101947959B1 (ko) | 2018-05-28 | 2019-02-13 | 씨제이제일제당 (주) | 변이형 호모세린 디하이드로게나제 및 이를 이용한 호모세린 또는 호모세린 유래 l-아미노산의 생산 방법 |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1362866A1 (en) * | 2002-05-16 | 2003-11-19 | Basf Aktiengesellschaft | Use of the repressor glxR for the synthesis of lysine in Corynebacterium glutamicum |
KR100858913B1 (ko) * | 2007-03-09 | 2008-09-17 | 한국과학기술원 | 부위특이적 변이에 의해 제조된 고수율의 l-쓰레오닌생산균주 및 이를 이용한 l-쓰레오닌의 제조방법 |
KR101270149B1 (ko) * | 2009-11-25 | 2013-06-04 | 한국과학기술원 | L이소루신 고생성능을 가지는 변이미생물 및 이의 제조방법 |
WO2020138815A1 (ko) * | 2018-12-26 | 2020-07-02 | 대상 주식회사 | L-아미노산을 생산하는 대장균 변이주 또는 코리네박테리움 글루타미컴 변이주 및 이를 이용한 l-아미노산의 생산 방법 |
KR102221040B1 (ko) * | 2019-05-09 | 2021-03-03 | 씨제이제일제당 주식회사 | L-아미노산을 생산하는 미생물 및 이를 이용한 l-아미노산을 생산하는 방법 |
-
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20080025355A (ko) | 2006-09-15 | 2008-03-20 | 씨제이제일제당 (주) | L-라이신 생산능이 향상된 코리네박테리아 및 그를이용한 l-라이신 생산 방법 |
US20160369310A1 (en) | 2014-09-05 | 2016-12-22 | Cj Cheiljedang Corporation | Microorganism with improved l-threonine producing capability, and method for producing l-threonine by using the same |
KR101947959B1 (ko) | 2018-05-28 | 2019-02-13 | 씨제이제일제당 (주) | 변이형 호모세린 디하이드로게나제 및 이를 이용한 호모세린 또는 호모세린 유래 l-아미노산의 생산 방법 |
Non-Patent Citations (16)
Title |
---|
"Atlas Of Protein Sequence And Structure, National Biomedical Research Foundation", 1979, pages: 353 - 358 |
"GenBank", Database accession no. WP_003855810.1 |
"Guide to Huge Computers", 1994, ACADEMIC PRESS |
BLASTPBLASTNFASTAATSCHUL, J MOLEC BIOL, vol. 215, 1990, pages 403 |
CARILLO, SIAM J APPLIED MATH, vol. 48, 1988, pages 1073 |
DEVEREUX, J. ET AL., NUCLEIC ACIDS RESEARCH, vol. 12, 1984, pages 387 |
EMBOSS, HE EUROPEAN MOLECULAR BIOLOGY OPEN SOFTWARE SUITE |
EMBOSS, THE EUROPEAN MOLECULAR BIOLOGY OPEN SOFTWARE SUITE |
F.M. AUSUBEL ET AL.: "Manual of Methods for General Bacteriology", vol. 50-51, 1981, AMERICAN SOCIETY FOR BACTERIOLOGY, pages: 7 - 8 |
GRIBSKOV ET AL., NUCL. ACIDS RES, vol. 14, 1986, pages 6745 |
J. J.SAMBROOK ET AL.: "Molecular Cloning, A Laboratory Manual", 1989, COLD SPRING HARBOR LABORATORY PRESS |
NEEDLEMAN ET AL., J MOL BIOL., vol. 48, 1970, pages 443 |
NEEDLEMANWUNSCH, J. MOL. BIOL., vol. 48, 1970, pages 443 - 453 |
PEARSON ET AL., PROC. NATL. ACAD. SCI. USA, vol. 85, 1988, pages 2444 |
RICE ET AL., TRENDS GENET, vol. 16, 2000, pages 276 - 277 |
SMITHWATERMAN, ADV. APPL. MATH, vol. 2, 1981, pages 482 |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116120409A (zh) * | 2022-11-03 | 2023-05-16 | 华南理工大学 | 一种imp生物传感器及其构建方法与应用 |
CN116120409B (zh) * | 2022-11-03 | 2023-09-26 | 华南理工大学 | 一种imp生物传感器及其构建方法与应用 |
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EP4276107A2 (en) | 2023-11-15 |
JP2024503049A (ja) | 2024-01-24 |
WO2022149865A3 (ko) | 2022-08-25 |
AR124580A1 (es) | 2023-04-12 |
AU2022206623B2 (en) | 2024-01-04 |
MX2023008105A (es) | 2023-07-17 |
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