RU2015151124A - METHOD FOR PRODUCING L-AMINO ACIDS - Google Patents

METHOD FOR PRODUCING L-AMINO ACIDS Download PDF

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RU2015151124A
RU2015151124A RU2015151124A RU2015151124A RU2015151124A RU 2015151124 A RU2015151124 A RU 2015151124A RU 2015151124 A RU2015151124 A RU 2015151124A RU 2015151124 A RU2015151124 A RU 2015151124A RU 2015151124 A RU2015151124 A RU 2015151124A
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amino acids
producing
target gene
acids according
gene
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RU2015151124A
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Russian (ru)
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RU2614258C2 (en
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Дзун Ок МООН
Санг Дзо ЛИМ
До Хиун КВОН
Кванг Хо ЛИ
Хиун Вон БАЕ
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СиДжей ЧЕИЛДЗЕДАНГ КОРП.
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Priority claimed from PCT/KR2014/009468 external-priority patent/WO2015053552A1/en
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    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • 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
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/88—Lyases (4.)
    • 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
    • C12Y—ENZYMES
    • C12Y403/00—Carbon-nitrogen lyases (4.3)
    • C12Y403/03—Amine-lyases (4.3.3)
    • C12Y403/03007—4-Hydroxy-tetrahydrodipicolinate synthase (4.3.3.7)

Abstract

1. Способ получения L-аминокислот, включающий1) культивирование рекомбинантного коринеформного микроорганизма, способного продуцировать L-аминокислоты, где рекомбинантный коринеформный микроорганизм трансформируют посредством инсерции ацетат-индуцибельного промотора ниже стоп-кодона гена-мишени на хромосоме; и2) добавление ацетата при культивировании для ослабления экспрессии гена-мишени и усиления способности рекомбинантного коринеформного микроорганизма продуцировать L-аминокислоты,где область ниже стоп-кодона является областью, способной ослаблять экспрессию гена-мишени посредством приведения в действие ацетат-индуцируемого промотора в направлении, противоположном направлению транскрипции гена-мишени.2. Способ получения L-аминокислот по п. 1, где область ниже стоп-кодона находится между стоп-кодоном и областью выше терминатора транскрипции гена-мишени.3. Способ получения L-аминокислот по п. 1, где ацетат-индуцибельный промотор представлен нуклеотидной последовательностью SEQ ID NO: 1 или SEQ ID NO: 2.4. Способ получения L-аминокислот по п. 1, где ген-мишень является по меньшей мере одним геном, выбранным из группы, состоящей из гена, кодирующего субъединицу Е1 пируватдегидрогеназы, гена, кодирующего гомосериндегидрогеназу и гена, кодирующего УДФ-N-ацетилмурамоилаланил-D-глутамат-2,6-диаминопимелатлигазу.5. Способ получения L-аминокислот по п. 4, где L-аминокислота является L-лизином.6. Способ получения L-аминокислот по п. 1, где ген-мишень является геном, кодирующим дигидродипиколинатсинтазу.7. Способ получения L-аминокислот по п. 6, где L-аминокислота является L-треонином.1. A method for producing L-amino acids, including 1) culturing a recombinant coryneform microorganism capable of producing L-amino acids, where the recombinant coryneform microorganism is transformed by insertion of an acetate-inducible promoter below the stop codon of the target gene on the chromosome; and 2) adding acetate during cultivation to attenuate the expression of the target gene and enhance the ability of the recombinant coryneform microorganism to produce L-amino acids, where the region below the stop codon is the region capable of attenuating the expression of the target gene by driving the acetate-induced promoter in the opposite direction the direction of transcription of the target gene. 2. A method for producing L-amino acids according to claim 1, wherein the region below the stop codon is between the stop codon and the region above the transcription terminator of the target gene. The method for producing L-amino acids according to claim 1, wherein the acetate-inducible promoter is represented by the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2.4. The method for producing L-amino acids according to claim 1, wherein the target gene is at least one gene selected from the group consisting of a gene encoding the E1 subunit of pyruvate dehydrogenase, a gene encoding a homoserine dehydrogenase and a gene encoding UDF-N-acetylmuramoylalanyl-D- glutamate-2,6-diaminopimelatligase. 5. A method for producing L-amino acids according to claim 4, wherein the L-amino acid is L-lysine. A method for producing L-amino acids according to claim 1, wherein the target gene is a gene encoding a dihydrodipicolinate synthase. The method for producing L-amino acids according to claim 6, wherein the L-amino acid is L-threonine.

Claims (7)

1. Способ получения L-аминокислот, включающий1. The method of obtaining L-amino acids, including 1) культивирование рекомбинантного коринеформного микроорганизма, способного продуцировать L-аминокислоты, где рекомбинантный коринеформный микроорганизм трансформируют посредством инсерции ацетат-индуцибельного промотора ниже стоп-кодона гена-мишени на хромосоме; и1) the cultivation of a recombinant coryneform microorganism capable of producing L-amino acids, where the recombinant coryneform microorganism is transformed by insertion of an acetate-inducible promoter below the stop codon of the target gene on the chromosome; and 2) добавление ацетата при культивировании для ослабления экспрессии гена-мишени и усиления способности рекомбинантного коринеформного микроорганизма продуцировать L-аминокислоты,2) the addition of acetate during cultivation to weaken the expression of the target gene and enhance the ability of the recombinant coryneform microorganism to produce L-amino acids, где область ниже стоп-кодона является областью, способной ослаблять экспрессию гена-мишени посредством приведения в действие ацетат-индуцируемого промотора в направлении, противоположном направлению транскрипции гена-мишени.where the region below the stop codon is a region capable of attenuating the expression of the target gene by activating the acetate-induced promoter in the direction opposite to the direction of transcription of the target gene. 2. Способ получения L-аминокислот по п. 1, где область ниже стоп-кодона находится между стоп-кодоном и областью выше терминатора транскрипции гена-мишени.2. A method for producing L-amino acids according to claim 1, wherein the region below the stop codon is between the stop codon and the region above the transcription terminator of the target gene. 3. Способ получения L-аминокислот по п. 1, где ацетат-индуцибельный промотор представлен нуклеотидной последовательностью SEQ ID NO: 1 или SEQ ID NO: 2.3. The method for producing L-amino acids according to claim 1, wherein the acetate-inducible promoter is represented by the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2. 4. Способ получения L-аминокислот по п. 1, где ген-мишень является по меньшей мере одним геном, выбранным из группы, состоящей из гена, кодирующего субъединицу Е1 пируватдегидрогеназы, гена, кодирующего гомосериндегидрогеназу и гена, кодирующего УДФ-N-ацетилмурамоилаланил-D-глутамат-2,6-диаминопимелатлигазу.4. The method of producing L-amino acids according to claim 1, where the target gene is at least one gene selected from the group consisting of a gene encoding the E1 subunit of pyruvate dehydrogenase, a gene encoding a homoserine dehydrogenase and a gene encoding UDF-N-acetylmuramoylalanyl- D-glutamate-2,6-diaminopimelatligase. 5. Способ получения L-аминокислот по п. 4, где L-аминокислота является L-лизином.5. The method for producing L-amino acids according to claim 4, wherein the L-amino acid is L-lysine. 6. Способ получения L-аминокислот по п. 1, где ген-мишень является геном, кодирующим дигидродипиколинатсинтазу.6. The method for producing L-amino acids according to claim 1, wherein the target gene is a gene encoding a dihydrodipicolinate synthase. 7. Способ получения L-аминокислот по п. 6, где L-аминокислота является L-треонином. 7. The method for producing L-amino acids according to claim 6, wherein the L-amino acid is L-threonine.
RU2015151124A 2013-10-11 2014-10-08 Method of producing l-amino acids RU2614258C2 (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
KR20130121090 2013-10-11
KR10-2013-0121090 2013-10-11
KR1020140091307A KR101518860B1 (en) 2013-10-11 2014-07-18 Method for producing L-amino acid
KR10-2014-0091307 2014-07-18
PCT/KR2014/009468 WO2015053552A1 (en) 2013-10-11 2014-10-08 Method of producing l-amino acids

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KR101518860B1 (en) * 2013-10-11 2015-05-12 씨제이제일제당 (주) Method for producing L-amino acid
CN105734004B (en) * 2016-03-02 2020-06-19 廊坊梅花生物技术开发有限公司 Recombinant strain and preparation method and application thereof
KR101925930B1 (en) * 2017-07-24 2019-02-26 씨제이제일제당 주식회사 Microorganisms producing L-lysine and process for producing L-lysine using the same
WO2019172702A1 (en) * 2018-03-09 2019-09-12 씨제이제일제당 (주) Novel promoter and l-amino acid production method using same
KR102221040B1 (en) * 2019-05-09 2021-03-03 씨제이제일제당 주식회사 Microorganism producing L-amino acid and method of producing Method of L-amino acid using thereof
KR102207867B1 (en) * 2020-01-21 2021-01-26 씨제이제일제당 주식회사 Method for producing L-amino acids using microorganisms containing NADP dependent glyceraldehyde-3-phosphate dehydrogenase

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PH12016500640A1 (en) 2016-05-30
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JO3255B1 (en) 2018-09-16
RU2016110804A (en) 2016-07-27
BR112016008029A2 (en) 2017-10-03
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RU2614258C2 (en) 2017-03-24
JP2018113966A (en) 2018-07-26
CA2925659C (en) 2018-10-23
JP6824918B2 (en) 2021-02-03
UA117935C2 (en) 2018-10-25
AR097977A1 (en) 2016-04-20
JP6291042B2 (en) 2018-03-14
ES2626048T3 (en) 2017-07-21
DK2860256T3 (en) 2017-06-06
CN104561163A (en) 2015-04-29
AU2017272295B2 (en) 2019-05-16
AU2014332732A1 (en) 2016-04-21
JP2020078312A (en) 2020-05-28
PH12016500640B1 (en) 2016-05-30
KR101518860B1 (en) 2015-05-12
GEP20196943B (en) 2019-02-11
AU2017272295A1 (en) 2018-01-04
KR20150042692A (en) 2015-04-21
AU2014332732B2 (en) 2017-12-07
RU2651461C2 (en) 2018-04-19
CA2925659A1 (en) 2015-04-16

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