KR930021776A - Fermentation Method of Gene Production Strains by Addition of Organic Nitrogen Sources - Google Patents

Fermentation Method of Gene Production Strains by Addition of Organic Nitrogen Sources Download PDF

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KR930021776A
KR930021776A KR1019920005447A KR920005447A KR930021776A KR 930021776 A KR930021776 A KR 930021776A KR 1019920005447 A KR1019920005447 A KR 1019920005447A KR 920005447 A KR920005447 A KR 920005447A KR 930021776 A KR930021776 A KR 930021776A
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fermentation
yeast extract
cell
concentration
medium
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KR1019920005447A
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KR950009833B1 (en
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박태현
윤성관
최형배
김홍기
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최근선
주식회사 럭 키
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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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • 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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts

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Abstract

본 발명은 유가식 회분 발효에 의한 대장균 배양에 의해 유전자 제조합 단백질을 생산함에 있어서, 유기질소원인 효모 추출물을 함유하는 배지를 지수적으로 공급함으로써 프로데아제의 발현을 억제하는 단위 세포당 발현의 저하를 방지함을 특징으로 하는 발효 방법에 관한 것이다.In the present invention, in producing E. coli cultures by E. coli cultivation by fed-batch batch fermentation, it is possible to exponentially supply the medium containing yeast extract, which is an organic nitrogen source, to suppress the expression of proteases. A fermentation method characterized by preventing degradation.

Description

유기 질소원 첨가를 통한 유전자 제조합 균주의 발효 방법Fermentation Method of Gene Production Strains by Addition of Organic Nitrogen Sources

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음Since this is an open matter, no full text was included.

제1도는 효모 추출물이 공급되지 않는 유가식 회분 발효에서의 세포 성장 곡선과 단위 세포당 소 성장 호르몬 발현율을 나타낸 그래프이다.1 is a graph showing the cell growth curve and the expression rate of bovine growth hormone per unit cell in fed-batch batch fermentation without yeast extract.

제2도는 회분식 발효에서 PMSF가 단위 세포당 소 성장 호르몬 발현율에 미치는 영향을 나타낸 그래프이다,2 is a graph showing the effect of PMSF on the expression rate of bovine growth hormone per unit cell in batch fermentation.

제3도는 효모 추출물이 일시에 공급되는 유가식 발효에서의 세포성장 곡선과 단위 세포당 소 성장 호르몬 발현율을 나타낸 그래프이다.3 is a graph showing the cell growth curve and the expression rate of bovine growth hormone per unit cell in fed-batch fermentation in which yeast extract is supplied at one time.

제4도는 효모 추출물이 점진적으로 공급되는 유가식 회분 발효에서의 세포 성장 곡선과 단위 세포당 소 성장 호르몬 발현율을 나타낸 그래프이다.4 is a graph showing the cell growth curve and the expression rate of bovine growth hormone per unit cell in fed-batch batch fermentation in which yeast extract is gradually supplied.

Claims (6)

유가식 회분 발효에 의한 대장균 배양에 의해 유전자 제조합 단백질을 생산함에 있어서, 유기질소원인 효모 추출물을 함유하는 배지를 지수적으로 공급함으로써 프로테아제의 발현을 억제하여 단위 세포당 발현을 저하를 방지함을 특징으로 하는 발효 방법.In producing E. coli cultures by E. coli cultivation by fed-batch batch fermentation, by exponentially supplying a medium containing yeast extract, which is an organic nitrogen source, the expression of protease is suppressed to prevent a decrease in the expression per unit cell. Fermentation method characterized by. 제1항에 있어서, 배지는 다음 식에 의해 공급됨을 특징으로 하는 발효 방법.The fermentation method according to claim 1, wherein the medium is supplied by the following formula. F는 배지 공급 속도(ℓ/시간), μ는 제어하고자 하는 세포 비성장 속도(1/시간), X0는 초기 세포농도(g/ℓ), Y는 세포 성장수율(g세포/g포도당), SF는 공급배지내의 포도당농도(g/ℓ), VO는 초기 배양 부피를 나타내고 t는 시간을 나타낸다.F is the medium feed rate (l / hour), μ is the cell specific growth rate (1 / hour) to be controlled, X 0 is the initial cell concentration (g / l), and Y is the cell growth yield (g cell / g glucose). Where S F is the glucose concentration in the feed medium (g / L), V O represents the initial culture volume and t represents time. 제1항에 있어서, 발효 초기에는 효모 추출물을 포함하지 않은 공급 배지를 첨가하다가, 단위 세포당 발현을 저하가 발생하는 세포농도에 이르렀을 때 효모 추출물을 포함하는 공급 배지로 교환함을 특징으로 하는 발효 밥법.The method of claim 1, wherein in the initial stage of fermentation, a feed medium containing no yeast extract is added, and the expression per unit cell is changed to a feed medium containing yeast extract when the cell concentration at which a decrease occurs is reached. Fermented rice method. 제1항에 있어서, 효모 추출물을 포함한 공급 배지중의 효모 추출물 농도가 동일 배지내의 포도당 농도의 15 내지 40%인 공급 배지를 사용함을 특징으로 하는 발효 방법.The fermentation method according to claim 1, wherein the yeast extract concentration in the feed medium including the yeast extract is used as a feed medium in which 15 to 40% of the glucose concentration in the same medium is used. 제1항에 있어서, 효모 추출물을 포함한 공급 배지 중의 황산마그네슘(MgSO47H2O)농도가 동일 배지내의 포도당 농도의 2 내지 6%인 공급 배지를 사용함을 특징으로 하는 발효 방법.The fermentation method according to claim 1, wherein the magnesium sulfate (MgSO 4 7H 2 O) concentration in the feed medium containing the yeast extract is used with a feed medium having 2 to 6% of the glucose concentration in the same medium. 제1항에 있어서, 발효 말기에 효모 추출물의 시간당 최대 첨가 g수가, 리터로 환산한 발효 초기 부피 값의 5 내지 10 배에 해당하는 값을 갖는 것을 특징으로 하는 발효 방법.`The fermentation method according to claim 1, wherein the maximum number of added yeasts per hour of the yeast extract at the end of fermentation has a value corresponding to 5 to 10 times the initial volume value of fermentation converted into liters. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019920005447A 1992-04-01 1992-04-01 Culture method for tromsformed microorganism KR950009833B1 (en)

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KR1019920005447A KR950009833B1 (en) 1992-04-01 1992-04-01 Culture method for tromsformed microorganism

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KR930021776A true KR930021776A (en) 1993-11-23
KR950009833B1 KR950009833B1 (en) 1995-08-29

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