KR960007784A - Method for producing human alpha interferon by fed batch batch fermentation of recombinant E. coli - Google Patents

Method for producing human alpha interferon by fed batch batch fermentation of recombinant E. coli Download PDF

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Publication number
KR960007784A
KR960007784A KR1019940019963A KR19940019963A KR960007784A KR 960007784 A KR960007784 A KR 960007784A KR 1019940019963 A KR1019940019963 A KR 1019940019963A KR 19940019963 A KR19940019963 A KR 19940019963A KR 960007784 A KR960007784 A KR 960007784A
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South Korea
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arabinose
alpha interferon
medium
human alpha
recombinant
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KR1019940019963A
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Korean (ko)
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KR0134856B1 (en
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임형권
정경환
문흥모
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허영섭
재단법인 목암생명공학연구소
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    • 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
    • C12P21/00—Preparation of peptides or proteins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/555—Interferons [IFN]
    • C07K14/57—IFN-gamma

Abstract

본 발명은 인간알파 인터페론의 대량생산을 위해 살모넬라(Salmonell) 균주의 L-아라비노스(arabinose)오페론을 이용한 유도헝 프로모터가 포함된 유전자 재조합 대장균을 그라이셀롤과 효모추출물을 기본성분으로 하는 배지에서 배양하다가 일정한 시간에 L-아라비노스를 첨가하여 발현 유도를 시작한 후 배양액 내의 용준 산소량을 고갈 성분의 영양지표로 하여 배지를 공급함으로써 세포의 성장과 함께 지속적으로 재조합 유전자의 발현을 유도하여 발현된 재조합 인간 알파 인터페론이 세포내에 봉입체로서 다량 축적되게 하는 유가식 회분 발효방법을 제공한다.The present invention is to culture the recombinant E. coli containing the induced H. promoter using L-arabinose operon of Salmonell strain for mass production of human alpha interferon in a medium based on glycerol and yeast extract. Then, after inducing expression by adding L-arabinose at a certain time, and supplying a medium with the amount of dissolved oxygen in the culture medium as a nutrient depletion component, the recombinant human was expressed by continuously inducing the expression of the recombinant gene with the growth of the cells. A fed-batch batch fermentation method is provided in which a large amount of alpha interferon is accumulated as an inclusion body in a cell.

Description

재조합 대장균의 유가식 회분발효에 의한 인간 알파 인터페론의 제조방법Method for producing human alpha interferon by fed batch batch fermentation of recombinant E. coli

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

Claims (5)

(i)L-아라비노스 오페론을 이용한 유도성 발현벡터를 채용하는 재조합 미생물을 탄소원으로 글라이세롤 을 사용하는 초기배지에 배양하다가 대수 증식기 초기에 L-아라비노스를 첨가하여 연속적으로 발현을 유도하는 공정; 및 (ii)배양액내의 용존산소량의 변화를 영양성분의 고갈지표로 하여 글라이세롤을 성장제한 기질로 사용하는 유가배지를 주입하는 공정을 포함하는 L-아라비노스 오페론을 이용한 유도성 발현벡터를 채용하는 재조합 미생물의 고농동 배양방법.(i) Incubating recombinant microorganisms employing an inducible expression vector using L-arabinose operon in an initial medium using glycerol as a carbon source, and then continuously inducing expression by adding L-arabinose at the beginning of the logarithmic growth phase. fair; And (ii) injecting an oily medium using glycerol as a growth inhibitory substrate, using the change of dissolved oxygen in the culture medium as an indicator of nutrient depletion, and employing an inducible expression vector using L-Arabinose operon. High concentration culture method of recombinant microorganisms. (i)L-아라비노스 오페론을 이용한 유도성 발현벡터를 채용하는 재조합 미생물을 탄소원으로 글라이세롤 을 사용하는 초기배지에 배양하다가 대수 증식기 초기에 L-아라비노스를 첨가하여 연속적으로 발현을 유도하는 공정; 및 (ii)배양액내의 용존산소량의 변화를 영양성분의 고갈지표로 하여 글라이세롤을 성장제한 기질로 사용하는 유가배지를 주입하는 공정; 및 (iii)전기 미생물로부터 인간 알파 인터페론 활성을 지닌 단백질을 발현시키는 고정을 포함하는 인간 알파 인터페론의 제조방법.(i) Incubating recombinant microorganisms employing an inducible expression vector using L-arabinose operon in an initial medium using glycerol as a carbon source, and then continuously inducing expression by adding L-arabinose at the beginning of the logarithmic growth phase. fair; And (ii) injecting a milk medium using glycerol as a growth inhibitory substrate using the change in dissolved oxygen in the culture medium as an indicator of depletion of nutrients. And (iii) a fixation that expresses a protein having human alpha interferon activity from the electrical microorganism. 제2항에 있어서 L-아라비노스 오페론을 이용한 유도성 발현벡터를 채용하는 재조합 미생물은 Escherichia coli △pMAIFN α(KCCM 10053)인 것을 특징으로 하는 인간 알파 인터페론의 제조방법.The method for producing human alpha interferon according to claim 2, wherein the recombinant microorganism employing the inducible expression vector using L-arabinose operon is Escherichia coli ΔpMAIFN α (KCCM 10053). 제2항에 있어서 L-아라비노스는 배약액 1L당 2 내지 5g을 첨가하는 것을 특징으로 하는 인간 알파 인터페론의 제조방법.The method for preparing human alpha interferon according to claim 2, wherein L-arabinose is added in an amount of 2 to 5 g per 1 L of the aliquot. 제2항에 있어서 용존산소의 양애 50%이상이면 유가배지를 주입하고 용존산소의 양이 50%이하이면 유가배지의 주입을 중단하는 것을 특징으로 하는 인간 알파 인터페론의 제조방법.The method of claim 2, wherein if the amount of dissolved oxygen is greater than or equal to 50%, the fermented milk medium is injected, and if the amount of dissolved oxygen is less than 50%, the production of human alpha interferon is stopped. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019940019963A 1994-08-12 1994-08-12 Preparation process of human alpha interferon by fed-batch cultivation of recombinant e. coli KR0134856B1 (en)

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KR1019940019963A KR0134856B1 (en) 1994-08-12 1994-08-12 Preparation process of human alpha interferon by fed-batch cultivation of recombinant e. coli

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KR960007784A true KR960007784A (en) 1996-03-22
KR0134856B1 KR0134856B1 (en) 1998-04-14

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