Improvement of clavulanic acid production in Streptomyces clavuligerus by genetic manipulation of structural biosynthesis genes

被引：12

作者：

Jnawali, Hum Nath ^{[2
]}

Yoo, Jin Cheol ^{[1
]}

Sohng, Jae Kyung ^{[2
]}

机构：

[1] Chosun Univ, Coll Pharm, Dept Pharm, Kwangju 501759, South Korea

[2] SunMoon Univ, Dept Pharmaceut Engn, Asan 336708, Chungnam, South Korea

来源：

BIOTECHNOLOGY LETTERS | 2011年 / 33卷 / 06期

关键词：

Clavulanic acid; Integration; Replication; Streptomyces clavuligerus; SYNTHASE;

D O I：

10.1007/s10529-011-0561-4

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

To enhance clavulanic acid production, four structural clavulanic acid biosynthesis genes, carboxyethylarginine synthase (ceas2), beta-lactam synthetase (bls2), clavaminate synthase (cas2) and proclavaminate amidinohydrolase (pah2), were amplified from Streptomyces clavuligerus genomic DNA. They were cloned in the pSET152 integration and pIBR25 expression vectors containing the strong ermE* promoter to generate pHN18 and pHN19, respectively, and both plasmids were introduced into S. clavuligerus by protoplast transformation. Clavulanic acid production was increased by 8.7-fold (to similar to 310 mg/l) in integrative pHN18 transformants and by 5.1-fold in pHN19 transformants compared to controls. Transcriptional analyses showed that the expression levels of ceas2, bls2, cas2 and pah2 were markedly increased in both transformants as compared with wild-type. The elevation of the ceas2, bls2, cas2 and pah2 transcripts was consistent with the enhanced production of clavulanic acid.

引用

页码：1221 / 1226

页数：6

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