增加ccaR基因剂量提高棒状链霉菌克拉维酸产量的研究  

STUDY ON AMPLIFICATION OF CCAR IN STREPTOMYCES CLAVULIGERUS TO ENHANCE CLAVULANIC ACID YIELD

作　　者：白小佳[1] 左志晗[1] 王艳萍[1] 

机构地区：[1]天津市食品营养与安全重点实验室,天津科技大学食品工程与生物技术学院,天津300457

出　　处：《食品研究与开发》2008年第8期67-70,共4页Food Research and Development

基　　金：天津市科委自然基金项目(043802711)

摘　　要：CcaR蛋白对棒状链霉菌(S.clavuligerus)中克拉维酸的合成具有正调控作用,由ccaR基因编码。PCR获得包含ccaR完整基因及其上下游一定区域的扩增产物,构建重组质粒pSET152-ccaR并转化大肠杆菌ET12567/pUZ8002后,通过属间接合转移至棒状链霉菌B71-14中。pSET152含有attP位点,可以与链霉菌基因组中的attB位点特异性同源整合,将重组质粒pSET152-ccaR携带的ccaR插入到S.clavuligerusB71-14染色体中的attB位点,实现了在S.clavuligerusB71-14基因组中增加一个拷贝ccaR基因的目的,所得的突变株S.clavuligerusB71-14:ccaR产酸量可达821.92 mg/L,是出发菌株的1.54倍。CcaR protein positively regulates clavulanic acid production in S.clavuligeru. In this work, ccaR gene was amplified by PCR. Plasmid pSET152-ccaR was constructed and transformed firstly to E.coli ET12567/ pUZ8002, and then transformed to S.clavuligerusB71-14 by conjugation. The attP site in pSET152-ccaR could integrated at attB site of S.clavuligerus B71-14, thus an extra copy ofccaR was inserted into the chromosome of S.clavuligerusB71-14. The clavulanic acid yield of the resulting strain S.clavuligerus B71-14： ccaR was 821.92 mg/L, which was 1.54 times that of the original strain S.clavuligerus B71-14.

关 键 词：棒状链霉菌 克拉维酸 ccaR基因 

分 类 号：TQ464[化学工程—制药化工]