子囊霉素工程菌的构建及初步发酵工艺优化  

Construction of engineering strain of ascomycin and optimization of fermentation process

作　　者：王志娟 黄鹤[1,2] 田勋 马宁 胡海峰 Wang Zhi-juan;Huang He;Tian Xun;Ma Ning;Hu Hai-feng(Shanghai Institute of Pharmaceutical Industry,China State Institute of Pharmaceutical Industry,Shanghai 200040;Sinopharm Health Industry Institute Co.,Ltd.,Shanghai 200437)

机构地区：[1]中国医药工业研究总院上海医药工业研究院,上海200040 [2]国药集团健康产业研究院有限公司,上海200437

出　　处：《中国抗生素杂志》2021年第9期854-858,共5页Chinese Journal of Antibiotics

摘　　要：基于子囊霉素的生物合成途径和机制,根据生物合成基因簇序列设计引物,应用PCR扩增游动放线菌(Antinoplanes sp.)N902-109赖氨酸合成途径关键基因——天冬氨酸激酶/天冬氨酸半醛脱氢酶/磷酸烯醇式丙酮酸羧化酶(lysC/asd/ppc)基因,整合至子囊霉素产生菌株吸水链霉菌(Streptomyces hygroscopicus)中,成功构建了过表达lysC/asd/ppc基因的重组工程菌,命名为AS-07。将AS-07工程菌进行摇瓶发酵研究,结果表明,与野生菌相比,子囊霉素的产量提高了25%。通过初步优化摇瓶发酵工艺,AS-07工程菌与野生菌相比,子囊霉素的产量提高了85%。Based on the biosynthetic pathway and mechanism of ascomycin,primers were designed according to the sequence of the biosynthetic gene cluster,using PCR amplification of Antinoplanes(Antinoplanes sp.)N902-109 keygenes of lysine synthesis pathway-aspartate kinase/aspartate semialdehyde dehydrogenase/phosphoenolpyruvate carboxylase(lysC/asd/ppc)genes,integration into the ascomycin producing strain Streptomyces hygroscopicus,a recombinant engineered strain overexpressing the lysC/asd/ppc genes were successfully constructed and named AS-07.The AS-07 engineered bacteria was subjected to shake flask fermentation research,and the results showed that compared with wild bacteria,the production of ascomycin was increased by 25%.Through preliminary optimization of the shake flask fermentation process,the production of ascomycin in AS-07 engineering bacteria was increased by 85%compared with wild bacteria.

关 键 词：子囊霉素 天冬氨酸激酶/天冬氨酸半醛脱氢酶/磷酸烯醇式丙酮酸羧(lys/asd/ppc)基因 基因工程菌 发酵工艺优化 

分 类 号：R978.15[医药卫生—药品] TQ465[医药卫生—药学]