产青蒿酸酿酒酵母工程菌的抗性筛选  

Screening of Saccharomyces cerevisiae 1211 for Artemisinic Acid Production by Antibiotic Resistance

作　　者：陈伟 于文文 陈亚军 滕云 杨胜利[2] CHEN Wei;YU Wenwen;CHEN Yajun;TENG Yun;YANG Shengli(Zhejiang Key Lab. of Antifungal Drugs, Zhejiang Hisun Pharmaceutical Co., Ltd.,Taizhou 318000;College of Pharmacy, Zhejiang University of Technology, Hangzhou 310000)

机构地区：[1]浙江海正药业股份有限公司,浙江省抗真菌药物重点实验室,浙江台州318000 [2]浙江工业大学药学院,浙江杭州310000

出　　处：《中国医药工业杂志》2019年第5期524-529,共6页Chinese Journal of Pharmaceuticals

基　　金：国家科技重大专项子课题:青蒿素高效人工合成工艺开发和相关药物产业化研究(2017ZX09101002-003-003);中国博士后科学基金面上资助一等资助项目(2018M630690);浙江省教育厅一般科研项目(工程硕士专项)(Y201840669)

摘　　要：本研究以产青蒿酸的酿酒酵母工程菌为出发菌株,通过3 种抗菌素(诺尔斯菌素、遗传霉素和潮霉素B)对出发菌株进行抗性筛选。研究表明,当诺尔斯菌素和潮霉素B 复合添加且添加浓度分别为150 和100 μg/ml 时,经抗性筛选获得的抗性菌株发酵产量高达(926.8±21.9)mg/L,与出发菌株的(785.3±18.1)mg/L 相比提高了18.0%。经多次传代验证,经抗性筛选获得的菌株遗传稳定性好,具备工业化生产价值。The antibiotic resistance screening method was applied to screen strain with high-production of artemisinic acid from the original strain of Saccharomyces cerevisiae 1211. The effects of three antibiotics (nourseothricin sulfate, geneticin and hygromycin B) and their combinations on the inhibition of S. cerevisiae 1211 and the yield of artemisinic acid were investigated to obtain the highest yielding strain. The results showed that the yield of artemisinic acid produced by the strain treated with the combination of nourseothricin sulfate (150 μg/ml) and hygromycin B (100 μg/ml) achieved (926.8±21.9)mg/L, which was increased by 18 percent compared with the original strain [(785.3± 18.1)mg/L]. Furthermore, the strain obtained by resistance screening had excellent genetic stability by repeating subculture. In conclusion, this study provided a fast and effective way to screen engineered strain S. cerevisiae 1211 and showed the potential application to production of artemisinic acid.

关 键 词：青蒿酸 酿酒酵母工程菌 抗性筛选 遗传稳定性 

分 类 号：TQ920[轻工技术与工程—发酵工程]