摘要
裸盖菇素是一种精神活性生物碱,产自多种不同属的毒蕈,具有抗抑郁和抗焦虑的活性,可作为极有潜力的精神疾病治疗药物。基因组学技术的发展使裸盖菇素的生物合成基因与合成路线得以解析验证,本文以酿酒酵母为底盘细胞,运用CRISPR-Cas9多位点基因编辑技术将裸盖菇素合成基因Tdc、PsiK、PsiH、PsiM整合入酿酒酵母基因组上,并且通过底盘细胞菌株筛选以及合成基因拷贝数优化的方式有效提高其生物合成产量。结果显示,通过优选CEN.PK113-5D酵母菌株作为底盘细胞,并增加Tdc以及PsiH基因拷贝数,裸盖菇素的摇瓶产量达到114mg/L,是出发菌株的7.67倍。
Psilocybin is a psychoactive alkaloid derived from a variety of different genus of toadstools and has great potential as a drug for the treatment of psychiatric disorders.The development of genomics technology has enabled the analysis and verification of psilocybin biosynthesis genes and pathway.In this paper,CRISPR-Cas9 multi-site gene editing technology was used for chassis cell construction,and the biosynthesis genes Tdc,PsiK,PsiH and PsiM were integrated into the genome of Saccharomyces cerevisiae.Additionally,we achieve improved product yield through the screening of chassis cell strains and the optimization of the copy number of biosynthetic genes.The results show that the recombinant yeast strain for psilocybin production was successfully constructed,and psilocybin titers was improved effectively by means of rational chassis cell CEN.PK 113-5D and optimization of gene Tdc and PsiH copy number.At present,the shake flask yield of psilocybin reached 114 mg/L,which was 7.67times that of the initial strain.
作者
张鑫龙
万秀坤
王馥丽
姚戈
蒋辉
ZHANG Xinlong;WANG Xiukun;WANG Fuli;YAO Ge;JIANG Hui(State Key Laboratory of NBC Protection for Civilian,Beijing,102205)
出处
《生命科学仪器》
2022年第2期36-43,共8页
Life Science Instruments