Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.
Engineering Research Center of Bioconversion and Bio-purification, Ministry of Education, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.
Microb Cell Fact. 2020 Jan 31;19(1):18. doi: 10.1186/s12934-020-1290-y.
Amphotericin B (AmB) is widely used against fungal infection and produced mainly by Streptomyces nodosus. Various intracellular metabolites of S. nodosus were identified during AmB fermentation, and the key compounds that related to the cell growth and biosynthesis of AmB were analyzed by principal component analysis (PCA) and partial least squares (PLS).
Rational design that based on the results of metabolomics was employed to improve the AmB productivity of Streptomyces nodosus, including the overexpression of genes involved in oxygen-taking, precursor-acquiring and product-exporting. The AmB yield of modified strain S. nodosus VMR4A was 6.58 g/L, which was increased significantly in comparison with that of strain S. nodosus ZJB2016050 (5.16 g/L). This was the highest yield of AmB reported so far, and meanwhile, the amount of by-product amphotericin A (AmA) was decreased by 45%. Moreover, the fermentation time of strain S. nodosus VMR4A was shortened by 24 h compared with that of strain. The results indicated that strain S. nodosus VMR4A was an excellent candidate for the industrial production of AmB because of its high production yield, low by-product content and the fast cell growth.
This study would lay the foundation for improving the AmB productivity through metabolomics analysis and overexpression of key enzymes.
两性霉素 B（AmB）被广泛用于抗真菌感染，主要由游动放线菌产生。在 AmB 发酵过程中鉴定了游动放线菌的各种细胞内代谢物，并通过主成分分析（PCA）和偏最小二乘法（PLS）分析与 AmB 细胞生长和生物合成相关的关键化合物。
基于代谢组学的结果进行了合理的设计，以提高游动放线菌的 AmB 产量，包括参与吸氧、前体获取和产物输出的基因的过表达。改良菌株游动放线菌 VMR4A 的 AmB 产量为 6.58 g/L，与游动放线菌 ZJB2016050（5.16 g/L）相比，显著增加。这是迄今为止报道的最高 AmB 产量，同时，副产物两性霉素 A（AmA）的量减少了 45%。此外，与菌株相比，菌株游动放线菌 VMR4A 的发酵时间缩短了 24 h。结果表明，由于其高生产产量、低副产物含量和快速细胞生长，菌株游动放线菌 VMR4A 是 AmB 工业生产的优秀候选菌株。
本研究将为通过代谢组学分析和关键酶的过表达来提高 AmB 生产力奠定基础。
