State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Chaoyang District, Beijing, 100101, People's Republic of China.
Shengxue Dacheng Pharmaceutical Co., Ltd., Shijiazhuang, 051430, Hebei, People's Republic of China.
Appl Microbiol Biotechnol. 2016 Dec;100(24):10563-10572. doi: 10.1007/s00253-016-7873-1. Epub 2016 Oct 5.
Heterologous expression is an important strategy to activate biosynthetic gene clusters of secondary metabolites. Here, it is employed to activate and manipulate the oxytetracycline (OTC) gene cluster and to alter OTC fermentation process. To achieve these goals, a fast-growing heterologous host Streptomyces venezuelae WVR2006 was rationally selected among several potential hosts. It shows rapid and dispersed growth and intrinsic high resistance to OTC. By manipulating the expression of two cluster-situated regulators (CSR) OtcR and OtrR and precursor supply, the OTC production level was significantly increased in this heterologous host from 75 to 431 mg/l only in 48 h, a level comparable to the native producer Streptomyces rimosus M4018 in 8 days. This work shows that S. venezuelae WVR2006 is a promising chassis for the production of secondary metabolites, and the engineered heterologous OTC producer has the potential to completely alter the fermentation process of OTC production.
异源表达是激活次生代谢物生物合成基因簇的重要策略。在这里，我们采用该策略来激活和操纵土霉素（OTC）基因簇，并改变 OTC 的发酵过程。为了实现这些目标，在几个潜在宿主中，我们合理地选择了生长迅速的异源宿主委内瑞拉链霉菌 WVR2006。该宿主表现出快速分散的生长和对 OTC 的固有高抗性。通过操纵两个簇定位调节因子（CSR）OtcR 和 OtrR 的表达和前体供应，仅在 48 小时内，OTC 的产量水平就从 75 毫克/升显著提高到 431 毫克/升，这一水平与土霉素的天然产生菌里氏木霉 M4018 在 8 天内的产量相当。这项工作表明，委内瑞拉链霉菌 WVR2006 是一种有前途的次生代谢产物生产底盘，工程化的异源 OTC 生产菌有可能彻底改变 OTC 生产的发酵过程。
