期刊
ACS SYNTHETIC BIOLOGY
卷 3, 期 7, 页码 432-438出版社
AMER CHEMICAL SOC
DOI: 10.1021/sb400082j
关键词
valinomycin biosynthesis; nonribosornal peptide synthetase; heterologous expression; cyclodepsipeptide antibiotic; secondary metabolite production; synthetic biology
资金
- Berlin International Graduate School of Natural Sciences and Engineering (BIG-NSE) of the Cluster of Excellence Unifying Concepts in Catalysis - Deutsche Forschungsgemeinschaft (DFG)
- HaVo-Stiftung
The structural complexity of nonribosomal peptides (NRPs) impeding economic chemical synthesis and poor cultivability of source organisms limits the development of bioprocesses for novel bioactive compounds. Since nonribosomal peptide synthetases (NRPSs) assemble NRPs from simple amino acid building blocks, heterologous expression of NRPSs in a robust and easy to manipulate expression host is an attractive strategy to make pharmaceutically relevant NRPs more accessible and is also a basis for engineering of these enzymes to generate novel synthetic bioactive compounds. Here we show a systematic approach for the heterologous expression of the 654 kDa heterodimeric valinomycin synthetase (VlmSyn) from Streptomyces tsusimaensis in a soluble and active form in Escherichia coli. VlmSyn activity and precursor requirements were determined in vitro and provided evidence for a previously proposed model of valinomycin biosynthesis. In vivo production of recombinant valinomycin, a macrolactone antibiotic with reported antifungal, antibacterial, and antiviral activities, was achieved using an engineered E. coli strain growing in inexpensive media and independent of the supplementation with precursors and further optimization of the cultivation conditions. Tailoring of VlmSyn in E. coli paves the way to the production of novel valinomycin analogues in the future.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据