Effects of exogenous nutrients on polyketide biosynthesis in Escherichia coli

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• 作者：Lei Sun ; Jia Zeng ; Shuwei Zhang ; Tyler Gladwin…
• 关键词：Natural product biosynthesis ; Type III polyketide synthase ; Heterologous host ; Escherichia coli ; Exogenous nutrients
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2015
• 出版时间：January 2015
• 年：2015
• 卷：99
• 期：2
• 页码：693-701
• 全文大小：1,603 KB
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• 作者单位：Lei Sun (1)
  Jia Zeng (1)
  Shuwei Zhang (1)
  Tyler Gladwin (1)
  Jixun Zhan (1)
  
  1. Department of Biological Engineering, Utah State University, 4105 Old Main Hill, Logan, UT, 84322-4105, USA
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

Heterologous hosts are important platforms for engineering natural product biosynthesis. Escherichia coli is such a host widely used for expression of various biosynthetic enzymes. While numerous studies have been focused on optimizing the expression conditions for desired functional proteins, this work describes how supplement of exogenous nutrients into the fermentation broth influences the formation of natural products in E. coli. A type III polyketide synthase gene stts from Streptomyces toxytricini NRRL 15443 was heterogeneously expressed in E. coli BL21(DE3). This enzyme uses five units of malonyl-CoA to generate a polyketide 1,3,6,8-tetrahydroxynaphthalene, which can be spontaneously oxidized into a red compound flaviolin. In this work, we manipulated the fermentation broth of E. coli BL21(DE3)/pET28a-stts by supplying different nutrients including glucose and sodium pyruvate at different concentrations, from which six flaviolin derivatives 1-strong class="a-plus-plus">6 were produced. While addition of glucose yielded the production of 1-strong class="a-plus-plus">4, supplement of sodium pyruvate into the induced broth of E. coli BL21(DE3)/pET28a-stts resulted in the synthesis of 5 and 6, suggesting that different nutrients may enable E. coli to generate different metabolites. These products were purified and structurally characterized based on the spectral data, among which 2-strong class="a-plus-plus">6 are novel compounds. These molecules were formed through addition of different moieties such as acetone and indole to the flaviolin scaffold. The concentrations of glucose and sodium pyruvate and incubation time affect the product profiles. This work demonstrates that supplement of nutrients can link certain intracellular metabolites to the engineered biosynthetic pathway to yield new products. It provides a new approach to biosynthesizing novel molecules in the commonly used heterologous host E. coli.