caa a22 4500 605505055 CHVBK 20210128100619.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-6212-7 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6212-7 Effects of exogenous nutrients on polyketide biosynthesis in Escherichia coli [Elektronische Daten] [Lei Sun, Jia Zeng, Shuwei Zhang, Tyler Gladwin, Jixun Zhan] 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 1-6 were produced. While addition of glucose yielded the production of 1-4, 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 2-6 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. Springer-Verlag Berlin Heidelberg, 2014 Natural product biosynthesis nationallicence Type III polyketide synthase nationallicence Heterologous host nationallicence Escherichia coli nationallicence Exogenous nutrients nationallicence Sun Lei Department of Biological Engineering, Utah State University, 4105 Old Main Hill, 84322-4105, Logan, UT, USA aut Zeng Jia Department of Biological Engineering, Utah State University, 4105 Old Main Hill, 84322-4105, Logan, UT, USA aut Zhang Shuwei Department of Biological Engineering, Utah State University, 4105 Old Main Hill, 84322-4105, Logan, UT, USA aut Gladwin Tyler Department of Biological Engineering, Utah State University, 4105 Old Main Hill, 84322-4105, Logan, UT, USA aut Zhan Jixun Department of Biological Engineering, Utah State University, 4105 Old Main Hill, 84322-4105, Logan, UT, USA aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 693-701 0175-7598 99:2<693 2015 99 253 https://doi.org/10.1007/s00253-014-6212-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6212-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sun Lei Department of Biological Engineering, Utah State University, 4105 Old Main Hill, 84322-4105, Logan, UT, USA aut NATIONALLICENCE 700 1- Zeng Jia Department of Biological Engineering, Utah State University, 4105 Old Main Hill, 84322-4105, Logan, UT, USA aut NATIONALLICENCE 700 1- Zhang Shuwei Department of Biological Engineering, Utah State University, 4105 Old Main Hill, 84322-4105, Logan, UT, USA aut NATIONALLICENCE 700 1- Gladwin Tyler Department of Biological Engineering, Utah State University, 4105 Old Main Hill, 84322-4105, Logan, UT, USA aut NATIONALLICENCE 700 1- Zhan Jixun Department of Biological Engineering, Utah State University, 4105 Old Main Hill, 84322-4105, Logan, UT, USA aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 693-701 0175-7598 99:2<693 2015 99 253