Zhang Haoran, Fang Lei, Osburne Marcia S, Pfeifer Blaine A
Department of Chemical and Biological Engineering, The State University of New York at Buffalo, 904 Furnas Hall, Buffalo, NY, 14260, USA.
EarthGenes Pharmaceuticals, Lexington, MA, 02421, USA.
Methods Mol Biol. 2016;1401:121-34. doi: 10.1007/978-1-4939-3375-4_8.
Heterologous biosynthesis of natural products is meant to enable access to the vast array of valuable properties associated with these compounds. Often motivated by limitations inherent in native production hosts, the heterologous biosynthetic process begins with a candidate host regarded as technically advanced relative to original producing organisms. Given this requirement, E. coli has been a top choice for heterologous biosynthesis attempts as associated recombinant tools emerged and continue to develop. However, success requires overcoming challenges associated with natural product formation, including complex biosynthetic pathways and the need for metabolic support. These two challenges have been heavily featured in cellular engineering efforts completed to position E. coli as a viable surrogate host. This chapter outlines steps taken to engineer E. coli with an emphasis on genetic manipulations designed to support the heterologous production of polyketide, nonribosomal peptide, and similarly complex natural products.
天然产物的异源生物合成旨在获取与这些化合物相关的大量宝贵特性。由于天然生产宿主存在固有限制，异源生物合成过程通常始于一种在技术上被认为比原始生产生物体更先进的候选宿主。基于这一要求，随着相关重组工具的出现并不断发展，大肠杆菌一直是异源生物合成尝试的首选。然而，要取得成功，需要克服与天然产物形成相关的挑战，包括复杂的生物合成途径以及对代谢支持的需求。在为将大肠杆菌定位为可行替代宿主而完成的细胞工程努力中，这两个挑战一直是重点。本章概述了对大肠杆菌进行工程改造所采取的步骤，重点是旨在支持聚酮化合物、非核糖体肽及类似复杂天然产物异源生产的基因操作。
