Recombineering for Genetic Engineering of Natural Product Biosynthetic Pathways

Muhammad Nazeer Abbasi; Jun Fu; Xiaoying Bian; Hailong Wang; Youming Zhang; Aiying Li

doi:10.1016/j.tibtech.2019.12.018

Recombineering for Genetic Engineering of Natural Product Biosynthetic Pathways

Trends Biotechnol. 2020 Jul;38(7):715-728. doi: 10.1016/j.tibtech.2019.12.018. Epub 2020 Jan 20.

Authors

Muhammad Nazeer Abbasi¹, Jun Fu¹, Xiaoying Bian¹, Hailong Wang², Youming Zhang³, Aiying Li⁴

Affiliations

¹ Helmholtz International Lab for Anti-Infectives, Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, PR China.
² Helmholtz International Lab for Anti-Infectives, Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, PR China. Electronic address: wanghailong@sdu.edu.cn.
³ Helmholtz International Lab for Anti-Infectives, Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, PR China. Electronic address: zhangyouming@sdu.edu.cn.
⁴ Helmholtz International Lab for Anti-Infectives, Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, PR China. Electronic address: ayli@sdu.edu.cn.

PMID: 31973879
DOI: 10.1016/j.tibtech.2019.12.018

Abstract

Microbial genomes encode many cryptic and uncharacterized biosynthetic gene clusters (BGCs). Exploiting this unexplored genetic wealth to discover microbial novel natural products (NPs) remains a challenging issue. We review homologous recombination (HR)-based recombineering, mediated by the recombinases RecE/RecT from Rac prophage and Redα/Redβ from lambda phage, which has developed into a highly inclusive tool for direct cloning of large DNA up to 100 kb, seamless mutation, multifragment assembly, and heterologous expression of microbial NP BGCs. Its utilization in the refactoring, engineering, and functional expression of long BGCs for NP biosynthesis makes it easy to elucidate NP-producing potential in microbes. This review also highlights various applications of recombineering in NP-derived drug discovery.

Keywords: Recombineering; biosynthetic gene cluster; direct cloning; homologous recombination; natural product.

Publication types

Research Support, Non-U.S. Gov't
Review

MeSH terms

Biological Products / chemistry
Biological Products / metabolism*
Biosynthetic Pathways / genetics*
Genetic Engineering*
Homologous Recombination / genetics
Metabolic Engineering*
Multigene Family

Substances

Biological Products