A Versatile Transcription-Translation in One Approach for Activation of Cryptic Biosynthetic Gene Clusters

doi:10.1021/acschembio.0c00581

CORC > 深海科学与工程研究所 > 中国科学院深海科学与工程研究所 > 深海科学研究部 > 深海生物学研究室 > 深海微生物活性物质研究组

	A Versatile Transcription-Translation in One Approach for Activation of Cryptic Biosynthetic Gene Clusters
	Zhang, Qian 3,4; Ren, Jin-Wei 2; Wang, Weishan 3,4; Zhai, Ji'an 1,4; Yang, Jing 3; Liu, Ning 3; Huang, Ying 3,4; Chen, Yihua 3,4; Pan, Guohui 3,4; Fan, Keqiang 3
刊名	ACS CHEMICAL BIOLOGY
	2020-09-18
卷号	15 期号:9 页码:2551-2557
ISSN号	1554-8929
DOI	10.1021/acschembio.0c00581
英文摘要	The ever-growing drug resistance problem worldwide highlights the urgency to discover and develop new drugs. Microbial natural products are a prolific source of drugs. Genome sequencing has revealed a tremendous amount of uncharacterized natural product biosynthetic gene clusters (BGCs) encoded within microbial genomes, most of which are cryptic or express at very low levels under standard culture conditions. Therefore, developing effective strategies to awaken these cryptic BGCs is of great interest for natural product discovery. In this study, we designed and validated a Transcription Translation in One (TTO) approach for activation of cryptic BGCs. This approach aims to alter the metabolite profiles of target strains by directly overexpressing exogenous rpsL (encoding ribosomal protein S12) and rpoB (encoding RNA polymerase beta subunit) genes containing beneficial mutations for natural product production using a plug-and-play plasmid system. As a result, this approach bypasses the tedious screening work and overcomes the false positive problem in the traditional ribosome engineering approach. In this work, the TTO approach was successfully applied to activating cryptic BGCs in three Streptomyces strains, leading to the discovery of two aromatic polyketide antibiotics, piloquinone and homopiloquinone. We further identified a single BGC responsible for the biosynthesis of both piloquinone and homopiloquinone, which features an unusual starter unit incorporation step. This powerful strategy can be further exploited for BGC activation in strains even beyond streptomycetes, thus facilitating natural product discovery research in the future.
资助项目	International Partnership Program of Chinese Academy of Sciences[153211KYSB20170014] ; program of China Ocean Mineral Resources RD Association[DY13S-B2-02] ; Senior User Project of RV KEXUE from the Center for Ocean Mega -Science, Chinese Academy of Sciences[KEXUE2019GZOS]
WOS关键词	STREPTOMYCES-COELICOLOR ; ANTIBIOTIC PRODUCTION ; NATURAL-PRODUCTS ; RNA-POLYMERASE ; POLYKETIDE ; MUTATIONS ; RESISTANCE ; DISCOVERY ; BACTERIAL ; PILOQUINONE
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000574920900024
资助机构	International Partnership Program of Chinese Academy of Sciences ; program of China Ocean Mineral Resources RD Association ; Senior User Project of RV KEXUE from the Center for Ocean Mega -Science, Chinese Academy of Sciences
内容类型	期刊论文
源URL	[http://ir.idsse.ac.cn/handle/183446/7951]
专题	深海科学研究部_深海生物学研究室_深海微生物活性物质研究组
通讯作者	Pan, Guohui; Fan, Keqiang
作者单位	1.Chinese Acad Sci, Inst Deep Sea Sci & Engn, Sanya 572000, Peoples R China 2.Chinese Acad Sci, Inst Microbiol, State Key Lab Mycol, Beijing 100101, Peoples R China 3.Chinese Acad Sci, Inst Microbiol, State Key Lab Microbial Resources, Beijing 100101, Peoples R China 4.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Qian,Ren, Jin-Wei,Wang, Weishan,et al. A Versatile Transcription-Translation in One Approach for Activation of Cryptic Biosynthetic Gene Clusters[J]. ACS CHEMICAL BIOLOGY,2020,15(9):2551-2557.
APA	Zhang, Qian.,Ren, Jin-Wei.,Wang, Weishan.,Zhai, Ji'an.,Yang, Jing.,...&Fan, Keqiang.(2020).A Versatile Transcription-Translation in One Approach for Activation of Cryptic Biosynthetic Gene Clusters.ACS CHEMICAL BIOLOGY,15(9),2551-2557.
MLA	Zhang, Qian,et al."A Versatile Transcription-Translation in One Approach for Activation of Cryptic Biosynthetic Gene Clusters".ACS CHEMICAL BIOLOGY 15.9(2020):2551-2557.