Genome-wide investigation of the genes involved in nicotine metabolism in Pseudomonas putida J5 by Tn5 transposon mutagenesis

doi:10.1007/s00253-015-6529-x

	Genome-wide investigation of the genes involved in nicotine metabolism in Pseudomonas putida J5 by Tn5 transposon mutagenesis
	Xia, Zhenyuan 1; Zhang, Wei 2; Lei, Liping 1; Liu, Xingzhong 3; Wei, Hai-Lei 3
刊名	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
	2015
卷号	99 期号:15 页码:6503-6514
关键词	Nicotine Pseudomonas putida Biodegradation Tn5
ISSN号	0175-7598
DOI	10.1007/s00253-015-6529-x
通讯作者	Xia, Zhenyuan
英文摘要	Pseudomonas putida J5 is an efficient nicotine-degrading bacterial strain isolated from the tobacco rhizosphere. We successfully performed a comprehensive whole-genome analysis of nicotine metabolism-associated genes by Tn5 transposon mutagenesis in P. putida J5. A total of 18 mutants with unique insertions screened from 16,324 Tn5-transformants failed to use nicotine as the sole carbon source. Flanking sequences of the Tn5 transposon were cloned with a shotgun method from all of the nicotine-growth-deficient mutants. The potentially essential products of mutated gene were classified as follows: oxidoreductases, protein and metal transport systems, proteases and peptidases, transcriptional and translational regulators, and unknown proteins. Bioinformatic analysis of the Tn5 insertion sites indicated that the nicotine metabolic genes were separated and widely distributed in the genome. One of the mutants, M2022, was a Tn5 insert into a gene encoding a homolog of 6-hydroxy-l-nicotine oxidase, the second enzyme of nicotine metabolism in Arthrobacter nicotinovorans. Genetic and biochemical analysis confirmed that three open reading frames (ORFs) from an approximately 13-kb fragment recovered from the mutant M2022 were responsible for the transformation of nicotine to 3-succinoyl-pyridine via pseudooxynicotine and 3-succinoyl semialdehyde-pyridine, the first three steps of nicotine degradation. Further research on these mutants and the Tn5-inserted genes will help us characterize nicotine metabolic processes in P. putida J5.
学科主题	Biotechnology & Applied Microbiology
语种	英语
出版者	SPRINGER
WOS记录号	WOS:000357649200028
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/19748]
专题	农业环境与可持续发展研究所
作者单位	1.Yunnan Acad Tobacco Agr Sci, Kunming 650021, Yunnan, Peoples R China 2.Chinese Acad Agr Sci, Inst Environm & Sustainable Dev Agr, Key Lab Agr Environm, Minist Agr, Beijing 100081, Peoples R China 3.Chinese Acad Sci, Inst Microbiol, State Key Lab Mycol, Beijing 100101, Peoples R China
推荐引用方式 GB/T 7714	Xia, Zhenyuan,Zhang, Wei,Lei, Liping,et al. Genome-wide investigation of the genes involved in nicotine metabolism in Pseudomonas putida J5 by Tn5 transposon mutagenesis[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2015,99(15):6503-6514.
APA	Xia, Zhenyuan,Zhang, Wei,Lei, Liping,Liu, Xingzhong,&Wei, Hai-Lei.(2015).Genome-wide investigation of the genes involved in nicotine metabolism in Pseudomonas putida J5 by Tn5 transposon mutagenesis.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,99(15),6503-6514.
MLA	Xia, Zhenyuan,et al."Genome-wide investigation of the genes involved in nicotine metabolism in Pseudomonas putida J5 by Tn5 transposon mutagenesis".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 99.15(2015):6503-6514.