Functional analysis of arginine decarboxylase gene speA of Bacteroides dorei by markerless gene deletion. Issue 4 (8th January 2018)

• Record Type: Journal Article
• Authors: Sakanaka, Mikiyasu; Sugiyama, Yuta; Nara, Misaki; Kitakata, Aya; Kurihara, Shin
• Is Part Of: FEMS microbiology letters. Volume 365:Issue 4(2018:Feb.)
• Publication Date: 2018-01-08
• Subjects: arginine decarboxylase -- Bacteroides dorei -- gene manipulation system -- gut microbe -- polyamine -- spermidine; Microbiology -- Periodicals; 579
• DOI: 10.1093/femsle/fny003
• Languages: English
• ISSNs: 0378-1097

Abstract

Abstract: Polyamine concentrations in the intestine are regulated by their biosynthesis by hundreds of gut microbial species and these polyamines are involved in host health and disease. However, polyamine biosynthesis has not been sufficiently analyzed in major members of the human gut microbiota, possibly owing to a lack of gene manipulation systems. In this study, we successfully performed markerless gene deletion in Bacteroides dorei, one of the major members of the human gut microbiota. The combination of a thymidine kinase gene ( tdk ) deletion mutant and a counter-selection marker tdk, which has been applied in other Bacteroides species, was used for the markerless gene deletion. Deletion of tdk in B. dorei caused 5-fluoro-2 ΄ -deoxyuridine resistance, suggesting the utility of B. dorei Δ tdk as the host for future markerless gene deletions. Compared to parental strains, an arginine decarboxylase gene ( speA ) deletion mutant generated in this system showed a severe growth defect and decreased concentration of spermidine in the cells and culture supernatant. Collectively, our results indicate the accessibility of gene deletion and the important role of speA in polyamine biosynthesis in B. dorei . Abstract : Arginine decarboxylase gene speA was shown to be important for the growth of Bacteroides dorei and contributes to spermidine levels in cells and culture supernatant by markerless gene deletion.