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Flexible exportation mechanisms of arthrofactin in Pseudomonas sp. MIS38

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Title: Flexible exportation mechanisms of arthrofactin in Pseudomonas sp. MIS38
Authors: Lim, S. P. Browse this author
Roongsawang, N. Browse this author
Washio, K. Browse this author →KAKEN DB
Morikawa, M. Browse this author →KAKEN DB
Keywords: ABC transporter
arthrofactin
biosurfactant
inhibitor
nonribosomal peptide
Pseudomonas
Issue Date: Jul-2009
Publisher: Blackwell Publishing
Journal Title: Journal of Applied Microbiology
Volume: 107
Issue: 1
Start Page: 157
End Page: 166
Publisher DOI: 10.1111/j.1365-2672.2009.04189.x
PMID: 19302333
Abstract: Aims: To obtain further insights into transportation mechanisms of a most effective biosurfactant, arthrofactin in Pseudomonas sp. MIS38. Methods and Results: A cluster genes arfA/B/C encodes an arthrofactin synthetase complex (ArfA/B/C). Downstream of the arfA/B/C lie genes encoding a putative periplasmic protein (ArfD, 362 aa) and a putative ATP-binding cassette transporter (ArfE, 651 aa), namely arfD and arfE, respectively. The arfA/B/C, arfD, and arfE form an operon suggesting their functional connection. Gene knockout mutants ArfD:Km, ArfE:Km, ArfD:Tc/ArfE:Km, and gene overexpression strains MIS38(pME6032_arfD/E) and ArfE:Km(pME6032_arfD/E) were prepared and analyzed for arthrofactin production profiles. It was found that the production levels of arthrofactin were temporally reduced in the mutants or increased in the gene overexpression strains, but they eventually became similar level to that of MIS38. Addition of ABC transporter inhibitors, glibenclamide and sodium ortho-vanadate dramatically reduced the production levels of arthrofactin. This excludes a possibility that arthrofactin is exported by diffusion with the aid of its own high surfactant activity. Conclusions: ArfD/E is not an exclusive but a primary exporter of arthrofactin during early growth stage. Reduction in the arthrofactin productivity of arfD and arfE knockout mutants was eventually rescued by another ABC transporter system. Effects of arfD and arfE overexpression were evident only for one-day cultivation. Multiple ATP dependent active transporter systems are responsible for the production of arthrofactin. Significance and impact of the study: Pseudomonas bacteria are characterized to be endued with multiple exporter and efflux systems for secondary metabolites including antibiotics, plant toxins, and biosurfactants. The present work demonstrates exceptionally flexible and highly controlled transportation mechanisms of a most effective lipopeptide biosurfactant, arthrofactin in Pseudomonas sp. MIS38. Because lipopeptide biosurfactants are known to enhance efficacy of bioactive compounds and arfA/B/C/D/E orthologous genes are also found in plant pathogenic P. fluorescens and P. syringae strains, the knowledge would also contribute to develop a technology controlling plant diseases.
Rights: The definitive version is available at www.blackwell-synergy.com
Type: article (author version)
URI: http://hdl.handle.net/2115/43176
Appears in Collections:環境科学院・地球環境科学研究院 (Graduate School of Environmental Science / Faculty of Environmental Earth Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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