Cloning and Characterization of the Gene Cluster Encoding Arthrofactin Synthetase from Pseudomonas sp. MIS38

Abstract

Arthrofactin is a potent cyclic lipopeptide-type biosurfactant produced by Pseudomonas sp. MIS38. In this work, an arthrofactin synthetase gene cluster (arf) spanning 38.7 kb was cloned and characterized. Three genes termed arfA, arfB, and arfC encode ArfA, ArfB, and ArfC, containing two, four, and five functional modules, respectively. Each module bears condensation, adenylation, and thiolation domains, like other nonribosomal peptide synthetases. However, unlike most of them, none of the 11 modules possess the epimerization domain responsible for the conversion of amino acid residues from L to D form. Possible L- and D-Leu adenylation domains specifically recognized only L-Leu. Moreover, two thioesterase domains are tandemly located at the C-terminal end of ArfC. These results suggest that ArfA, ArfB, and ArfC assemble to form a unique structure. Gene disruption of arfB impaired arthrofactin production, reduced swarming activity, and enhanced biofilm formation.

Antibacterial Activity of Marine Bacterium Pseudomonas sp. Associated with Soft Coral Sinularia polydactyla against Streptococcus equi Subsp. zooepidemicus

“…zooepidemicus. The members of genus Pseudomonas are well known to produce various peptide natural products including antibacterial (Mossialos el al., 2002), biosurfactants (Morikawa el al., 1993;Amada el al., 2000;Roongsang et al, 2003) and antifungals (Ramette et al, 2001;Nielsen el al., 2002;Sorensen el al., 2002).…”

Functional Analysis of A Pyoverdine Synthetase fromPseudomonassp. MIS38

“…MIS38. 5) Cloning vectors pGEM-T Easy and pBlueScript SK þ were purchased from Promega (Madison, WI) and Stratagene (La Jolla, CA) respectively. A kanamycin resistant gene (kan) was prepared from pSMC32 (Km r ), a derivative of pSU36.…”

“…Wild type MIS38 and mutant MT21 were grown at 30 C in L-broth for 40 h. An arthrofactin deficient mutant, NC1, 5) was used as a negative control. Extraction and analysis of arthrofactin were performed as previously described.…”

“…6,7) Such is the case for arthrofactin synthetase (ArfA/B/C). 5) When the primary structures of the Cdomains of L-peptidyl donors and D-peptidyl donors were compared, it was found that they belonged to different lineage.…”

Microbiotechnology Based Surfactants and Their Applications

“…It was initially postulated to be produced by an Arthrobacter sp. (Morikawa et al, 1993) but later corrected to originate from a Pseudomonas strain (Roongsawang et al, 2003). Cyclic lipopeptides that have been reported within the class Actinobacteria are the six amino acid containing cystargamide produced by Kitasatospora cystarginea (Gill et al, 2014) (20 , Table 9), the thirteen amino acid containing daptomycin produced by Streptomyces roseosporus (Debono et al, 1987) (21 , Table 9) and the depsipeptide ramoplanin, containing 16 amino acids, and which is produced by an Actinoplanes sp.…”