Isolation and characterization of lipopeptide antibiotics produced by Bacillus subtilis

• Published in: Letters in applied microbiology Vol. 47; no. 3; pp. 180 - 186
• Main Authors: Chen, H., Wang, L., Su, C.X., Gong, G.H., Wang, P., Yu, Z.L.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.09.2008; Blackwell Science
• Subjects: Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - isolation & purification; Anti-Bacterial Agents - pharmacology; antibiotic; Bacillus subtilis; Bacillus subtilis - metabolism; Biological and medical sciences; characterization; Chromatography, High Pressure Liquid; electrospray ionization mass spectrometry; Fundamental and applied biological sciences. Psychology; lipopeptide; Lipopeptides - chemistry; Lipopeptides - isolation & purification; Lipopeptides - pharmacology; Microbiology; Spectrometry, Mass, Electrospray Ionization; electrospray ionization mass spectrometry; Bacillus subtilis; Applied microbiology; Bacillaceae; Electrospray; Characterization; Antibiotic; Bacillales; Ionization; Bacteria; Isolation; Mass spectrometry; Lipopeptide
• ISSN: 0266-8254; 1472-765X
• DOI: 10.1111/j.1472-765X.2008.02412.x

Aims:  Antibiotics from Bacillus subtilis JA show strong pathogen inhibition ability, which has potential market application; yet, the composition of these antibiotics has not been elucidated. The aim of this paper is to isolate and identify these antibiotics. Methods and Results:  The antagonistic activity of JA was tested in vitro; it exhibited strong inhibition against some important phytopathogens and postharvest pathogens. Crude antibiotic production was extracted with methanol from the precipitate by adding 6 mol l−1 HCl to the bacillus‐free culture broth. The crude extract was run on Diamonsil C18 column (5 μm, 250 × 4·6 mm) in HPLC system to separate the antibiotics. Major antibiotics were classified into three lipopeptide families according to electrospray ionization–mass spectrometry analysis. Subsequently, the classification of antibiotics was confirmed with typical collision‐induced dissociation fragments. Conclusions:  Three kinds of antibiotics were isolated from B. subtilis JA and were identified to the lipopeptide families, surfactin, iturin and fengycin. These compounds could function as biocontrol agents against a large spectrum of pathogens. Significance and Impact of the Study:  This study provided a reliable and rapid method for isolation and structural characterization of lipopeptide antibiotics from B. subtilis.