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Bacillus spp. Produce Antibacterial Activities Against Lactic Acid Bacteria that Contaminate Fuel Ethanol Plants

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Abstract

Lactic acid bacteria (LAB) frequently contaminate commercial fuel ethanol fermentations, reducing yields and decreasing profitability of biofuel production. Microorganisms from environmental sources in different geographic regions of Thailand were tested for antibacterial activity against LAB. Four bacterial strains, designated as ALT3A, ALT3B, ALT17, and MR1, produced inhibitory effects on growth of LAB. Sequencing of rRNA identified these strains as species of Bacillus subtilis (ALT3A and ALT3B) and B. cereus (ALT17 and MR1). Cell mass from colonies and agar samples from inhibition zones were analyzed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The spectra of ALT3A and ALT3B showed a strong signal at m/z 1,060, similar in mass to the surfactin family of antimicrobial lipopeptides. ALT3A and ALT3B were analyzed by zymogram analysis using SDS-PAGE gels placed on agar plates inoculated with LAB. Cell lysates possessed an inhibitory protein of less than 10 kDa, consistent with the production of an antibacterial lipopeptide. Mass spectra of ALT17 and MR1 had notable signals at m/z 908 and 930 in the whole cell extracts and at m/z 687 in agar, but these masses do not correlate with those of previously reported antibacterial lipopeptides, and no antibacterial activity was detected by zymogram. The antibacterial activities produced by these strains may have application in the fuel ethanol industry as an alternative to antibiotics for prevention and control of bacterial contamination.

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Authors and Affiliations

  1. Biochemistry Unit, Department of Medical Sciences, Faculty of Science, Rangsit University, Muangake, Lakhok, Patumthani, 12000, Thailand

    Pennapa Manitchotpisit

  2. Renewable Product Technology Research Unit, National Center for Agricultural Utilization Research, Agricultural Research Service, U.S. Department of Agriculture, 1815 N. University St., Peoria, IL, 61604, USA

    Kenneth M. Bischoff, Neil P. J. Price & Timothy D. Leathers

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  1. Pennapa Manitchotpisit
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  2. Kenneth M. Bischoff
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  3. Neil P. J. Price
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  4. Timothy D. Leathers
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Correspondence to Kenneth M. Bischoff.

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Mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the United States Department of Agriculture and does not imply its approval to the exclusion of other products that may be suitable. USDA is an equal opportunity provider and employer.

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Manitchotpisit, P., Bischoff, K.M., Price, N.P.J. et al. Bacillus spp. Produce Antibacterial Activities Against Lactic Acid Bacteria that Contaminate Fuel Ethanol Plants. Curr Microbiol 66, 443–449 (2013). https://doi.org/10.1007/s00284-012-0291-4

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  • DOI: https://doi.org/10.1007/s00284-012-0291-4

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