Abstract
Lactic acid bacteria (LAB) contamination of beer presents a continual economic threat to brewers. Interestingly, only certain isolates of LAB can grow in the hostile beer environment (e.g., as studied here, Lactobacillus brevis BSO 464 (Lb464) and a non-ropy isolate of Pediococcus claussenii ATCC BAA-344T (Pc344NR)), indicating that significant genetic specialization is required. The genes hitA, horA, horB, horC, and bsrA, which have been proposed to confer beer-spoiling ability to an organism, are suspected of counteracting the antimicrobial effects of hops. However, these genes are not present in the same combination (if at all) across beer-spoiling organisms. As such, we sought to investigate the extent to which these genes participate during Lb464 and Pc344NR mid-logarithmic growth in beer through reverse transcription quantitative PCR analysis. We first determined the optimal reference gene set needed for data normalization and, for each bacterium, established that two genes were needed for accurate assessment of gene expression. Following this, we found that horA expression was induced for Pc344NR, but not for Lb464, during growth in beer. Instead, horC expression was dramatically increased in Lb464 when growing in beer, whereas no change was detected for the other putative beer-spoilage-related genes. This indicates that HorC may be one of the principle mediators enabling growth of Lb464 in beer, whereas in Pc344NR, this may be attributable to HorA. These findings not only reveal that Lb464 and Pc344NR are unique in their beer-specific genetic expression profile but also indicate that a range of genetic specialization exists among beer-spoilage bacteria.
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Acknowledgements
JB was the recipient of a College of Medicine, University of Saskatchewan Graduate Scholarship and a College of Graduate Studies and Research, University of Saskatchewan, Graduate Scholarship. VP was the holder of an Alexander Graham Bell Canada Graduate Scholarship and a Michael Smith Foreign Study Supplement from the Natural Sciences and Engineering Research Council of Canada (NSERC) and also the recipient of the MillerCoors, Anheuser-Busch InBev, and Brian Williams Graduate Scholarships from the American Society of Brewing Chemists Foundation. This research was supported by MillerCoors Brewing Company, Golden, Colorado, and NSERC Discovery Grant 24067.
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Jordyn Bergsveinson and Vanessa Pittet contributed equally.
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Bergsveinson, J., Pittet, V. & Ziola, B. RT-qPCR analysis of putative beer-spoilage gene expression during growth of Lactobacillus brevis BSO 464 and Pediococcus claussenii ATCC BAA-344T in beer. Appl Microbiol Biotechnol 96, 461–470 (2012). https://doi.org/10.1007/s00253-012-4334-3
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DOI: https://doi.org/10.1007/s00253-012-4334-3