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Characterization of Lactose Utilization and β-Galactosidase in Lactobacillus brevis KB290, the Hetero-Fermentative Lactic Acid Bacterium

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Abstract

Unlike dairy lactic acid bacteria, Lactobacillus brevis cannot ferment milk. We characterized the lactose utilization by L. brevis KB290. In a carbohydrate fermentation assay using API 50 CHL, we showed during 7 days L. brevis did not ferment lactose. L. brevis grew to the stationary phase in 2 weeks in MRS broth containing lactose as the carbon source. L. brevis slowly consumed the lactose in the medium. L. brevis hydrolyzed lactose and a lactose analog, o-nitrophenyl-β-d-galactopyranoside (ONPGal). This β-galactosidase activity for ONPGal was not repressed by glucose, galactose, fructose, xylose, or maltose showing the microorganism may not have carbon catabolite repression. We purified the L. brevis β-galactosidase using ammonium sulfate precipitation and several chromatographies. The enzyme’s molecular weight is estimated at 72 and 37 kDa using SDS-PAGE analysis. The N-terminal amino acid sequence of the larger protein was 90 % similar to the sequence of the putative β-galactosidase (YP_796339) and the smaller protein was identical to the sequence of the putative β-galactosidase (YP_796338) in L. brevis ATCC367. This suggests the enzyme is a heterodimeric β-galactosidase. The specific activity of the purified enzyme for lactose is 55 U/mg. We speculate inhibition of lactose transport delays the lactose utilization in L. brevis KB290.

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Acknowledgments

We would like to thank Dr. Yuki Sawatari and Mr. Naoto Takahashi for valuable discussions.

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Authors and Affiliations

  1. Research Institute, Kagome Co., Ltd., 17 Nishitomiyama, Nasushiobara, 329-2762, Japan

    Hiroyuki Honda & Nobuhiro Yajima

  2. Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba, Sendai, 981-8555, Japan

    Tadao Saito

Authors
  1. Hiroyuki Honda
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  2. Nobuhiro Yajima
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  3. Tadao Saito
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Correspondence to Hiroyuki Honda.

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Honda, H., Yajima, N. & Saito, T. Characterization of Lactose Utilization and β-Galactosidase in Lactobacillus brevis KB290, the Hetero-Fermentative Lactic Acid Bacterium. Curr Microbiol 65, 679–685 (2012). https://doi.org/10.1007/s00284-012-0216-2

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  • DOI: https://doi.org/10.1007/s00284-012-0216-2

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