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Purification and Characterization of an Intracellular β-Glucosidase from Lactobacillus casei ATCC 393

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Abstract

The lactic acid bacterium,Lactobacillus casei, produces an intracellular β-glucosidase when grown on Man-Rogosa-Sharpe (MRS) medium with cellobiose as carbon source. The β-glucosidase activity is produced intracellulary, and no extracellulary activity was detected. The enzyme was purified by ion-exchange chromatography and gel filtration. The molecular mass of the purified intracellular β-glucosidase as estimated by gel filtration was 480 kDa, consisting of six probably identical subunits. The enzyme exhibited optimum activity at 35°C and pH 6.3 with citrate-phosphate buffer. The enzyme was active against soluble glycosides with (1→4)-β configuration and from Lineweaver Burk plots, Km value of 16 mmol/L was found for β-pNPG. The β-glucosidase was competitively inhibited by glucose, and no glycosyl transferase activity was observed in the presence of ethanol.

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Authors and Affiliations

  1. Ecole Nationale Supérieure Agronomique de Montpellier, 2 place Pierre Viola, 34060 Montpellier Cedex 01, France

    S. Coulon, P. Chemardin, A. Arnaud & P. Galzy

  2. IFREMER Centre de Brest, Laboratoire de Biotechnologie, BP 70, 29280, Plouzane

    Y. Gueguen

Authors
  1. S. Coulon
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  2. P. Chemardin
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  3. Y. Gueguen
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  4. A. Arnaud
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  5. P. Galzy
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Coulon, S., Chemardin, P., Gueguen, Y. et al. Purification and Characterization of an Intracellular β-Glucosidase from Lactobacillus casei ATCC 393. Appl Biochem Biotechnol 74, 105–114 (1998). https://doi.org/10.1007/BF02787177

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  • DOI: https://doi.org/10.1007/BF02787177

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