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Measurement of Bile Salt Hydrolase Activity from Lactobacillus acidophilus Based on Disappearance of Conjugated Bile Salts1

  • Author Footnotes
    2 Present address: Suntory Institute of Biorganic Research (SUNBOR), Osaka, Japan.
    G. Corzo
    Footnotes
    2 Present address: Suntory Institute of Biorganic Research (SUNBOR), Osaka, Japan.
    Affiliations
    Department of Animal Science, Oklahoma State University, Stillwater 74078
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  • S.E. Gilliland
    Correspondence
    Corresponding author.
    Affiliations
    Department of Animal Science, Oklahoma State University, Stillwater 74078
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  • Author Footnotes
    1 Approved for publication by the director, Oklahoma Agricultural Experiment Station. This research was supported under Project H-2293. The senior author was supported on a CONACyTFulbright Fellowship.
    2 Present address: Suntory Institute of Biorganic Research (SUNBOR), Osaka, Japan.
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      Abstract

      Bile salt hydrolase activity of Lactobacillus acidophilus was measured based on the disappearance of sodium glycocholate and sodium taurocholate from the reaction mixture using HPLC. The amount of sodium glycocholate and sodium taurocholate that disappeared was proportional to the amount of sodium cholate that appeared in the mixture as detected by HPLC. Sodium glycocholate did not precipitate at the enzyme reaction conditions (37°C and pH 5.4) for determining bile salt hydrolase activity. The bile salt hydrolase assay was insensitive to low oxidation-reduction potential when measuring bile salt hydrolase from L. acidophilus, an intestinal microorganism. However, EDTA and freezing temperatures were necessary to maintain stability of the partially purified enzyme during storage.

      Key words

      Abbreviation key:

      BSH (bile salt hydrolase)

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