Measurement of bile salt hydrolase activity from Lactobacillus acidophilus based on disappearance of conjugated bile salts.
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 degrees 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.
Corzo
G
G
Department of Animal Science, Oklahoma State University, Stillwater 74078, USA.
Gilliland
S E
SE
D000581:Amidohydrolases / Q000032:analysis*; D001647:Bile Acids and Salts / Q000378:metabolism*; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid; D004492:Edetic Acid; D004795:Enzyme Stability; D005615:Freezing; D006000:Glycocholic Acid / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007779:Lactobacillus acidophilus / Q000201:enzymology*; D013656:Taurocholic Acid / Q000378:metabolism
D001647:Bile Acids and Salts; D013656:Taurocholic Acid; D004492:Edetic Acid; D000581:Amidohydrolases; C023154:choloylglycine hydrolase; D006000:Glycocholic Acid