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Purification and Characterization of Three Different Types of Bile Salt Hydrolases from Bifidobacterium Strains

¹ Department of Food Science and Agricultural Chemistry, McGill University, 21,111 Lakeshore Road, Ste-Anne-de-Bellevue, QC, Canada H9X 3V9
² Food Research & Development Centre, Agriculture and Agri-Food Canada, St-Hyacinthe, QC, Canada J2S 8E3

Corresponding author: B. H. Lee; e-mail: blee{at}macdonald.mcgill.ca.

Bile salt hydrolases were purified to electrophoretic homogeneity from Bifidobacterium bifidum ATCC 11863, Bifidobacterium infantis KL412, Bifidobacterium longum ATCC 15708, Bifidobacterium longum KL507, and Bifidobacterium longum KL515. Three different types (A, B, and C) of bile salt hydrolase (BSH) were revealed during the purification study, exhibiting the type-specific characteristics in their electrophoretic migration and elution profiles from anion exchange and hydrophobic interaction chromatographic columns. The subunit molecular mass estimated by sodium dodecylsulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) was around 35 kDa, and the native molecular mass in all five Bifidobacterium strains was estimated to be between 130 and 150 kDa by gel filtration chromatography, indicating that all BSH enzymes have tetrameric structure. From the isoelectric focusing, an isoelectric point value of 4.45 was obtained with BSH (type B) from B. bifidum ATCC 11863 and the other BSH (types A and C) showed the similar pI values around 4.65. N-Terminal amino acid sequencing for the proteins of types A and C revealed that 6 out of 20 amino acid residues were different, and highly conserved residues were identified in both N-terminal sequences of types A and C. All BSH enzymes from five strains hydrolyzed six major human bile salts, and they showed a better deconjugation rate on glycine-conjugated bile salts than on taurine-conjugated forms.

Key Words: bile salt hydrolase • purification • Bifidobacterium

Abbreviation key: BSH = bile salt hydrolase, FPLC = fast protein liquid chromatography, HIC = hydrophobic interaction chromatography, IEC = ion-exchange chromatography, IEF = isoelectric focusing, LAB = lactic acid bacteria, pI = isoelectric point, SEC = size exclusion chromatography

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