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PMID: 15345449 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cloning and characterization of the bile salt hydrolase genes (bsh) from Bifidobacterium bifidum strains.

Applied and environmental microbiology ·Vol. 70 ·No. 9 ·2004-09-00 ·Pages 5603-12

Kim GB, Miyamoto CM, Meighen EA, Lee BH

Abstract

Biochemical characterization of the purified bile salt hydrolase (BSH) from Bifidobacterium bifidum ATCC 11863 revealed some distinct characteristics not observed in other species of Bifidobacterium. The bsh gene was cloned from B. bifidum, and the DNA flanking the bsh gene was sequenced. Comparison of the deduced amino acid sequence of the cloned gene with previously known sequences revealed high homology with BSH enzymes from several microorganisms and penicillin V amidase (PVA) of Bacillus sphaericus. The proposed active sites of PVA were highly conserved, including that of the Cys-1 residue. The importance of the SH group in the N-terminal cysteine was confirmed by substitution of Cys with chemically and structurally similar residues, Ser or Thr, both of which resulted in an inactive enzyme. The transcriptional start point of the bsh gene has been determined by primer extension analysis. Unlike Bifidobacterium longum bsh, B. bifidum bsh was transcribed as a monocistronic unit, which was confirmed by Northern blot analysis. PCR amplification with the type-specific primer set revealed the high level of sequence homology in their bsh genes within the species of B. bifidum.

MeSH Terms
Amidohydrolases/genetics Amino Acid Sequence Base Sequence Bifidobacterium/enzymology,genetics Cloning, Molecular Consensus Sequence DNA Primers DNA, Bacterial/genetics Molecular Sequence Data RNA, Bacterial/genetics Sequence Alignment Sequence Homology, Amino Acid
Chemicals
DNA Primers DNA, Bacterial RNA, Bacterial Amidohydrolases choloylglycine hydrolase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kim Geun-Bae
Department of Food Science and Agricultural Chemistry, McGill University, 21111 Lakeshore Rd., Ste-Anne-de-Bellevue, Quebec H9X 3V9, Canada.
Miyamoto Carol M
Meighen Edward A
Lee Byong H
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.70.9.5603-5612.2004
Published
2004-09-00
Pages
5603-12
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC520925
Subset
IM
Databases
GENBANK
AY506536, AY604516, AY604517
External Links
PubMed source DOI: 10.1128/AEM.70.9.5603-5612.2004 PMC Full Text View journal details
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