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Molecular Cloning, Characterisation and Comparison of Bile Salt Hydrolases from Lactobacillus johnsonii PF01.
MedLine Citation:
PMID:  23035872     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
AIMS: To clone, characterise, and compare the bile salt hydrolase (BSH) genes of Lactobacillus johnsonii PF01. METHODS AND RESULTS: The BSH genes were amplified by polymerase chain reaction (PCR) using specific oligonucleotide primers, and the products were inserted into the pET21b expression vector. Escherichia coli BLR (DE3) cells were transformed with pET21b vectors containing the BSH genes and induced using 0.1 mmol l(-1) isopropylthiolgalactopyranoside (IPTG). The overexpressed BSH enzymes were purified using a nickel-nitrilotriacetic acid (Ni(2+) -NTA) agarose column and their activities characterised. BSH A hydrolysed tauro-conjugated bile salts optimally at pH 5.0 and 55°C, whereas BSH C hydrolysed glyco-conjugated bile salts optimally at pH 5.0 and 70°C. The enzymes had no preferential activities toward a specific cholyl moiety. CONCLUSIONS: BSH enzymes vary in their substrate specificities and characteristics to broaden its activity. Despite the lack of conservation in their putative substrate-binding sites, these remain functional through motif conservation. SIGNIFICANCE AND IMPACT OF THE STUDY: This is to our knowledge the first report of isolation of BSH enzymes from a single strain, showing hydrolase activity toward either glyco-conjugated or tauro-conjugated bile salts. Future structural homology studies and site-directed mutagenesis of sites associated with substrate specificity may elucidate specificities of BSH enzymes. © 2012The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.
Authors:
Jong Pyo Chae; Valerie Diane Valeriano; Geun-Bae Kim; Dae-Kyung Kang
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-10-4
Journal Detail:
Title:  Journal of applied microbiology     Volume:  -     ISSN:  1365-2672     ISO Abbreviation:  J. Appl. Microbiol.     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-10-5     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9706280     Medline TA:  J Appl Microbiol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
© 2012The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.
Affiliation:
Department of Animal Resources Science, Dankook University, Cheonan, Republic of Korea.
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