Home | My Profile | Contact Us
Research Trends Products  |   order gateway  |   author gateway  |   editor gateway  
Register | Forgot Password

Author Resources
 Author Gateway
 Article submission guidelines

Editor Resources
 Editor/Referee Gateway

 Regional Subscription Agents/Distributors
Current Trends in Microbiology   Volumes    Volume 15 
Functional and biochemical analysis of recombinant N170V and R223F mutant bile salt hydrolases (BSHs) from Lactobacillus plantarum B14
Mehmet Öztürk, Zekiye Kılıçsaymaz
Pages: 55 - 62
Number of pages: 8
Current Trends in Microbiology
Volume 15 

Copyright © 2021 Research Trends. All rights reserved

Bile salt hydrolase (BSH), synthesized by some intestinal bacteria, catalyzes the hydrolysis of taurine and glycine-linked bile salts in human intestine. Because of the link between BSH and human health, the structural and biochemical features of BSHs necessitate an intensive study to better understand their roles in regulating bacterial and host metabolism. Five amino acids, supposed to be responsible for catalytic activity, are strictly conserved in all members of the N-terminal nucleophile (Ntn) hydrolase superfamily. In order to analyze the correlation between two of these strictly conserved amino acids and catalytic activity of BSH, the asparagine-170 (N170) and arginine-223 (R223) amino acids were substituted for the valine-170 (V170) and phenilalanine-223 (F223), respectively by polymerase chain reaction (PCR)-based site-directed mutagenesis. The mutant recombinant BSHs (mrBSHs) were expressed in Escherichia (E.) coli BLR(DE3). While the effect of the mutations on catalytic activity was detected by the ninhydrin assay, the effect of the mutations on formation of the BSHs was observed by sodium dodecyl sulfate poly acrylamide gel electrophoresis (SDS-PAGE) analysis. It was found that V170 and F223 mutations resulted in lack of BSH expression or stability. This is the first experimental report showing that the N170 and R223 may be critical amino acids for folding or stability of the BSH.
Buy this Article


Buy this article
Buy this volume
Subscribe to this title
Shopping Cart

Quick Links
Search Products
Browse in Alphabetical Order : Journals
Browse by Subject Classification : Journals

Ordering Information Ordering Information
Downloadable forms Downloadable Forms