Document Detail


Cloning, expression and decoding of the cold-adaptation of a new widely represented thermolabile subtilisin-like protease.
MedLine Citation:
PMID:  23043619     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
AIMS: Cloning, expression and characterization of a new cold-adapted protease with potential biotechnological application, isolated from Antarctic bacteria. METHOD AND RESULTS: A subtilisin-like gene was isolated from several Antarctic bacterial genus using CODPEHOP-designed primers and a genome walking method. This gene encodes a precursor protein which undergoes an autocatalytic cleavage resulting in a 34.6 kDa active cold-adapted protease with a maximum activity at 25-35ºC and optimum pH of 8.0-9.0. It showed a higher catalytic efficiency at lower temperatures compared to its mesophilic counterpart. Heat-induced inactivation resulted in a very low melting point. Local packing analysis using the homology model indicated Ala284 as an important cold-adaptation determinant, which was corroborated by the site-directed mutagenesis. CONCLUSIONS: A new thermolabile subtilisin-like protease has been successfully cloned and analyzed, and an important hotspot in the evolution of the cold-adaptation and substrate specificity of this enzyme was identified and tested. SIGNIFICANCE AND IMPACT OF THE STUDY: This work reports a new cold-adapted protease with a vast representation amongst Antarctic genus, suggesting therefore its evolutionary success in this cold environment. Likewise important sites for genetic potentiation have been identified, which are extrapolated to other enzymes of the same kind. © 2012The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.
Authors:
J P Acevedo; V Rodriguez; M Saavedra; M Muñoz; O Salazar; J A Asenjo; B A Andrews
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-10-9
Journal Detail:
Title:  Journal of applied microbiology     Volume:  -     ISSN:  1365-2672     ISO Abbreviation:  J. Appl. Microbiol.     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-10-9     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:
Facultad de Medicina y Facultad de Ingeniería y Ciencias Aplicadas, Universidad de los Andes, San Carlos de Apoquindo 2200, Santiago, Chile.
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