Document Detail


Expression in yeast of secreted lignin peroxidase with improved 2,4-dichlorophenol degradability by DNA shuffling.
MedLine Citation:
PMID:  18514942     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Lignin peroxidase (LiP) from Phanerochaete chrysosporium was shown to mineralize a variety of recalcitrant aromatic compounds and oxidize a number of polycyclic aromatic and phenolic compounds. The major problem of the wild type LiP is that it can be inactivated by excess H(2)O(2) and high concentrations of aromatic compounds. We applied a directed evolution technique coupled with a rapid colorimetric screening method to obtain mutant genes with improved H(2)O(2) stability and polychlorinated phenol degradability, and they were successfully expressed as the secretive LiPs in recombinant Saccharomyces cerevisiae. The resulting variants showed approximately 1.6-fold improved 2,4-dichlorophenol (2,4-DCP) degradation activity and stability against H(2)O(2) compared with the parent strain. The kinetic properties of the variants toward 2,4-DCP and H(2)O(2) were also increased compared with the wild type for all three mutants studied. Amino acid sequence analysis indicated that the greatest number of amino acid substitutions was located near the surface or Ca(2+) binding sites of the enzyme.
Authors:
Kang Ryu; Jung Hye Kang; Lishi Wang; E K Lee
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-04-18
Journal Detail:
Title:  Journal of biotechnology     Volume:  135     ISSN:  0168-1656     ISO Abbreviation:  J. Biotechnol.     Publication Date:  2008 Jun 
Date Detail:
Created Date:  2008-06-23     Completed Date:  2008-10-06     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8411927     Medline TA:  J Biotechnol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  241-6     Citation Subset:  IM    
Affiliation:
Catholic Research Institute of Medical Science, College of Medicine, The Catholic University of Korea, 505 Banpo-dong, Seocho-gu, Seoul, Korea.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Substitution
Biodegradation, Environmental
Chlorophenols / metabolism*
Colorimetry
Culture Media
DNA Shuffling*
Kinetics
Mutation / genetics
Peroxidases / metabolism*,  secretion*
Phanerochaete / enzymology*
Protein Structure, Secondary
Saccharomyces cerevisiae / genetics*
Chemical
Reg. No./Substance:
0/Chlorophenols; 0/Culture Media; 120-83-2/2,4-dichlorophenol; EC 1.11.1.-/Peroxidases; EC 1.11.1.-/lignin peroxidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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