Document Detail


Isolation and characterization of a ferulic acid esterase (Fae1A) from the rumen fungus Anaeromyces mucronatus.
MedLine Citation:
PMID:  21362116     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Aims:  A novel ferulic acid esterase gene from rumen fungus Anaeromyces mucronatus was cloned, heteroexpressed in Escherichia coli and characterized. Methods and Results:  A total of 30 clones exhibiting activity on α-naphthyl acetate were isolated from an Anaeromyces mucronatus YE505 cDNA library. Sequence analysis revealed that these clones represented two esterase coding sequences. The gene, fae1A, showed highest amino acid sequence identity to CE family 1 esterases from anaerobic microorganisms such as Orpinomyces sp., Ruminococcus albus and Clostridium thermocellum. The gene comprised 828 nucleotides encoding a polypeptide of 275 amino acids. The coding sequence was cloned into the pET30a expression vector and over expressed in Escherichia coli BL21 (DE3). Gene product Fae1A was found to exhibit activity against a number of substrates including naphthyl fatty acid esters, p-nitrophenyl fatty acid esters and hydroxylcinnamic acid esters. Conclusions:  Fae1A exhibited a lower K(m) and higher catalytic efficiency (k(cat) /K(m) ) on ferulic acid esters than on α-naphthyl acetate or p-nitrophenyl acetate, suggesting that it has a higher affinity for ethyl and methyl ferulate than for the acetyl esters. It releases ferulic acid and p-coumaric acid from barley straw. Activity of Fae1A was inhibited by the serine specific protease inhibitor, phenylmethylsulfonyl fluoride, indicating that a serine residue plays a role in its activity. Significance and Impact of the Study:  To our knowledge, this is the first report of characterization of carbohydrate esterase gene from the genus of Anaeromyces.
Authors:
Meng Qi; Pan Wang; L Brent Selinger; L Jay Yanke; Robert J Forster; Tim A McAllister
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-3-1
Journal Detail:
Title:  Journal of applied microbiology     Volume:  -     ISSN:  1365-2672     ISO Abbreviation:  -     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-3-2     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:
Copyright © 2011 The Society for Applied Microbiology.
Affiliation:
Agriculture and Agri-Food Canada, Lethbridge Research Centre, Lethbridge, Alberta, Canada T1J 4B1 Department of Biological Science, University of Lethbridge, 4401 University Drive, Lethbridge, Alberta, Canada T1K 3M4.
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