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Please use this identifier to cite or link to this item: http://hdl.handle.net/10204/3358

Title: Novel recombinant ethyl ferulate esterase from Burkholderia multivorans
Authors: Rashamuse, KJ
Burton, SG
Cowan, DA
Keywords: Ethyl ferulate
Gene cloning
Burkholderia multivorans
Ferulic acid esterase
Esterase purification
Novel recombinant ethyl ferulate esterase
Microbiology
Issue Date: Nov-2007
Publisher: Blackwell Publishing
Citation: Rashamuse, KJ, Burton, SG and Cowan, DA. 2007. Novel recombinant ethyl ferulate esterase from Burkholderia multivorans. Journal of Applied Microbiology, Vol. (2007), pp 1-40
Abstract: Isolation and identification of bacterial isolates with specific ferulic acid (FA) esterase activity and cloning of a gene encoding activity. A micro-organism with ethyl ferulate hydrolysing (EFH) activity was isolated by culture enrichment techniques. Detailed molecular identification based on species-specific primers and two conserved genes (16S rRNA and recA) led to the identification of the isolate as Burkholderia multivorans UWC10. A gene (designated estEFH5) encoding an EFH enzyme was cloned and its nucleotide sequence determined. Translational analysis revealed that estEFH5 encoded a polypeptide of 326 amino acids with an estimated molecular weight of 34.83 kDa. The EstEFH5 primary structure showed a typical serine hydrolase motif (G-H-S-L-G). The estEFH5 gene was over-expressed in Escherichia coli in an insoluble form. Following urea denaturation and in vitro refolding, the enzyme was purified using one-step His Selectâ„¢ Nickel chromatographic column. Purified EstEFH5 showed a preference for short-chain -nitrophenyl esters (C2 and C3) a typical feature for carboxylesterase. Furthermore, the recombinant enzyme also retained the activity against ethyl ferulate (EF). Significance and Impact of the Study: A biocatalytic process for the production of FA from EF as a model substrate was demonstrated. This is the first report that describes the cloning and expression of a gene encoding FA esterase activity from the genus Burkholderia
Description: This is a Post-print version of the work. It is posted here by permission of Blackwell Publishing for your personal use. Not for redistribution
URI: http://hdl.handle.net/10204/3358
ISSN: 1364-5072
Appears in Collections:Enzyme technologies
General science, engineering & technology

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