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Novel recombinant ethyl ferulate esterase from Burkholderia multivorans

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dc.contributor.author Rashamuse, KJ
dc.contributor.author Burton, SG
dc.contributor.author Cowan, DA
dc.date.accessioned 2009-05-07T14:20:19Z
dc.date.available 2009-05-07T14:20:19Z
dc.date.issued 2007-11
dc.identifier.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 en
dc.identifier.issn 1364-5072
dc.identifier.uri http://hdl.handle.net/10204/3358
dc.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 en
dc.description.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 Select™ 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 en
dc.language.iso en en
dc.publisher Blackwell Publishing en
dc.subject Ethyl ferulate en
dc.subject Gene cloning en
dc.subject Burkholderia multivorans en
dc.subject Ferulic acid esterase en
dc.subject Esterase purification en
dc.subject Novel recombinant ethyl ferulate esterase en
dc.subject Microbiology en
dc.title Novel recombinant ethyl ferulate esterase from Burkholderia multivorans en
dc.type Article en
dc.identifier.apacitation Rashamuse, K., Burton, S., & Cowan, D. (2007). Novel recombinant ethyl ferulate esterase from Burkholderia multivorans. http://hdl.handle.net/10204/3358 en_ZA
dc.identifier.chicagocitation Rashamuse, KJ, SG Burton, and DA Cowan "Novel recombinant ethyl ferulate esterase from Burkholderia multivorans." (2007) http://hdl.handle.net/10204/3358 en_ZA
dc.identifier.vancouvercitation Rashamuse K, Burton S, Cowan D. Novel recombinant ethyl ferulate esterase from Burkholderia multivorans. 2007; http://hdl.handle.net/10204/3358. en_ZA
dc.identifier.ris TY - Article AU - Rashamuse, KJ AU - Burton, SG AU - Cowan, DA AB - 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 Select™ 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 DA - 2007-11 DB - ResearchSpace DP - CSIR KW - Ethyl ferulate KW - Gene cloning KW - Burkholderia multivorans KW - Ferulic acid esterase KW - Esterase purification KW - Novel recombinant ethyl ferulate esterase KW - Microbiology LK - https://researchspace.csir.co.za PY - 2007 SM - 1364-5072 T1 - Novel recombinant ethyl ferulate esterase from Burkholderia multivorans TI - Novel recombinant ethyl ferulate esterase from Burkholderia multivorans UR - http://hdl.handle.net/10204/3358 ER - en_ZA


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