Rashamuse, KJBurton, SGCowan, DA2009-05-072009-05-072007-11Rashamuse, KJ, Burton, SG and Cowan, DA. 2007. Novel recombinant ethyl ferulate esterase from Burkholderia multivorans. Journal of Applied Microbiology, Vol. (2007), pp 1-401364-5072http://hdl.handle.net/10204/3358This is a Post-print version of the work. It is posted here by permission of Blackwell Publishing for your personal use. Not for redistributionIsolation 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 BurkholderiaenEthyl ferulateGene cloningBurkholderia multivoransFerulic acid esteraseEsterase purificationNovel recombinant ethyl ferulate esteraseMicrobiologyArticleRashamuse, K., Burton, S., & Cowan, D. (2007). Novel recombinant ethyl ferulate esterase from Burkholderia multivorans. http://hdl.handle.net/10204/3358Rashamuse, KJ, SG Burton, and DA Cowan "Novel recombinant ethyl ferulate esterase from Burkholderia multivorans." (2007) http://hdl.handle.net/10204/3358Rashamuse K, Burton S, Cowan D. Novel recombinant ethyl ferulate esterase from Burkholderia multivorans. 2007; http://hdl.handle.net/10204/3358.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 -