A novel recombinant ethyl ferulate esterase from Burkholderia multivorans

摘要: Aims:  Isolation and identification of bacterial isolates with specific ferulic acid (FA) esterase activity cloning a gene encoding activity. Methods Results:  A micro-organism ethyl ferulate hydrolysing (EFH) was isolated by culture enrichment techniques. Detailed molecular based on species-specific primers two conserved genes (16S rRNA recA) led to the isolate as Burkholderia multivorans UWC10. (designated estEFH5) an EFH enzyme cloned its nucleotide sequence determined. Translational analysis revealed that estEFH5 encoded polypeptide 326 amino acids estimated weight 34·83 kDa. The EstEFH5 primary structure showed typical serine hydrolase motif (G-H-S-L-G). over-expressed in Escherichia coli insoluble form. Following urea denaturation vitro refolding, purified using one-step His Select™ Nickel chromatographic column. Conclusion:  Purified preference for short-chain ρ-nitrophenyl esters (C2 C3) feature carboxylesterase. Furthermore, recombinant also retained against (EF). Significance Impact Study:  biocatalytic process production FA from EF model substrate demonstrated. This is first report describes expression genus Burkholderia.