title: Bioresolution of racemic phenyl glycidyl ether by a putative recombinant epoxide hydrolase from Streptomyces griseus NBRC 13350.
abstract: In order to produce enantiomerically pure epoxides for the synthesis of value-added chemicals, a novel putative epoxide hydrolase (EH) sgeh was cloned and overexpressed in pET28a/Escherichia coli BL21(DE3). The 1047 bp sgeh gene was mined from Streptomyces griseus NBRC 13350 genome sequence. The recombinant hexahistidyl-tagged SGEH was purified (16.6-fold) by immobilized metal-affinity chromatography, with 90% yield as a homodimer of 100 kDa. The recombinant E. coli whole cells overexpressing SGEH could kinetically resolve racemic phenyl glycidyl ether (PGE) into (R)-PGE with 98% ee, 40% yield, and enantiomeric ratio (E) of 20. This was achieved under the optimized reaction conditions i.e. cell/substrate ratio of 20:1 (w/w) at pH 7.5 and 20 °C in 10% (v/v) dimethylformamide (DMF) in a 10 h reaction. 99% enantiopure (R)-PGE was obtained when the reaction time was prolonged to 12 h with a yield of 34%. In conclusion, an economically viable and environment friendly green process for the production of enantiopure (R)-PGE was developed by using wet cells of E. coli expressing recombinant SGEH.
authors.affiliation: Department of Biochemistry, Panjab University, Sector 25, Chandigarh, 160 014, India.
authors.forename: Priya
authors.initials: P
authors.lastname: Saini
authors.affiliation: Department of Chemistry, Guru Nanak Dev University, Amritsar, Punjab, 143 005, India.
authors.forename: Naveen
authors.initials: N
authors.lastname: Kumar
authors.affiliation: Department of Biochemistry, Panjab University, Sector 25, Chandigarh, 160 014, India.
authors.forename: Shadil Ibrahim
authors.initials: SI
authors.lastname: Wani
authors.affiliation: Department of Biochemistry, Panjab University, Sector 25, Chandigarh, 160 014, India.
authors.forename: Shilpi
authors.initials: S
authors.lastname: Sharma
authors.affiliation: Department of Chemistry, Guru Nanak Dev University, Amritsar, Punjab, 143 005, India.
authors.forename: Swapandeep Singh
authors.initials: SS
authors.lastname: Chimni
authors.affiliation: Department of Biochemistry, Panjab University, Sector 25, Chandigarh, 160 014, India. diptsare@pu.ac.in.
authors.forename: Dipti
authors.initials: D
authors.lastname: Sareen
mesh_terms: D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004851:Epoxide Hydrolases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D007700:Kinetics; D010647:Phenyl Ethers / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013237:Stereoisomerism; D013305:Streptomyces griseus / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D010647:Phenyl Ethers; D011994:Recombinant Proteins; C016050:phenylglycidyl ether; D004851:Epoxide Hydrolases
keywords: Bioresolution; Epoxide hydrolase; Phenyl glycidyl ether; Recombinant protein; Streptomyces griseus NBRC 13350