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- Title
Bioresolution of racemic phenyl glycidyl ether by a putative recombinant epoxide hydrolase from Streptomyces griseus NBRC 13350.
- Authors
Saini, Priya; Kumar, Naveen; Wani, Shadil; Sharma, Shilpi; Chimni, Swapandeep; Sareen, Dipti
- 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.
- Subjects
PHENYLGLYCIDYL ether; EPOXIDE hydrolase; STREPTOMYCES griseus; NATIONAL Board for Respiratory Care (U.S.); GENETIC overexpression; NUCLEOTIDE sequencing
- Publication
World Journal of Microbiology & Biotechnology, 2017, Vol 33, Issue 5, p1
- ISSN
0959-3993
- Publication type
Article
- DOI
10.1007/s11274-017-2248-z