Development and characterization of recombinant whole-cell biocatalysts expressing epoxide hydrolase from Rhodotorula glutinis for enantioselective resolution of racemic epoxides

Comparative homology modeling-inspired protein engineering for improvement of catalytic activity of Mugil cephalus epoxide hydrolase

“…Hence, we carried out homology modelinginspired protein engineering to enhance the activity of M. cephalus EH. Because the sequence similarity of M. cephalus EH with A. niger EH using PredictProtein server (http://www.predictprotein.org/) is about 30%, the active site of M. cephalus EH was successfully modeled on the basis of A. niger EH (Kim et al 2006;Nardini et al 1999;Zou et al 2000). The modeled 3-D structure of the active site of M. cephalus EH is shown in Fig.…”

“…The reaction time required for the completion of kinetic resolution was decreased up to 26-fold by homology modeling-inspired triple point mutations. Compared to microbial EH, the hydrolytic activity of triple mutants was all most similar to that of the recombinant E. coli cells expressing the microbial EH gene of Rhodotorula glutinis (Kim et al 2006). When the enantioselective hydrolysis reaction was carried out in the presence of phenyl-1,2-ethanediol up to The mutated amino acid residues and nucleophilic Asp residue are indicated by arrow.…”

Biocatalysis as an alternative for the production of chiral epoxides: A comparative review

“…However, genes encoding various types of EHs have been cloned and expressed in heterologous hosts and these recombinant EHs have also been applied, which opens up more opportunities to control the product quality and purity [152,175], as well as to manipulate the enzymes at genetic level to improve their properties [189][190][191]. A recent report on a recombinant EH from Rhodutorula glutinis expressed in S. cerevisiae and E. coli showed that the reaction system could tolerate a styrene oxide concentration of 40 mM, resulting in (S)-styrene oxide with an enantiomeric excess higher than 99% and 21.3% yield [153,154].…”

Enantioselective resolution of racemic styrene oxide at high concentration using recombinant Pichia pastoris expressing epoxide hydrolase of Rhodotorula glutinis in the presence of surfactant and glycerol

“…The recombinant P. pastoris was cultivated on a BMGY medium containing 10 g yeast extract/l and 10 g peptone/l at 30°C (Kim et al 2006). The expression of EH gene was induced by adding 1% (v/v) methanol for 72 h. The cells were harvested, and then washed with Tris/HCl buffer (100 mM, pH 8).…”