Liu_2013_Appl.Microbiol.Biotechnol_97_7659

Reference

Title : Enzymatic production of Cilastatin intermediate via highly enantioselective hydrolysis of methyl (+\/-)-2,2-dimethylcyclopropane carboxylate using newly isolated Rhodococcus sp. ECU1013 - Liu_2013_Appl.Microbiol.Biotechnol_97_7659
Author(s) : Liu CH , Pan J , Ye Q , Xu JH
Ref : Applied Microbiology & Biotechnology , 97 :7659 , 2013
Abstract :

(S)-(+)-2,2-Dimethylcyclopropane carboxylic acid [(S)-(+)-DMCPA] is a key chiral intermediate for production of Cilastatin, an excellent renal dehydropeptidase-I inhibitor. In this study, a new method for preparation of (S)-(+)-DMCPA with microbial esterases was investigated. A microbial screening program obtained six esterase-producing isolates that could display relatively high activities and enantioselectivities using racemic ethyl 2,2-dimethylcyclopropane carboxylate (DMCPE) as screening substrate, aiming at forming optically pure (S)-(+)-DMCPA. Further selection was carried out with substrates having different alcohol moieties, including methyl, ethyl, and 2-chloroethyl esters. Finally, one of these strains, numbered ECU1013, with high enantioselectivity toward the hydrolytic resolution of methyl 2,2-dimethylcyclopropane carboxylate (DMCPM), afforded the (S)-product in 92 % ee, and was later identified as Rhodococcus sp. According to our research, there were several active esterases to DMCPM in cells of Rhodococcus sp. ECU1013; however, (S)-preferential esterase was selectively enriched based on the time-dependent profile of esterases biosynthesis, thereby the enantiomeric excess of biotransformation product (ee p) was constantly increased, finally maintained at 95 % (S). To improve the yield, various organic solvents were employed for better dispersion of the hydrophobic substrate. As a result, (+/-)-DMCPM of up to 400 mM in the organic phase of isooctane was enantioselectively hydrolyzed into (S)-(+)-DMCPA, with an isolation yield of 38 % and a further increase of ee p to 99 %.

PubMedSearch : Liu_2013_Appl.Microbiol.Biotechnol_97_7659
PubMedID: 23807665
Gene_locus related to this paper: rhosp-AHY95170

Related information

Substrate DMCPE
Gene_locus DMCPE    rhosp-AHY95170

Citations formats

Liu CH, Pan J, Ye Q, Xu JH (2013)
Enzymatic production of Cilastatin intermediate via highly enantioselective hydrolysis of methyl (+\/-)-2,2-dimethylcyclopropane carboxylate using newly isolated Rhodococcus sp. ECU1013
Applied Microbiology & Biotechnology 97 :7659

Liu CH, Pan J, Ye Q, Xu JH (2013)
Applied Microbiology & Biotechnology 97 :7659