Novel (S)-Selective Hydrolase from Arthrobacter sp. K5 for Kinetic Resolution of Cyclic Amines

Abstract

Chiral 2-methylpiperidine (2-MPI) is an important building block that has potential for applications in pharmaceuticals and pesticides. In this study, we observed that the hydrolase in Arthrobacter sp. K5 exhibits high (S)-selectivity toward rac-N-pivaloyl-2-MPI to yield (S)-2-MPI with 80.2% enantiomeric excess (ee) in a 38.2% conversion. The hydrolase, which was identified by analyses of partial amino acid sequences of the purified enzyme and genome sequence of Arthrobacter sp. K5, exhibited moderate homology with amidohydrolases up to 67% (molinate hydrolase from Gulosibacter molinativorax). The hydrolase gene was overexpressed in Rhodococcus erythropolis. The recombinant cells produced (S)-2-MPI with 83.5% ee in a 48.4% conversion (E = 26.3) from 100 mM rac-N-pivaloyl-2-MPI. These results suggest the possibility of an efficient preparation of chiral 2-MPI in kinetic resolution.

Development of the Synthetic Route to PF-06878031 Part 2: Amide Reduction Route

“…The Option D resolution of the racemic amide 16 rac was investigated in parallel to the ester resolution work to assess if a higher optical purity could be obtained using this approach. 10 The hydrolase enzyme collection was used to screen the transformation and revealed that the hydrolysis of the amide functional group was extremely selective, whereas selective hydrolysis of the methyl ester was challenging. The majority of enzymes showed little to no activity; however, five enzymes appeared to be selective hydrolyzing the amide bond of the undesired R-enantiomer.…”

Bio-Approach for Obtaining Enantiomerically Pure Clopidogrel with the Use of Ionic Liquids

“…A high selectivity, more favorable reaction conditions, and biocompatibility are just a few of the positive characteristics that make enzyme-mediated transformations so valued as a potent alternative instrument in organic synthesis. Since organic synthesis relies on performing a stereoselective biotransformation, and is therefore significantly less expensive than using a “chiral pool,” kinetic resolution (racemate resolution) with the use of enzymes is one of the most widely used techniques [ 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 ]. It also avoids the use of environmentally hazardous and toxic chemical compounds.…”

Improvement of (S)-selective imine reductase GF3546 for the synthesis of chiral cyclic amines

“…To reduce the stress elicited by a high-concentration substrate on SIR46, we used Rhodococcus erythropolis cells as robust host cells for the construction of the expression system. 19 Recombinant R. erythropolis expressing the SIR46 gene successfully led an efficient reaction, achieving full conversion of 100 mM 2-MPN (Fig. S11, ESI†).…”