Kleinbeck-Riniker, Florian, Schlama, Thierry, Ruch, Thomas, Novick, Scott, Dellas, Nikki, Garcia, Ravi, Ching, Charlene, Bautista, Abby, Homan, David, Alvizo, Oscar and Entwistle, David (2021) Engineering an omega-Transaminase for the Efficient Production of a Chiral Sacubitril Precursor. ACS Catalysis, 11. pp. 3762-3770. ISSN 2155-54352155-5435

Abstract

An omega-transaminase was engineered for the efficient production of a chiral precursor to sacubitril, (2R,4S)-5-([1,1'-biphenyl]-4-yl)-4-amino-2-methylpentanoic acid, a key component in the blockbuster heart failure drug Entresto®. Starting from an enzyme with trace activity and preference for the undesired diastereoisomer, eleven rounds of enzyme evolution were performed. The resultant variant, CDX-043, showed high productivity giving 90% conversion at 75 g/L substrate concentration with 1% enzyme loading with respect to the substrate in 24 h and without the use of an organic co-solvent. The product diastereomeric purity towards the desired (2R,4S)-stereoisomer was > 99:1 d.r. This variant also exhibited high process robustness and could tolerate reaction temperatures up to 65 °C and isopropylamine concentrations of at least 2 M. A structural analysis of the enzyme variants gave insight into how the mutations affected activity and selectivity. This new enzyme variant allows for the efficient and cost-effective production of sacubitril at large scale.

Item Type:	Article
Keywords:	amine transaminase, enzyme engineering, chiral amine, biocatalysis, green chemistry, sacubitril
Date Deposited:	24 Mar 2021 00:45
Last Modified:	24 Mar 2021 00:45
URI:	https://oak.novartis.com/id/eprint/41890