Yi, Xiang, Kleinbeck-Riniker, Florian, Chang, Charlene, Boghospor, Lorita, Gomes, Sandy, Alvizo, Oscar, Allmendinger, Thomas, Fell, Jason, Subramanian, Nandhitha, Li, Michelle, Garcia, Ravi, Riggins, James, Entwistle, David, Richter, Yvonne, Gschwend, Daniel, Lauener, Liam, Schlama, Thierry, Ruch, Thomas, Peat, Thomas S. and Lebhar, Helene (2024) Enhancing the imine reductase activity of a promiscuous glucose dehydrogenase for scalable manufacturing of a chiral neprilysin inhibitor precursor. ACS Catalysis, 14 (9). pp. 7087-7096. ISSN 2155-54352155-5435

Abstract

Imine reductases (IREDs) have been identified as an important class of biocatalysts to synthesize chiral secondary amines with substantial promise for industrial application. Here, we report the promiscuous imine reductase activity of a glucose dehydrogenase (GDH), representing a new class of highly selective IREDs. Starting from GDH-105, a commercial glucose dehydrogenase variant typically used for NAD(P)H regeneration, eight rounds of directed evolution were used to convert this enzyme into a highly active IRED for the manufacture of a chiral neprilysin inhibitor precursor with excellent chemo- and stereoselectivity, improved NADH cofactor specificity, and high thermal stability. The evolved variant GDH-201 showed excellent productivity of 99% conversion over 3 h at 50 g/L keto substrate concentration and 10% enzyme loading with respect to the keto substrate. Early process development studies at multigram scale provided the product in 94% yield with >99% purity as a single diastereomer.

Item Type:	Article
Keywords:	Imine reductase, enzyme evolution, neprilysin inhibitor
Date Deposited:	18 May 2024 00:46
Last Modified:	18 May 2024 00:46
URI:	https://oak.novartis.com/id/eprint/51795