Engineered nitrilases for biocatalysis

Abstract

The present invention provides an engineered nitrilase polypeptide capable of converting (1-cyanomethyl) cyclohexane-1-carbonitrile into (1-cyanocyclohexyl)-acetic acid. Utilizing advanced enzyme engineering techniques, the nitrilase exhibits enhanced stability and activity over natural variants. These engineered enzymes can hydrolyze a wide range of nitrile-containing compounds, including cyclic and aliphatic substrates. They handle higher substrate concentrations (200 g/L to 300 g/L), are thermostable above 50° C., and remain stable within a pH range of 5.5 to 8.0, making them suitable for various industrial applications. Their ability to convert substrates such as mandelonitrile, 2-(2-chlorophenyl)-2-hydroxyacetonitrile, 2-(6-methoxynaphthalen-2-yl)propanenitrile, and 2-[1-(aminomethyl)cyclohexyl]acetonitrile into corresponding carboxylic acids enables efficient and cost-effective production from diverse starting materials. This invention offers an engineered nitrilase enzyme as an alternative to alkaline or acid hydrolysis for converting nitrile substrates into carboxylic acids. It has applications in pharmaceuticals, agrochemicals, fine chemicals, waste treatment, and bioremediation, making these engineered polypeptides valuable for chemical production.

Dependencies are inferred from the claim text.