E.i. Dupont De Nemours and Company

Sarita Chauhan

Mark Scott Payne

John Edward Gavagan

Robert Donald Fallon

Robert Dicosimo

Robert DiCosimo

Michael G Bramucci

Leo Ernest Manzer

A method has been developed to prepare (E)- and (Z)-2-methyl-2-butenoic acids (2M2BA) from a mixture of (E,Z)-2-methyl-2-butenenitriles (2M2BN) by the regioselective hydrolysis of (E)-2M2BN to (E)-2-methyl-2-butenoic acid (2M2BA) using enzyme catalysts having either a nitrilase activity or a combination of nitrile hydratase and amidase activities. The method provides high yields without significant conversion of (Z)-2M2BN to (Z)-2M2BA. The regioselective hydrolysis of (E)-2M2BN to (E)-2M2BA makes possible the facile separation of (E)-2M2BA from (Z)-2M2BN or (Z)-2-methyl-2-butenamide (2M2BAm), and the subsequent conversion of (Z)-2M2BN or (Z)-2M2BAm to (Z)-2M2BA.

Preparation of (E)- and (Z)-2-methyl-2-butenoic acids

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Recombinant microbial strains are provided that express nitrilase enzyme and are useful as biocatalysts for the hydrolysis of nitrile-containing substrates. The recombinant cells are transformed with a foreign gene isolated from72W encoding a thermostable nitrilase enzyme that catalyzes the hydrolysis of nitrile-containing substrates to carboxylic acids under mild reaction conditions. The nucleotide sequence of the nitrilase gene and the deduced amino acid sequence encoded by the nitrilase gene are provided.

Isolation and expression of a gene for a nitrilase from Acidovorax facilis 72W

The present invention relates to a method for producing 2-hydroxyisobutyric acid where acetone cyanohydrin is converted to 2-hydroxyisobutyric acid using an enzyme catalyst having nitrilase activity, or having a combination of nitrile hydratase and amidase activities. The invention also encompasses production of methacrylic acid wherein the 2-hydroxyisobutyric acid produced with the catalyst described is dehydrated to produce methacrylic acid.

Method for producing 2-hydroxyisobutyric acid and methacrylic acid from acetone cyanohydrin

This invention relates to a process for preparing dicarboxylic acid monoesters from cyanocarboxylic acid esters. More particularly, using the nitrilase from 72W (ATCC 55745) in various forms as an enzyme catalyst, an aliphatic or aromatic cyanocarboxylic acid ester is converted to the corresponding dicarboxylic acid monoester with high chemoselectivity at 100% conversion. An embodiment of the invention also provides a process for obtaining high chemoselectively and high regioselectivity.

Preparation of dicarboxylic acid monoesters from cyanocarboxylic acid esters

The present invention relates to a method for producing &agr;-hydroxy acids using an enzyme catalyst having nitrilase activity. More specifically, the invention pertains to use of 72W (ATCC 55746) nitrilase to hydrolyze glycolonitrile to glycolic acid. Glycolonitrile is reacted in an aqueous mixture with a catalyst having 72W nitrilase activity to give glycolic acid selectively, and at high concentration and high yield.

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Landenberg, PA, United States of America

Sarita Chauhan