title: Protein engineering of nitrilase for chemoenzymatic production of glycolic acid.
abstract: A key step in a chemoenzymatic process for the production of high-purity glycolic acid (GLA) is the enzymatic conversion of glycolonitrile (GLN) to ammonium glycolate using a nitrilase derived from Acidovorax facilis 72W. Protein engineering and over-expression of this nitrilase, combined with optimized fermentation of an E. coli transformant were used to increase the enzyme-specific activity up to 15-fold and the biocatalyst-specific activity up to 125-fold. These improvements enabled achievement of the desired volumetric productivity and biocatalyst productivity for the conversion of GLN to ammonium glycolate.
authors.affiliation: Central Research and Development Department, E.I. du Pont de Nemours and Co., Experimental Station, P.O. Box 80328, Wilmington, Delaware 19880-0328, USA.
authors.forename: Shijun
authors.initials: S
authors.lastname: Wu
authors.forename: Arthur J
authors.initials: AJ
authors.lastname: Fogiel
authors.forename: Kelly L
authors.initials: KL
authors.lastname: Petrillo
authors.forename: Raymond E
authors.initials: RE
authors.lastname: Jackson
authors.forename: Kimberley N
authors.initials: KN
authors.lastname: Parker
authors.forename: Robert
authors.initials: R
authors.lastname: Dicosimo
authors.forename: Arie
authors.initials: A
authors.lastname: Ben-Bassat
authors.forename: Daniel P
authors.initials: DP
authors.lastname: O'Keefe
authors.forename: Mark S
authors.initials: MS
authors.lastname: Payne
mesh_terms: D000097:Acetonitriles / Q000737:chemistry*; D000619:Aminohydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D020562:Betaproteobacteria / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D006016:Glycolates / Q000138:chemical synthesis*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism