Protein engineering of nitrilase for chemoenzymatic production of glycolic acid.
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.
Wu
Shijun
S
Central Research and Development Department, E.I. du Pont de Nemours and Co., Experimental Station, P.O. Box 80328, Wilmington, Delaware 19880-0328, USA.
Fogiel
Arthur J
AJ
Petrillo
Kelly L
KL
Jackson
Raymond E
RE
Parker
Kimberley N
KN
Dicosimo
Robert
R
Ben-Bassat
Arie
A
O'Keefe
Daniel P
DP
Payne
Mark S
MS
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