title: Protein engineering of Acidovorax facilis 72W nitrilase for bioprocess development.
abstract: Hydroxycarboxylic acid monomers can be used to prepare industrially important polymers. Enzymatic production of such hydroxycarboxylic acids is often preferred to chemical production since the reactions are run at ambient temperature, do not require strongly acidic or basic reaction conditions, and produce the desired product with high selectivity at high conversion. However, native enzymes often do not perform desired reactions with the efficiency required for commercial applications. Protein engineering was used to significantly increase the specific activity of nitrilase from Acidovorax facilis 72W for the conversion of 3-hydroxyvaleronitrile to 3-hydroxyvaleric acid. Overexpression of engineered nitrilase enzymes in Escherichia coli, combined with immobilization of whole cells in alginate beads that can be recycled many times has facilitated the development of a commercially viable bioprocess for production of 3-hydroxyvaleric acid.
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: Eugenia C
authors.initials: EC
authors.lastname: Hann
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Mersinger
authors.forename: Robert
authors.initials: R
authors.lastname: DiCosimo
authors.forename: Daniel P
authors.initials: DP
authors.lastname: O'Keefe
authors.forename: Arie
authors.initials: A
authors.lastname: Ben-Bassat
authors.forename: Mark S
authors.initials: MS
authors.lastname: Payne
mesh_terms: D000464:Alginates / Q000737:chemistry; D000619:Aminohydrolases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D019149:Bioreactors / Q000382:microbiology; D018914:Cells, Immobilized; D042621:Comamonadaceae / Q000201:enzymology*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005285:Fermentation; D007218:Industrial Microbiology; D008863:Microspheres; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering; D014170:Transformation, Genetic
chemical_list: D000464:Alginates; D000619:Aminohydrolases; C021867:nitrilase