title: Chemoenzymatic synthesis of D(-)phenylglycine using hydantoinase of Pseudomonas desmolyticum resting cells.
abstract: We screened 125 Pseudomonas strains from our culture collection for the production of hydantoinase activity using DL-phenylhydantoin as a substrate. Pseudomonas desmolyticum NCIM 2112 was found to be the best hydantoinase (dihydropyrimidinase E.C. 3.5.2.2) producer. The enzymatic reactions were carried out using 18-20-h grown cells in nutrient broth and 5-phenylhydantoin as the substrate. Optimization studies for the biotransformation reaction were performed to increase product yield. The optimum pH and temperature for D(-)N-carbamoylphenylglycine production were 9.5 and 30 degrees C, respectively. Biotransformation under these alkaline conditions allowed the complete conversion of 27.0 g l-1 of DL-phenylhydantoin to 26.5 g l-1 of N-carbamoylphenylglycine within 24 h, with a molar yield of 90%. The hydantoinase involved in this biotransformation process was strictly D-stereospecific, because the product isolated was pure D(-)N-carbamoylphenylglycine. This pure product was further chemically converted to D(-)phenylglycine using nitrous acid with an 80% chemical yield. Thus, the overall conversion efficiency of DL-5-phenylhydantoin to D(-)phenylglycine was found to be 65-68%.
authors.affiliation: Division of Biochemical Sciences, National Chemical Laboratory, Pune, Maharashtra, India.
authors.forename: D V
authors.initials: DV
authors.lastname: Gokhale
authors.forename: K B
authors.initials: KB
authors.lastname: Bastawde
authors.forename: S G
authors.initials: SG
authors.lastname: Patil
authors.forename: U R
authors.initials: UR
authors.lastname: Kalkote
authors.forename: R R
authors.initials: RR
authors.lastname: Joshi
authors.forename: R A
authors.initials: RA
authors.lastname: Joshi
authors.forename: T
authors.initials: T
authors.lastname: Ravindranathan
authors.forename: B G
authors.initials: BG
authors.lastname: Gaikwad
authors.forename: V V
authors.initials: VV
authors.lastname: Jogdand
authors.forename: S
authors.initials: S
authors.lastname: Nene
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism*; D001639:Bicarbonates / Q000494:pharmacology; D001711:Biotransformation / Q000235:genetics; D001881:Borates / Q000494:pharmacology; D004151:Dipeptides / Q000138:chemical synthesis* / Q000302:isolation & purification; D005998:Glycine / Q000031:analogs & derivatives* / Q000302:isolation & purification; D006827:Hydantoins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D015394:Molecular Structure; D009608:Nitrous Acid / Q000378:metabolism; D010648:Phenylacetates / Q000302:isolation & purification / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology; D013237:Stereoisomerism; D013696:Temperature; D014508:Urea / Q000031:analogs & derivatives / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D001639:Bicarbonates; D001881:Borates; D004151:Dipeptides; D006827:Hydantoins; D010648:Phenylacetates; C096868:N-carbamyl(phenyl)glycine; C008852:2-phenylglycine; C098839:5-phenylhydantoin; D014508:Urea; D000581:Amidohydrolases; C020047:dihydropyrimidinase; D009608:Nitrous Acid; D005998:Glycine