Chemoenzymatic synthesis of D(-)phenylglycine using hydantoinase of Pseudomonas desmolyticum resting cells.
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%.
Gokhale
D V
DV
Division of Biochemical Sciences, National Chemical Laboratory, Pune, Maharashtra, India.
Bastawde
K B
KB
Patil
S G
SG
Kalkote
U R
UR
Joshi
R R
RR
Joshi
R A
RA
Ravindranathan
T
T
Gaikwad
B G
BG
Jogdand
V V
VV
Nene
S
S
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
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