title: The stereoselectivity and hydrolysis efficiency of recombinant D-hydantoinase from Vigna angularis Against 5-benzylhydantoin derivatives with halogen and methyl substituents.
abstract: The researches on D-hydantoinase activity and substrate specificity towards dihydropyrimidine and hydantoin derivatives have been carried out intensively over the last few decades. So far, the major efforts have focused on (R,S)-5-phenylhydantoin and (R,S)-5-(4-hydroxyphenyl)hydantoin, the most desirable D-hydantoinase substrates from pharmaceutical industry point of view. However, it was shown that D-hydantoinase is a substrate-dependent enzyme, and its activity and stereoselectivity towards 5-monosubstituted hydantoins varied significantly with the type of substrate and the source of the enzyme. The aim of this study was to estimate the substrate specificity of D-hydantoinase towards series of 5-benzylhydantoin derivatives with halogen and methyl substituents in the phenyl ring. The biotransformations were carried out by using commercial enzyme: immobilized, recombinant, cloned, and expressed in Escherichia coli D-hydantoinase from Vigna angularis (rD-HYD). All reactions were monitored by capillary electrophoresis (CE), and the conversion yields were calculated. Additionally, enantiomeric ratios of the obtained D-phenylalanine derivatives were estimated by chiral high-performance liquid chromatography (HPLC). Interestingly, the differences in the activities of examined enzyme towards particular 5-benzylhydantoin derivatives were observed. CE was also shown as a promising method for monitoring the hydrolysis of new substrates by D-hydantoinase and further analyzing of enzyme substrate specificity.
authors.affiliation: Department of Technology and Biotechnology of Drugs, Jagiellonian University Medical College, Medyczna 9, 30-688, Kraków, Poland, glatacz@cm-uj.krakow.pl.
authors.forename: Gniewomir
authors.initials: G
authors.lastname: Latacz
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kieć-Kononowicz
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D019075:Electrophoresis, Capillary; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007887:Fabaceae / Q000737:chemistry* / Q000201:enzymology; D015870:Gene Expression; D006219:Halogens / Q000737:chemistry; D006827:Hydantoins / Q000737:chemistry*; D006868:Hydrolysis; D010649:Phenylalanine / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013237:Stereoisomerism; D013379:Substrate Specificity; D014169:Transformation, Bacterial
chemical_list: D006219:Halogens; D006827:Hydantoins; D010940:Plant Proteins; D011994:Recombinant Proteins; C031427:5-benzylhydantoin; D010649:Phenylalanine; C098839:5-phenylhydantoin; D000581:Amidohydrolases; C020047:dihydropyrimidinase