| Title : Enzymes for the resolution of alpha-tertiary-substituted carboxylic acid esters - Kallwass_1994_Bioorg.Med.Chem_2_557 |
| Author(s) : Kallwass HK , Yee C , Blythe TA , McNabb TJ , Rogers EE , Shames SL |
| Ref : Bioorganic & Medicinal Chemistry , 2 :557 , 1994 |
| Abstract : Aromatic alpha-amino-alpha-methyl acids and alpha-hydrazino-alpha-methyl acids are known aromatic amino acid decarboxylase inhibitors. Specific derivatives such as 2-amino-2-methyl-3-(3,4- dihydroxyphenyl)propanoate, Aldomet, and 2-hydrazino-2-methyl-3-(3,4- dihydroxyphenyl)propanoate, Lodosyn, have been developed as therapeutic agents to treat hypertension and Parkinson's disease, respectively. We recently reported a method for the kinetic resolution of the racemic esters of such compounds using a crude preparation of a novel enzyme catalyst from the yeast Candida lipolytica (Yee, C.; Blythe, T.A., McNabb, T.J.; Walts, A.E. J. Org. Chem. 1992, 57, 3525-3527). Here we report the purification and initial characterization of the active enzyme component, an enzyme given the name Candida lipolytica ester hydrolase (CLEH). CLEH was purified to > 95% homogeneity by chromatography on Matrex Blue B resin. The enzyme was found to be a glycoprotein with M(r) = 80,000-300,000. In addition to esterolytic activity, the enzyme was found to catalyze the hydrolysis of amides, anilides and peptides. Sequence analysis of internal peptides of CLEH revealed striking homology to a number of enzymes belonging to the group of serine carboxypeptidases (E.C. 3.4.16.1). One peptide aligned with the canonical serine carboxypeptidase active site sequence, GESYAG. Based on the structural relationship of CLEH to serine carboxypeptidases, three representative serine carboxypeptidases were evaluated for their utility in resolving racemic alpha-tertiary ester substrates and compared with the activity of CLEH. All enzymes revealed similarly high activity and enantioselectivity towards the alpha-hydrazino-alpha-methyl ester precursor of the Parkinson-drug Carbidopa. However, differences in enantioselectivity were observed with other alpha-tertiary-substituted ester substrates. Serine carboxypeptidase-catalyzed ester resolutions thus offer a new route to many sterically hindered homochiral alpha-amino, alpha-hydrazino and alpha-hydroxy carboxylic acids. |
| ESTHER : Kallwass_1994_Bioorg.Med.Chem_2_557 |
| PubMedSearch : Kallwass_1994_Bioorg.Med.Chem_2_557 |
| PubMedID: 7858960 |
Kallwass HK, Yee C, Blythe TA, McNabb TJ, Rogers EE, Shames SL (1994)
Enzymes for the resolution of alpha-tertiary-substituted carboxylic acid esters
Bioorganic & Medicinal Chemistry
2 :557
Kallwass HK, Yee C, Blythe TA, McNabb TJ, Rogers EE, Shames SL (1994)
Bioorganic & Medicinal Chemistry
2 :557