Protein design on pyruvate decarboxylase (PDC) by site-directed mutagenesis. Application to mechanistical investigations, and tailoring PDC for the use in organic synthesis.
Pyruvate decarboxylases (E.C. 4.1.1.1) from various organisms have been studied for many years, mainly with respect to the mechanism of the non-oxidative decarboxylation reaction. Although the C-C-bond-forming properties of these enzymes are known and have been applied for many years in biotransformations for the synthesis of chiral alpha-hydroxy ketones, only little is known about the factors influencing the carboligase side-reaction. The present review surveys recent efforts in the study of site-directed mutagenesis on PDCs, which are discussed against a background of the structural and kinetical investigations. It also includes recent studies on tailoring the PDCs of Zymomonas mobilis for the syntheses of (R)-phenylacetyl carbinol (PAC), a pre-step in the synthesis of L-ephedrine, by protein design techniques.
Pohl
M
M
Institut für Enzymtechnologie, Heinrich-Heine Universität Düsseldorf, im Forschungszentrum Jülich.
D000595:Amino Acid Sequence; D001711:Biotransformation; D017076:Computer-Aided Design; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011767:Pyruvate Decarboxylase / Q000737:chemistry* / Q000235:genetics; D016415:Sequence Alignment
D007527:Isoenzymes; D011767:Pyruvate Decarboxylase