Development of new microbial enzymes and their application to organic synthesis

Abstract

We screened microorganisms for various new enzymes, which were used in the synthesis of organic chemicals. A new hydro-lyase from Arthrobacter sp. catalyzed the synthesis of 87 grams/liter of D-malate from maleic acid in 72% yield. A. denitrificans produced 27 g of (S)-(+)-citramalate of 99.9 % ee in 69% yield. Phenylalanine dehydrogenase (PheDH) is used in the colorimetric micro-determination of L-Phe to detect phenylketonuria (PKU) in the blood of neonates in Japan. The structure of a new enzyme opine dehydrogenase (ODH) from Arthrobacter sp., has been solved to 1.8 A resolution. Optically pure secondary amine carboxylic acids were synthesized by ODH. Picolinic acid derivatives were synthesized by Pseudomonas catechol 2,3-oxygenase. Nearly 20 grams/liter of theobromine was produced from caffeine in a yield of 92% by P. putida cells. A new enzyme alkaline D-peptidase from B. cereus was used for oligomerization of D-Phe methylester.