RECOMBINANT PLASMID HAVING GENE OF ENZYME ASYMMETRICALLY HYDROLYZING CARBOXYLIC ACID ESTER, MICROORGANISM TRANSFORMED THEREWITH AND PRODUCTION OF OPTICALLY ACTIVE CARBOXYLIC ACID ESTER WITH SAID MICROORGANISM

摘要

PURPOSE:To produce an optically active carboxylic acid, by inserting an esterase gene of Pseudomonas.fluorescens into plasmids pUC18 and pUC19, introducing the resultant plasmids into Escherichia coli to provide an esterase and using the above- mentioned enzyme. CONSTITUTION:A chromosomic DNA of Pseudomonas.fluorescens IFO3081 is digested with a restriction enzyme ECoRI, then linked to pUC18 similarly treated with the ECoRI using a T4 ligase and introduced into Escherichia coli JM109 strain to provide a plasmid pFY501 from a transformed strain. A plasmid pFY513 having an SmaI fragment having 2.2kb of a HindIII fragment having 4.9kb of the above-mentioned plasmid is then prepared. The afore-mentioned recombinant plasmids are subsequently introduced into a host microorganism (preferably Escherichia coli). The resultant culture fluid, microbial cells or treated substances thereof are reacted with a carboxylic acid ester expressed by formula I (R1 is alkyl, aralkyl or aryl; R2 and R3 are alkyl; n is 1 or 2) to provide an optically active carboxylic acid expressed by formula II.