CELLULASE AEC3, CELLULASE GENE-CONTAINING DNA FRAGMENT AND PRODUCTION OF CELLULASE AEC3

摘要

PURPOSE: To obtain a cellulase neutral in its optimal pH with carboxymethylcellulose as substrate, having each specific molecular weight and amino acid sequence, stable in alkaline region, useful for clothing detergents, sewage treatment agent, etc. ;CONSTITUTION: Aloemonas No.212 strain is inoculated into an alkaline medium (pH 9) and put to shaking culture at 30°C for a day, and the resulting microbes are collected by centrifugation. From the microbes, a chromosomal DNA is isolated by a conventional method and put to restriction enzyme treatment and then bound to a vecter to form into a recombinant DNA, which is then incorporated into Escherichia coli to carry out transformation. The resultant transformant is cloned to select cellulase-productive strain, from which plasmid is recovered and put to restriction enzyme treatment to make a subcloning and incorporation into Escherichia coli to effect manifestation, thus obtaining the objective cellulase AEC3, which has the following characteristics: (1) optimal pH: 7 with carboxymethylcellulose as substrate; (2) molecular weight: 68000; (3) having an amino acid sequence of e.g. the formula; and (4) the N-terminated amino acid for the matured protein is the 31st alanine.;COPYRIGHT: (C)1993,JPO&Japio