GENE OF MALTOPENTAOSE-FORMING ENZYME, MICROORGANISM CONTAINING THE GENE, PRODUCTION OF MALTOPENTAOSE-FORMING ENZYME USING THE MICROORGANISM AND NOVEL ENZYME PRODUCED THEREBY

摘要

PURPOSE:To improve the productivity of maltopentaose-producing enzyme by integrating a cloned maltopentaose-producing enzyme gene into a vector, introducing the vector into a host microoganism and culturing the transformant. CONSTITUTION:DNA of a donor microorganism capable of producing maltopentaose-producing enzyme is incised with ultrasonic wave, restriction enzyme, etc., to obtain a vector fragment. Separately, another vector fragment is produced by incising a vector such as pBR322 by the similar method. Both vector fragments are linked together using a DNA ligase, etc., to obtain a recombinant DNA containing a maltopentaose-producing enzyme gene. The recombinant DNA is introduced into a host microorganism such as Escherichia coli, the obtained transformant is cultured in liquid medium and centrifuged, the separated microbial cells are disintegrated, the broken cells are removed and the remaining component is purified to obtain a maltopentaose enzyme having a molecular weight of 65,000 (SDS-polyacrylamide slab gel electrophoresis), an optimum pH of 6.0 and an optimum temperature of 37-45 deg.C.