HEAT-RESISTANT BRANCHING ENZYME GENE, RECOMBINED PLASMID CONTAINING THE SAME AND HEAT-RESISTANT BRANCHING ENZYME

摘要

PURPOSE: To provide the subject novel enzyme gene originated from a thermophilic bacterium and enabling the high yield production of a heat-resistant branching enzyme capable of decomposing starch at high temperatures preventing the contamination of various bacteria. ;CONSTITUTION: A novel heat-resistant branching enzyme capable of decomposing starch at temperatures above 50°C is obtained by inoculating a medium with a thermophilic bacterium, Bacillus stearothtermophilus TRBE 14 strain (FERM P-13916), culturing the bacterium at 50°C for a night, centrifugally collecting the cultured bacterium cells, separating the chromosomal DNA of the bacterium cells by a phenol method, treating the DNA with a restriction enzyme, binding the obtained fragment to a phage vector, introducing the obtained recombined λ-phage particle into Escherichia coli, screening the transformed Escherichia coli with a probe comprising a part of a branching enzyme gene, recovering DNA from the screened clone of a heat-resistant branching enzyme gene, treating the DNA with a restriction enzyme, binding the obtained gene fragment to a vector, and subsequently expressing the obtained recombined plasmid in host cells.;COPYRIGHT: (C)1995,JPO