HEAT-RESISTANT TREHALOSE PHOSPHORYLASE, ITS PRODUCTION, BACTERUM TO BE USED FOR ITS PRODUCTION, AND PRODUCTION OF TREHALOSE USING THE ENZYME

摘要

PURPOSE: To obtain the subject new enzyme active at elevated temperatures, thus capable of giving trehalose in an industrially advantageous way in a short reaction time with reduced sundry germ contamination, by culturing trehalose phosphorylase-productive microbes belonging to the genus Bacillus. CONSTITUTION: Trehalose phosphorylase-productive microbes belonging to the genus Bacillus [e.g. Bacillus stearothermophilus SK-1 (FERM P-14567)] is inoculated into a medium and subjected to aerating agitation culture at 55 deg.C for 72hr followed by centrifugal separation to take out the supernatant; ammonium sulfate is then dissolved in the supernatant so as to come to 40-60% in the degree of saturation, and the resultant protein precipitate is recovered by centrifugal separation and then dissolved in a 10mM potassium phosphate-citric acid buffer solution (pH6.0) followed by purification through e.g. ion exchange chromatography, hydrophobic chromatography, etc., thus obtaining the objective new heat-resistant trehalose phosphorylase retaining >=95% activity after treated in a pH6.0 buffer solution at 50-65 deg.C for 15min and capable of producing trehalose in an industrially advantageous way.