| 摘要 |
PURPOSE: To obtain the subject new enzyme having specific thermal stability, useful as e.g. a stable liquefied clinical diagnostic agent for glyceride or lipase activity assay, by culturing glyserol kinase-producing bacteria belonging to Flavobacterium. CONSTITUTION: Glycerol kinase-producing bacterum belonging to Flavobacterium [e.g. Flavobacterium meningosepticum GK54 (FERM BP-4868) strain] is subjected to seed culture and then transferred to a defoaming agent-contg. medium in a jar fermenter, where the microbes are cultured at 28 deg.C for 22hr the resultant cultured product is centrifuged to recover the microbes, which are then suspended in a 20mM tris-hydrochloric acid buffer solution (pH8.6) to which lysozyme is added to effect bacteriolysis; the resultant system is centriguged and the supernatant formed is purified by e.g. ion exchange chromatography, hydrophobic chromatographpy, etc., thus obtaining the objective new glycerol kinase retaining >=80% of its activity even after treated at 70 deg.C for 10min, useful for e.g. assaying glyceride or lipase activity and enabling to be used as a highly thermally stable liquefied reagent. |
