Cino P M, Tewari R P
Can J Microbiol. 1975 Sep;21(9):1362-8. doi: 10.1139/m75-204.
The physiochemical characteristics of the intracellular proteolytic enzymes of Oidiodendron kalari, a neuropathogenic fungus, were studied. The organism in the yeast phase was grown in a semisynthetic medium containing 1% tryptone, at 37 degrees C for 48 hr, on a gyrotory shaker. The crude extract was prepared by breaking the cells in a French pressure cell and the proteolytci activity was tested against biological substrates. The cell-free extract hydrolyzed casein, hemoglobin, lactalbumin, gelatin, elastin, collagen and purified rabbit renal basement membrane to various degrees. Optimal proteolytic activity was observed at pH 6 and at 32 degrees C. Calcium and EDTA did not affect the enzymatic activity; however, activity was partially inhibited by sulfhydryl-blocking agents and by heat-inactivated horse, calf, and human serum. The extract was totally inactivated by exposure to a temperature of 70 degrees C for 60 min. Storage at -76 degrees C or -15 degrees C for 6 months or at 4 degrees C for 4 weeks did not affect protease activity.
对神经致病真菌卡拉里拟内孢霉细胞内蛋白水解酶的理化特性进行了研究。酵母阶段的该微生物在含有1%胰蛋白胨的半合成培养基中,于37℃在旋转摇床上培养48小时。通过在法国压榨器中破碎细胞制备粗提物,并针对生物底物测试蛋白水解活性。无细胞提取物对酪蛋白、血红蛋白、乳白蛋白、明胶、弹性蛋白、胶原蛋白和纯化的兔肾基底膜有不同程度的水解作用。在pH 6和32℃时观察到最佳蛋白水解活性。钙和乙二胺四乙酸不影响酶活性;然而,活性受到巯基阻断剂以及热灭活的马、小牛和人血清的部分抑制。提取物在70℃暴露60分钟后完全失活。在-76℃或-15℃储存6个月或在4℃储存4周不影响蛋白酶活性。