Tokunaga M, Kawamura A, Yonekyu S, Kishida M, Hishinuma F
Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Yeast. 1993 Apr;9(4):379-87. doi: 10.1002/yea.320090408.
We have constructed two secretion vectors for Schizosaccharomyces pombe using an SV40 promoter and the secretion signals of the pGKL killer toxin complex derived from Kluyveromyces lactis. Although indigenous secretory glycoproteins tend to accumulate in the periplasmic space of S. pombe, we have succeeded in the secretion of mouse alpha-amylase into the culture medium. The efficiency of secretion, processing pattern, stability and culture conditions for mouse alpha-amylase were studied in S. pombe. The 128 kDa killer secretion signal was more effective in directing secretion of mouse alpha-amylase than the 28 kDa killer secretion signal. We detected a chymostatin-sensitive protease activity in the culture medium of S. pombe, which digests mouse alpha-amylase secreted into the culture medium. The addition of 5 micrograms/ml chymostatin was shown to protect mouse alpha-amylases from this degradation.
我们使用SV40启动子和源自乳酸克鲁维酵母的pGKL杀伤毒素复合物的分泌信号构建了两种用于粟酒裂殖酵母的分泌载体。尽管本地分泌型糖蛋白倾向于在粟酒裂殖酵母的周质空间中积累，但我们已成功将小鼠α-淀粉酶分泌到培养基中。在粟酒裂殖酵母中研究了小鼠α-淀粉酶的分泌效率、加工模式、稳定性和培养条件。128 kDa的杀伤分泌信号在指导小鼠α-淀粉酶分泌方面比28 kDa的杀伤分泌信号更有效。我们在粟酒裂殖酵母的培养基中检测到一种糜蛋白酶敏感的蛋白酶活性，它能消化分泌到培养基中的小鼠α-淀粉酶。结果表明，添加5微克/毫升的糜蛋白酶抑制剂可保护小鼠α-淀粉酶不被降解。
