Tokunaga M, Kawamura A, Omori A, Hishinuma F
Laboratory of Molecular Genetics, Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Biochim Biophys Acta. 1991 Oct 25;1080(2):135-7. doi: 10.1016/0167-4838(91)90139-q.
We have previously reported the construction of recombinant mouse salivary alpha-amylase secretion vector in Saccharomyces cerevisiae utilizing novel yeast secretion signal derived from killer 28 kDa precursor protein. Here, we have first purified recombinant mouse alpha-amylase to homogeneity from the culture medium of S. cerevisiae, and determined its NH2-terminal amino acid sequence. The sequencing data indicated that the 28 kDa killer secretion signal-alpha-amylase fusion protein was cleaved accurately at its native processing site, and that both the core-glycosylated and non-glycosylated alpha-amylases possessed the same NH2-terminal amino acid sequences.
我们之前报道过，利用源自杀伤性28 kDa前体蛋白的新型酵母分泌信号，在酿酒酵母中构建重组小鼠唾液α-淀粉酶分泌载体。在此，我们首次从酿酒酵母培养基中纯化出了均一的重组小鼠α-淀粉酶，并测定了其NH2末端氨基酸序列。测序数据表明，28 kDa杀伤性分泌信号-α-淀粉酶融合蛋白在其天然加工位点被准确切割，并且核心糖基化和非糖基化的α-淀粉酶都具有相同的NH2末端氨基酸序列。
