Ruohonen L, Hackman P, Lehtovaara P, Knowles J K, Keränen S
Recombinant DNA Laboratory, University of Helsinki, Finland.
Gene. 1987;59(2-3):161-70. doi: 10.1016/0378-1119(87)90324-6.
The expression and secretion of Bacillus amyloliquefaciens alpha-amylase was studied in yeast Saccharomyces cerevisiae. The Bacillus promoter was removed by BAL 31 digestion and three forms of the alpha-amylase gene were constructed: the Bacillus signal sequence was either complete (YEp alpha a1), partial (YEp alpha a2) or missing (YEp alpha a3). Secretion of alpha-amylase into the culture medium was obtained with the complete signal sequence only. The secreted alpha-amylase was glycosylated and its signal peptide was apparently processed. The glycosylated alpha-amylase remained active. The enzyme produced by the other constructions was not glycosylated and thus probably remained in the cytoplasm.
在酿酒酵母中研究了解淀粉芽孢杆菌α-淀粉酶的表达和分泌。通过BAL 31酶切去除芽孢杆菌启动子，并构建了三种形式的α-淀粉酶基因：芽孢杆菌信号序列完整（YEpαa1）、部分完整（YEpαa2）或缺失（YEpαa3）。仅在信号序列完整时，α-淀粉酶才能分泌到培养基中。分泌的α-淀粉酶进行了糖基化，其信号肽明显被加工。糖基化的α-淀粉酶仍保持活性。其他构建体产生的酶未进行糖基化，因此可能留在细胞质中。
