Sagt C M, Kleizen B, Verwaal R, de Jong M D, Müller W H, Smits A, Visser C, Boonstra J, Verkleij A J, Verrips C T
Department of Molecular Cell Biology and the Institute of Biomembranes, Utrecht University, 3584 CH Utrecht, The Netherlands.
Appl Environ Microbiol. 2000 Nov;66(11):4940-4. doi: 10.1128/AEM.66.11.4940-4944.2000.
Saccharomyces cerevisiae is often used to produce heterologous proteins that are preferentially secreted to increase economic feasibility. We used N-glycosylation as a tool to enhance protein secretion. Secretion of cutinase, a lipase, and llama V(HH) antibody fragments by S. cerevisiae or Pichia pastoris improved following the introduction of an N-glycosylation site. When we introduced an N-glycosylation consensus sequence in the N-terminal region of a hydrophobic cutinase, secretion increased fivefold. If an N-glycosylation site was introduced in the C-terminal region, however, secretion increased only 1.8-fold. These results indicate that the use of N glycosylation can significantly enhance heterologous protein secretion.
酿酒酵母常用于生产优先分泌的异源蛋白，以提高经济可行性。我们使用N-糖基化作为增强蛋白分泌的工具。酿酒酵母或巴斯德毕赤酵母分泌角质酶、脂肪酶和骆驼V(HH)抗体片段的能力在引入N-糖基化位点后得到改善。当我们在疏水性角质酶的N端区域引入N-糖基化共有序列时，分泌量增加了五倍。然而，如果在C端区域引入N-糖基化位点，分泌量仅增加1.8倍。这些结果表明，使用N-糖基化可以显著增强异源蛋白的分泌。
