利用 Cwp2 作为锚定蛋白在酿酒酵母中表面展示活性脂肪酶。
Surface display of active lipase in Saccharomyces cerevisiae using Cwp2 as an anchor protein.
机构信息
Key Laboratory of Molecular Biophysics, The Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, 430074 Wuhan, China.
出版信息
Biotechnol Lett. 2010 Feb;32(2):255-60. doi: 10.1007/s10529-009-0138-7. Epub 2009 Oct 11.
Lipase Lip2 from Yarrowia lipolytica was displayed on the cell surface of Saccharomyces cerevisiae using Cwp2 as an anchor protein. Successful display of the lipase on the cell surface was confirmed by immunofluorescence microscopy and halo assay. The length of linker sequences was further examined to confirm that the correct conformation of Lip2 was maintained. The results showed that the displayed Lip2 exhibited the highest activity at 7.6 +/- 0.4 U/g (dry cell) when using (G(4)S)(3) sequence as the linker, with an optimal temperature and pH at 40 degrees C and pH 8.0. The displayed lipase did not lose any activity after being treated with 0.1% Triton X-100 and 0.1% Tween 80 for 30 min, and it retained 92% of its original activity after incubation in 10% DMSO for 30 min. It also exhibited better thermostability than free Lip2 as reported previously.
脂肪酶 Lip2 来自假丝酵母利用 Cwp2 作为锚定蛋白展示在酿酒酵母的细胞表面。通过免疫荧光显微镜和晕圈试验证实了脂肪酶在细胞表面的成功展示。进一步检查连接序列的长度以确认 Lip2 的正确构象得以维持。结果表明，当使用 (G(4)S)(3) 序列作为连接子时，展示的 Lip2 在 7.6 +/- 0.4 U/g（干细胞）时表现出最高的活性，最适温度和 pH 值分别为 40 摄氏度和 pH 8.0。在 0.1% Triton X-100 和 0.1% Tween 80 处理 30 分钟后，展示的脂肪酶没有失去任何活性，在 10% DMSO 孵育 30 分钟后保留了 92%的原始活性。与之前报道的游离 Lip2 相比，它还表现出更好的热稳定性。
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