Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, People's Republic of China.
Appl Microbiol Biotechnol. 2010 May;86(5):1493-501. doi: 10.1007/s00253-009-2382-0. Epub 2009 Dec 24.
Two alternative cell-surface display systems were developed in Pichia pastoris using the alpha-agglutinin and Flo1p (FS) anchor systems, respectively. Both the anchor cell wall proteins were obtained originally from Saccharomyces cerevisiae. Candida antarctica lipase B (CALB) was displayed functionally on the cell surface of P. pastoris using the anchor proteins alpha-agglutinin and FS. The activity of CALB displayed on P. pastoris was tenfold higher than that of S. cerevisiae. The hydrolytic and synthetic activities of CALB fused with alpha-agglutinin and FS anchored on P. pastoris were investigated. The hydrolytic activities of both lipases displayed on yeast cells surface were more than 200 U/g dry cell after 120 h of culture (200 and 270 U/g dry cell, respectively). However, the synthetic activity of CALB fused with alpha-agglutinin on P. pastoris was threefold higher than that of the FS fusion protein when applied to the synthesis of ethyl caproate. Similarly, the CALB displayed on P. pastoris using alpha-agglutinin had a higher catalytic efficiency with respect to the synthesis of other short-chain flavor esters than that displayed using the FS anchor. Interestingly, for some short-chain esters, the synthetic activity of displaying CALB fused with alpha-agglutinin on P. pastoris was even higher than that of the commercial CALB Novozyme 435.
分别利用α-凝集素和 Flo1p（FS）锚定系统在巴斯德毕赤酵母中开发了两种替代的细胞表面展示系统。这两种锚定细胞壁蛋白最初都来自酿酒酵母。利用锚定蛋白α-凝集素和 FS，将南极假丝酵母脂肪酶 B（CALB）功能性地展示在巴斯德毕赤酵母的细胞表面上。CALB 在巴斯德毕赤酵母上的展示活性比酿酒酵母高 10 倍。研究了与α-凝集素和 FS 融合并锚定在巴斯德毕赤酵母上的 CALB 的水解和合成活性。两种脂肪酶在酵母细胞表面的水解活性在培养 120 h 后均超过 200 U/g 干细胞（分别为 200 和 270 U/g 干细胞）。然而，当用于合成己酸乙酯时，CALB 与α-凝集素融合的合成活性是 FS 融合蛋白的三倍。同样，与 FS 锚定相比，利用α-凝集素在巴斯德毕赤酵母上展示的 CALB 在合成其他短链风味酯方面具有更高的催化效率。有趣的是，对于一些短链酯，利用α-凝集素在巴斯德毕赤酵母上展示融合的 CALB 的合成活性甚至高于商业 CALB Novozyme 435。
