Display of fungal hydrophobin on the Pichia pastoris cell surface and its influence on Candida antarctica lipase B.
Display of fungal hydrophobin on the Pichia pastoris cell surface and its influence on Candida antarctica lipase B.
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毕赤酵母细胞表面真菌疏水蛋白的展示及其对南极假丝酵母脂肪酶B的影响
DOI：
10.1007/s00253-016-7431-x
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发表时间：
2016-07
影响因子：
5
通讯作者：
Lin Y
中科院分区：
文献类型：
--
作者：
Wang P;He J;Sun Y;Reynolds M;Zhang L;Han S;Liang S;Sui H;Lin Y
为了修饰毕赤酵母细胞表面，将裂褶菌SC 3和里氏木霉HFBI两类疏水蛋白分别展示在细胞壁上。重组菌株的疏水性明显增加，并在修饰细胞上共展示了抗真菌假丝酵母酶B（CALB），得到了GS 115/SC 3 -61/CALB-51和GS 115/HFBI-61/CALB-51两株重组菌株。有趣的是，与减疏水蛋白重组菌株GS 115/CALB-51相比，菌株GS 115/HFBI-61/CALB-51的水解和合成活性分别增加了37%和109%，但菌株GS 115/SC 3 -61/CALB-51的水解和合成活性分别下降了26%和43%。疏水蛋白修饰后，吸附在细胞表面上的酯交换反应的甘油副产物的量显著降低，去除甘油屏障，并允许底物进入脂肪酶的活性位点。电子显微镜观察表明，两种重组菌株的细胞壁结构发生了变化，包括内部葡聚糖层和外部甘露聚糖层的变化。这些结果表明疏水蛋白的展示可以改变P. pastoris对CALB催化活性的影响。巴斯德氏细胞
To modify thePichia pastoriscell surface, two classes of hydrophobins, SC3 fromSchizophyllum communeand HFBI fromTrichoderma reesei, were separately displayed on the cell wall. There was an observable increase in the hydrophobicity of recombinant strains.Candida antarcticalipase B (CALB) was then co-displayed on the modified cells, generating strains GS115/SC3–61/CALB-51 and GS115/HFBI-61/CALB-51. Interestingly, the hydrolytic and synthetic activities of strain GS115/HFBI-61/CALB-51 increased by 37 and 109 %, respectively, but decreased by 26 and 43 %, respectively, in strain GS115/SC3–61/CALB-51 compared with the hydrophobin-minus recombinant strain GS115/CALB-GCW51. The amount of glycerol by-product from the transesterification reaction adsorbed on the cell surface was significantly decreased following hydrophobin modification, removing the glycerol barrier and allowing substrates to access the active sites of lipases. Electron micrographs indicated that the cell wall structures of both recombinant strains appeared altered, including changes to the inner glucan layer and outer mannan layer. These results suggest that the display of hydrophobins can change the surface structure and hydrophobic properties ofP. pastorisand affect the catalytic activities of CALB displayed on the surface ofP. pastoriscells.