The diversity of glycosylation of cellobiohydrolase I from Trichoderma reesei determined with mass spectrometry.
The diversity of glycosylation of cellobiohydrolase I from Trichoderma reesei determined with mass spectrometry.
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DOI：
10.1016/j.bbrc.2018.12.013
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发表时间：
2019-01
影响因子：
3.1
通讯作者：
Mingyu Wang;Ya-Nan Ma;Ling Li;Bianfang Wang;Xin Wei;Mengge Zhang;Juan Wang;Qingyu Cui;Zhiqiang Li;Hai Xu
中科院分区：
文献类型：
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作者：
Mingyu Wang;Ya-Nan Ma;Ling Li;Bianfang Wang;Xin Wei;Mengge Zhang;Juan Wang;Qingyu Cui;Zhiqiang Li;Hai Xu
Cellulases are glycosylated enzymes that have wide applications in fields like biofuels. It has been widely accepted that glycosylation of cellulases impact their performance.Trichoderma reeseiis the most important cellulase-producer and cellobiohydrolase I (CBHI) is the most important cellulase fromT. reesei. Therefore, the glycosylation ofT. reeseiCBHI has been a focus of research. However, investigations have been focused onN-glycosylation of three of the four potential glycosylation sites, as well asO-glycosylation on the linker region, while a full picture of glycosylation ofT. reeseiCBHI is still needed. In this work, with extensive mass spectrometric investigations on CBHI from twoT. reeseistrains grown under three conditions, several new discoveries were made: 1) N45 and N64 areN-glycosylated with high mannose type glycans; 2) the catalytic domain of CBHI is extensivelyO-glycosylated with hexoses andN-acetylhexosamines; 3) experimental evidence on the mannosylation of carbohydrate binding domain (other than the linker adjacent region) was found. With structural analysis, we found several glycosylation sites (such as T383, S8, and S46) are located at the openings of the substrate-binding tunnel, and potentially involve in the binding of cellulose. These investigations provide a full and comprehensive picture on the glycosylation of CBHI fromT. reesei, which benefits the engineering of CBHI by raising potential sites for modification.
