Molecular Details from Computational Reaction Dynamics for the Cellobiohydrolase I Glycosylation Reaction
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- 作者：Christopher B. Barnett ; Karl A. Wilkinson ; Kevin J. Naidoo
- 刊名：Journal of the American Chemical Society
- 出版年：2011
- 出版时间：December 7, 2011
- 年：2011
- 卷：133
- 期：48
- 页码：19474-19482
- 全文大小：1119K
- 年卷期：v.133,no.48(December 7, 2011)
- ISSN：1520-5126
文摘
Glycosylation of cellobiose hydrolase I (CBHI), is a key step in the processing and degradation of cellulose. Here the pathways and barriers of the reaction are explored using the free energy from adaptive reaction coordinate forces (FEARCF) reaction dynamics method coupled with SCC-DFTB/MM. In many respects CBHI follows the expected general GH7 family mechanism that involves the Glu-X-Asp-X-X-Glu motif. However, critical electronic and conformational details, previously not known, were discovered through our computations. The central feature that ensures the success of the glycosylation reaction are the Glu212 nucleophile鈥檚 hydrogen bond to the hydroxyl on C2, of the glucose in the 鈭? position of the cellulosic strand. This Glu212 function restricts the C2 hydroxyl in such a way as to favor the formation of the 4E ring pucker of the 鈭? position glucose. A frontier molecular orbital analysis of the structures along the reaction surface proves the existence of an oxocarbenium ion, which has both transition state and intermediate character. The transition state structure is able to descend down the glycosylation pathway through the critical combination of Asp214 (HOMO), ring oxygen (LUMO), and Glu212 (HOMO), anomeric carbon (LUMO) interactions. Using the fully converged FEARCF SCC-DFTB/MM reaction surface, we find a barrier of 17.48 kcal/mol separating bound cellulose chain from the glycosylated CBHI. Taking recrossing into account gives kcat = 0.415 s鈥? for cellobiohydrolase glycosylation.