期刊
BIOTECHNOLOGY AND BIOENGINEERING
卷 110, 期 1, 页码 47-56出版社
WILEY
DOI: 10.1002/bit.24604
关键词
cellulase; Thermobifida fusca; surface diffusion; single molecule; fluorescence recovery after photobleaching; microscopy
资金
- USDOE [GO18084]
- New York State Empire Development Corporation
Elucidation of cellulasecellulose interactions is key to modeling biomass deconstruction and in understanding the processes that lead to cellulase inactivation. Here, fluorescence recovery after photobleaching and single molecule tracking (SMT) experiments are used to assess the surface diffusion of Thermobifida fusca cellulases on bacterial micro-crystalline cellulose. Our results show that cellulases exhibit limited surface diffusion when bound to crystalline cellulose and that a large fraction of the cellulases remain immobile even at temperatures optimal for catalysis. A comparison of our experimental results to Monte Carlo (MC) simulations, which use published diffusion coefficients to model cellulase displacements, shows that even those enzymes that are mobile on the cellulose surface exhibit significantly slower diffusive motions than previously reported. In addition, it is observed that the enzymes that show significant displacements exhibit complex, non-steady surface motions, which suggest that cellulosebound cellulases exist in molecular states with different diffusive characteristics. These results challenge the notion that cellulases can freely diffuse over cellulose surfaces without catalyzing bond cleavage. Biotechnol. Bioeng. 2013; 110: 4756. (C) 2012 Wiley Periodicals, Inc.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据