Observing Thermobifida fusca cellulase binding to pretreated wood particles using time-lapse confocal laser scanning microscopy
作者： Paul Zhu , Jose M. Moran-Mirabal , Jeremy S. Luterbacher , Larry P. Walker , Harold G. Craighead
DOI: 10.1007/S10570-011-9506-2
关键词:
摘要: We report on studies of Thermobifida fusca cellulases Cel5A, Cel6B and Cel9A binding to pretreated wood particles using Confocal Laser Scanning Microscopy (CLSM). Hydro-thermal were immobilized borosilicate substrates before fluorescently-labeled cellulase solutions at various concentrations added. Time-lapse CLSM revealed that quickly bound certain areas particles, slowly diffused into adsorbed less accessible areas, but showed little affinity for other the wood. Cellulase-to-substrate association constants estimated a transient enzyme kinetics model, found be in agreement with published values. In order accurately account fluorescence signal labeled mixed autofluorescence, we also developed spectral deconvolution method separate signals from multiple fluorochromes.
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