000201662 001__ 201662
000201662 005__ 20181203023618.0
000201662 0247_ $$2doi$$a10.1007/s10570-011-9506-2
000201662 022__ $$a1572-882X
000201662 037__ $$aARTICLE
000201662 245__ $$aObserving Thermobifida fusca cellulase binding to pretreated wood particles using time-lapse confocal laser scanning microscopy
000201662 260__ $$c2011
000201662 269__ $$a2011
000201662 336__ $$aJournal Articles
000201662 520__ $$aWe report on studies of Thermobifida fusca cellulases Cel5A, Cel6B and Cel9A binding to pretreated wood particles using Confocal Laser Scanning Microscopy (CLSM). Hydro-thermal pretreated wood particles were immobilized on borosilicate substrates before fluorescently-labeled cellulase solutions at various concentrations were added. Time-lapse CLSM revealed that cellulases Cel5A, Cel6B and Cel9A quickly bound to certain areas of wood particles, slowly diffused into and adsorbed to less accessible areas, but showed little affinity for other areas of the wood. Cellulase-to-substrate association constants were estimated using a transient enzyme binding kinetics model, and were found to be in agreement with published values. In order to accurately account for the fluorescence signal of labeled enzyme mixed with wood autofluorescence, we also developed a spectral deconvolution method to separate signals from multiple fluorochromes. © 2011 Springer Science+Business Media B.V.
000201662 700__ $$aZhu, Paul
000201662 700__ $$aMoran-Mirabal, Jose M.
000201662 700__ $$0248339$$g154408$$aLuterbacher, Jeremy S.
000201662 700__ $$aWalker, Larry P.
000201662 700__ $$aCraighead, Harold G.
000201662 773__ $$j18$$tCellulose$$k3$$q749-758
000201662 909C0 $$xU12907$$0252511$$pLPDC
000201662 909CO $$pSB$$particle$$ooai:infoscience.tind.io:201662
000201662 917Z8 $$x242569
000201662 917Z8 $$x242569
000201662 917Z8 $$x249835
000201662 937__ $$aEPFL-ARTICLE-201662
000201662 973__ $$rREVIEWED$$sPUBLISHED$$aOTHER
000201662 980__ $$aARTICLE