In situ, rapid, and temporally resolved measurements of cellulase adsorption onto lignocellulosic substrates by UV-vis spectrophotometry.
This study demonstrated two in situ UV-vis spectrophotometric methods for rapid and temporally resolved measurements of cellulase adsorption onto cellulosic and lignocellulosic substrates during enzymatic hydrolysis. The cellulase protein absorption peak at 280 nm was used for quantification. The spectral interferences from light scattering by small fibers (fines) and particulates and from absorptions by lignin leached from lignocelluloses were corrected using a dual-wavelength technique. Wavelengths of 500 and 255 nm were used as secondary wavelengths for correcting spectral interferences from light scattering and absorption of leached lignin. Spectral interferences can also be eliminated by taking the second derivative of the measured spectra of enzymatic hydrolysate of cellulose or lignocelluloses. The in situ measured cellulase adsorptions in cellulose and lignocellulose suspensions by these two spectrophotometric methods showed general agreement with batch sampling assayed by the Bradford method. The in situ methods not only eliminated tedious batch sampling but also can resolve the kinetics of the initial adsorption process. The measured time-dependent cellulase adsorptions were found to follow pseudo-second-order kinetics.
State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou, China.
Hao
H
Liu
J Y
JY
Zhu
X S
XS
Chai
D000327:Adsorption; D002138:Calibration; D002480:Cellulase / Q000737:chemistry* / Q000378:metabolism*; D002482:Cellulose / Q000737:chemistry* / Q000378:metabolism*; D006868:Hydrolysis; D008031:Lignin / Q000737:chemistry* / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D013535:Suspensions; D013997:Time Factors
D013535:Suspensions; C036909:lignocellulose; D002482:Cellulose; D008031:Lignin; D002480:Cellulase