A rapid microassay to evaluate enzymatic hydrolysis of lignocellulosic substrates.
Current attempts to produce ethanol from lignocellulosic biomass are focused on the optimization of pretreatment to reduce substrate recalcitrance and the improvement of enzymes for hydrolysis of the cellulose and hemicellulose components to produce fermentable sugars. Research aimed at optimizing both aspects of the bioconversion process involves assessment of the effects of multiple variables on enzyme efficiency, resulting in large factorial experiments with intensive assay requirements. A rapid assay for lignocellulose hydrolysis has been developed to address this need. Pretreated lignocellulose is formed into handsheets, which are then used to prepare small disks that are easily dispensed into microtiter plates. The hydrolysis of cellulose to glucose is estimated using an enzyme-coupled spectrophotometric assay. Using disks prepared from ethanol organosolv pretreated yellow poplar, it is shown that the assay generates data comparable with those produced by hydrolysis of pretreated yellow poplar pulp in Erlenmeyer flasks, followed by HPLC analysis of glucose. The assay shows considerable time and cost benefits over the standard assay protocol and is applicable to a broad range of lignocellulosic substrates.
Berlin
Alex
A
Department of Wood Science, Biotechnology and Biomaterials Group, Faculty of Forestry, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4. alex.berlin@ubc.ca
Maximenko
Vera
V
Bura
Renata
R
Kang
Kyu-Young
KY
Gilkes
Neil
N
Saddler
Jack
J
D002384:Catalysis; D044602:Cellulases / Q000737:chemistry / Q000378:metabolism*; D002482:Cellulose / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D000431:Ethanol / Q000737:chemistry; D005947:Glucose / Q000032:analysis / Q000378:metabolism; D006868:Hydrolysis; D008031:Lignin / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D031567:Liriodendron / Q000737:chemistry; D008832:Microchemistry / Q000379:methods*; D009005:Monosaccharides / Q000032:analysis; D010209:Paper; D010407:Penicillium / Q000201:enzymology; D013379:Substrate Specificity; D014242:Trichoderma / Q000201:enzymology; D014934:Wood
D009005:Monosaccharides; C036909:lignocellulose; D000431:Ethanol; D002482:Cellulose; D008031:Lignin; D044602:Cellulases; D005947:Glucose