Suggested improvements to the standard filter paper assay used to measure cellulase activity.
Two suggestions can be found in the literature to improve the reproducibility of the Mandels' filter paper assay: add supplemental cellobiase and increase the boiling time for color development. Here we provide data that strongly supports adding supplemental cellobiase. Adding supplemental cellobiase increased assay response by 56%. Cellulases from different sources have different cellobiase activities, which would cause significant variation in the assay response. There is no need for additional boiling time-5 minutes is sufficient. For maximum reproducibility, it is essential that the water bath vigorously boil so that temperature excursions are minimized.
Coward-Kelly
Guillermo
G
Department of Chemical Engineering, Texas A & M University, College Station, Texas 77843-3122, USA.
Aiello-Mazzari
Cateryna
C
Kim
Sehoon
S
Granda
Cesar
C
Holtzapple
Mark
M
D015374:Biosensing Techniques / Q000379:methods*; D002480:Cellulase / Q000032:analysis* / Q000737:chemistry*; D003124:Colorimetry / Q000379:methods*; D004789:Enzyme Activation; D005374:Filtration / Q000379:methods*; D006358:Hot Temperature; D011786:Quality Control; D012459:Salicylates / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013379:Substrate Specificity; D001617:beta-Glucosidase / Q000737:chemistry*
D012459:Salicylates; C027011:3,5-dinitrosalicylic acid; D001617:beta-Glucosidase; D002480:Cellulase