Review of lignocellulolytic enzyme activity analyses and scale-down to microplate-based assays

• Existing enzyme activity assays were reviewed for four lignocellulolytic enzymes.
• Activity calculation methods were defined for each enzyme.
• 96-well plate adapted methods were identified based on a statistical approach.

With the increasing use of enzymes in environmental applications, there is a need for analytical methods adapted to large factorial experiments. Existing reference methods are chemical and labor intensive and unsuitable to analyze in parallel a large number of samples. Based on an extensive literature review and on experimental results, this work compares reference and microplate adapted methods to define the most adequate filter paper, carboxymethylcellulase, β-glucosidase and xylanase activity tests. In the adapted methods, the total reaction volume was reduced from 2.2–24.5 mL to 0.21–0.24 mL. Statistical analysis of the activities measured on enzyme mixtures by applying the 96-well plate reduced methods showed that they were not significantly different to the activities obtained with reference tests.

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