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Enzyme Activity Assay: Microplate Reader Method for Soil Analysis

A simplified method to assess enzyme activity in soil samples using a fluorometric substrate and a microplate reader. Soil samples are processed in a series of steps to measure enzyme activities.

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Enzyme Activity Assay: Microplate Reader Method for Soil Analysis

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  1. SSI Enzyme Activity Assay

  2. Method in a nutshell (to microplate reader) Fluorescence Substrate capable of fluorescence When enzyme binds substrate we have fluorescence Enzyme (of course!!)

  3. Mix soil and buffer in a blender • Transfer soil solution to a 96-well plate • Add substrates into the wells • Incubate at 25oC for 80 min • Centrifuge the plate to seperate soil and supernatant • Transfer the supernatant to another plate • Read fluorescence in a plate reader S2 S3 S4 S5 S6 S7 S8 S1 B-glucosidase substrate N-acetyl-glucosaminidase substrate Phosphatase substrate Cellulosebreakdown Chitinbreakdown Phosphatebreakdown Makeyourguess…

  4. B-glucosidase activity higher in SGS litter quality …. Pulse? N-acetylglucosaminidase higher in Forest fungal pathway enhancement

  5. Phosphatase higher activity no idea!

  6. Anattempt of conclusion…… Forest: more microbes (data fromothergroup) butlower LICOR measurements and lowerenzymeactivity Zzzzzzz

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