Hello
I need help with some enzymatic assays.
I performed the methodologies to evaluate the activity of chitinase, B-1,3-glucanase, Protease and NGAse from Trichoderma harzianum. I performed the assays in microplates and read the absorbance in spectrophotometer according to some publications. But now, unfortunatelly, I do not know how to work with the results. How can I calculate enzymatic activity? What parameter do I need to obtain the results?
I could not find anything about the calculation in literature.
Can anyone help me?
Thank you very much!
This should be in the paper describing the assay. Basically, you need the molar extiction coefficient of your coloured product at the wavelength you use to measure. Then the concentration is calculated from Lambert-Beer's law.
The book Biochemical Calculations by I.H.Segel has helped generations of newbees with problems like this.
For chitinase:use N-acetylglucosanine (5mg/ml) for calibration curve, then from the reaction time(t1-t0) you will have a value which you have to incorporate into your calibration curve.
If you need other information specifically say it.
Good luck
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