i use the enzyme with ≥20,000 U/g on mill waste water for the first time.
how i determine the amount of enzyme to be used ? or how to determine the percentage of enzyme to add? and what puffer i can use to make enzyme stock
These are all good questions but to try an answer we need more information. For starter, what enzyme? From what company?
i want to use tow enzyme
1. Cellulase that break down the cellulose molecule into monosaccharides.
2. Horseradish peroxidase to break down Glycoside bond between phenol and sugar.
the enzyme from sigma
Dear Sabreen
You can find the answer related cellulase in this article at the mathodololgy section and subsections of FPA and enzymatic hydrolysis.
https://ojs.cnr.ncsu.edu/index.php/BioRes/article/view/BioRes_11_3_5687_Ethaib_Microwave_Dilute_Acid_Pretreatment_Enzymatic_Hydrolysis
For more details keep in touch
The problem in such cases is that the waste water contains x mM of an offending compound, which have to be brought down to y mM. The amount of enzyme required depends on the volume of waste water, the concentration difference x-y and the incubation time allowed.
1 U of enzyme breaks down 1 µmol of substrate per min under substrate saturation (this is an oldfashioned way to express activity, nowadays we use katal = mol/s). As the reaction proceeds, [S] and hence v decrease, if you plot [S] over time, you get a hyperbola. Eventually, you'll reach the equilibrium concentration of S and P, then the overall reaction velocity becomes 0 and [S] no longer changes.
I'd determine these hyperbolae for different [E] and then select the lowest [E] that gives the required degree of purification in the time allowed. Note that under the conditions of waste water treatment, you may not reach the enzymatic activity stated for the enzyme, because of temperature, pH, substrate concentration, presence of inhibitors... So there is no way to predict the optimal [E], you need to do the experiments.
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