Dear Arshad,

Mine is also same work..substrate is different..which material are used for tannase..there are different method for tannase assay. theoretically tannin will degrade to tannase and the final product is gallic acid. if your taking nmr with your sample and gallic acid then you can find out how much substrate is left over...tannase is a hexameric molecule its very difficult to get a monomer..

please do spectrum analysis in your instrument..

Thank you Christoph.

My enzyme assay method : 0.3ml substrate(concentration 0.5%)added 0.05ml enzyme and incubated 10min and added BSA solution 3ml cenrtifuged took the pellet and added 3ml of SDS-triethanol amine and 1ml of FeCl3 reagent and i messured OD at 530 nm.

enzyme activity calculation formula :

umol of substrate degraded X assay vol (4ml) / 10 min X 0.05 

My substrate tannic acid have 1701g/mol molecular weight . using this if i calculate how much umole consumed by enzyme i am getting very low value. so is any other formula is there for calculating enzyme activity

the sensitivity of measuring substrate disappearance is very low since you start from a high value to a lower value.  If you can, you should measure the appearance of product because from zero to a certain value is much more sensitive and thus accurate.

 sir,

i am also working on tannase but i got higher level of tannase production. Is this formula best suited for the same?

0.3ml substrate(concentration 0.5%)added 0.1ml enzyme and incubated 10min and added BSA solution 3ml cenrtifuged took the pellet and added 3ml of SDS-triethanol amine and 1ml of FeCl3 reagent and i messured OD at 530 nm.

enzyme activity calculation formula :

umol of substrate degraded X assay vol (4ml) / 10 min X 0.1

How can I calculate enzyme activity based on the how much.... Available from: https://www.researchgate.net/post/How_can_I_calculate_enzyme_activity_based_on_the_how_much_substrate_is_left_over_after_enzyme_reaction [accessed Aug 28, 2017].