pectinase assay - pectinase acitvity (Jun/05/2010 )

can anyone help me to know how to determine the pectinase acitivity
iam working on pectinases

i found a problem in assaying pectinase. Iam using Dinitrosalicylic acid method for estimation of reducing sugars

can anyone help me

-kesavaraj-

what is the problem that you encountered?

here is an assay from worthington: pectinase assay.

-mdfenko-

respected sir

thanks for ur reply. i feel difficult in calculating the activity. can u plz provide me the source for calculating the activity of enzymes.

thanking you

-kesavaraj-

in order to help you calculate we need to know the format of the data, reaction specifics (such as enzyme concentration, substrate concentration, etc), etc, and what units in which you want the results.

-mdfenko-

iam using an enzyme concentration of 10mg/ml and substrate concentration of 5mg/ml

ultimately when it is read in a spectrophotometer it gives a value for the enzyme and standards D-galacturonic acid.

now how to calculate the amount of D- galacturonic acid liberated expressed in micromoles.

plz suggest me some books or material regarding this calculation. it will be so helpful for me

i also tried with DNSA method to estimate the reducing sugars, here also some problem in calculation.
i dont have much basics in biochem so please help me.

thanking you

-kesavaraj-

are those stock concentrations or concentration in the assay?

product (looking for increase) or substrate (looking for decrease) standard curve (give specific information)?

you also need the formula weight.

-mdfenko-