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alpha amylase - DNS assay

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#1 Microhead



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Posted 29 January 2011 - 11:26 AM

I have determined a glucose concentration of 0.1mg/ml of glucose from the DNS standard curve for 50 ml overnight Bacillus culture. This is equivalent (from my understanding) to 0.1 enzyme units (an enzyme unit is defined as the amount of enzyme releasing 1 mg of glucose equivalent from the substrate per 30 min at 40C). I estimated 0.1 x 1000ml/50ml (over night culture) = 100 U/L of alpha amylase (Bacillus culture)???

0.1 mg glucose = 100ug of glucose therefore 100ug glucose /180 (mol wgt of glucose) X 1/30 (1 min divide by 30 min) = 0.019 international units or approx. 0.02 U

Are the calculations correct?
Bogged down in which is the best way to present the alpha amlyase concentration or amount??
Is there a way to back calucate from U/L to I.U to check calculations?

Help appreciated.

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