Effect of temperature on Enzyme Activity

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Chemical and Biochemical Engineering
Chemical and Biochemical Engineering 2290A/B
Mita Ray

WEEK 3 Aim: To study the effect of temperature on enzyme reaction rate Protocol: End point Assay method Assay mixture: 3.9ml Tris HCl pH 9.0 1.0ml substrate solution At T=0 , add 0.1ml alkaline phosphatase (enzyme) At T=4min, add 2ml 2N NaOH Read absorbance at end of 4min at 410nm. Repeat 3 times at each temperature Observation (End point assay method) Sr. No TEMPERATURE (˚C) Observation (Absorbance @ 410nm) t= 4 min 1 2 3 1 0 2 25 3 35 4 45 5 55 6 65 Calculate: a. For each treatment, plot product concentration vs. time. b. For each treatment, determine reaction rate using the graphical method and put the appropriat
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