1- (a) How could you design a colorimetric method to measure alpha-amylase activ
ID: 309770 • Letter: 1
Question
1- (a) How could you design a colorimetric method to measure alpha-amylase activity? (b) What would you use as a control? (c) At what wavelength should the amylase activity you would observe using your method?2- You want to know whether alpha-amylase is more active in the stomach (pH 2) or the intestines (pH 7). (a) How would you set up an experiment to test this? 1- (a) How could you design a colorimetric method to measure alpha-amylase activity? (b) What would you use as a control? (c) At what wavelength should the amylase activity you would observe using your method?
2- You want to know whether alpha-amylase is more active in the stomach (pH 2) or the intestines (pH 7). (a) How would you set up an experiment to test this? (a) How could you design a colorimetric method to measure alpha-amylase activity? (b) What would you use as a control? (c) At what wavelength should the amylase activity you would observe using your method?
2- You want to know whether alpha-amylase is more active in the stomach (pH 2) or the intestines (pH 7). (a) How would you set up an experiment to test this? (b) What would you use as a control? (c) At what wavelength should the amylase activity you would observe using your method?
2- You want to know whether alpha-amylase is more active in the stomach (pH 2) or the intestines (pH 7). (a) How would you set up an experiment to test this?
Explanation / Answer
1.a)
COLORIMETRIC METHOD TO MEASURE ALPHA AMYLASE ACTIVITY
Starch is the polysachharide which consists of repeated units of glucose as amylose and amylopectin units, with the action of alpha amylase activity, the glycosidic bonds is cleaved and produces the maltose,maltotriose and glucose as sub units.
The present experiment is to measure the alpha amylase activity, so if we take the enzyme concentration as constant and standard concentrations of substrates i.e.,glucose and test sample starch, so the amylase activity is measured which exploited in the standard graph, to know the alpha amylase activity. The optical density values are measured using coloriemeter which measure at 585nm intensity.
b & c ) Glucose is control. When the starch is hydrolysed which invovles two step process,
step 1 :- First starch is converted to glucose by alpha glucosidase and hydrogen peroxide by glucose oxidase
step 2 : The hydrogen peroxide concentration is measured colorimetrically at 585 nm.
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