1. What are discrepancies between calf intestinal mucosa alkaline phosphatase and human alkaline phosphatase? 2. A...

1. What are discrepancies between calf intestinal mucosa alkaline phosphatase and human alkaline phosphatase?

2. A solution was made up with p-Nitrophenyl phosphate (pNPP), 0.1 M borate buffer at pH 9.5 and and alkaline phosphatase. This solution has good enzyme activity. Why does changing the pH to 9 decrease the apparent activity of alkaline phosphatase?

3. solution was made up with p-Nitrophenyl phosphate (pNPP), 0.1 M borate buffer at pH 9.5 and and alkaline phosphatase. 50 uM of phosphate was added into the solution. Does the addition of phosphate to the assay mix cause competitive or non-competitive inhibition? Is this expected?

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gladiator answered 1 year ago

Experiments A and B are 2 experiments performed by mixing alkaline phosphatase enzyme with Pnpp substrate....

Experiments A and B are 2 experiments performed by mixing alkaline phosphatase enzyme with Pnpp substrate. The reaction was run for 5 minutes for each tube, then stopped by adding NaOH. A)Changing enzyme concentration Tube Reaction Buffer (mL) 5.4 mM of pNPP substrate (mL) 0.002 mg/ml of AP enzyme (mL) 3M of NaOH (mL) Absorbance Final Enzyme concentration (mM) Velocity (umol/min/mL) 1 3.41 0.04 0.05 0.875 0.284 2 3.36 0.04 0.1 0.875 0.387 3 3.31 0.04 0.15 0.875 0.509 4...

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