Orlistat is a prescription drug that acts as an inhibitor fo

Orlistat is a prescription drug that acts as an inhibitor for lipase (enzyme digest lipids). Briefly design an experiment to test this statement using at least two test tubes. You should include at list of all of the materials, the step-by-step procedures, and an explanation of your predictions of the results. Conclude your experimental write-up by drawstring this question: if Orlistat is lipase inhibitor, what conclusion can you make bout digesting and absorbing fatty foods when taking this drug?

Solution

Answer:

The test of lipase inhibition by Orlistat can be performed in following way:

Materials and Reagents:

1. Mammalian lipase or substitute like porcine or bovine pancreatic lipase.

2. Substrate pNPP para-Nitrophenyl Palmitate

3. A Suitable Buffer at pH 8.0

4. Spectrophotometer

5. Cuvettes

6. Micropipettes and tips

7. Test tubes

8. Sterile water

9. Beaker, containers

Assay Protocol:

Lipase inhibition can be assayed using a pNPP substrate ( para-Nitrophenyl Palmitate) which under normal conditions (without the presence of any inhibitor) gets hydrolyzed by Lipase and the extent of hydrolysis can be monitored by change in absorbance values measured at 410nm.

Prediction of Result: If Orlistat inhibits Lipase, then in the presence of this inhibitor, the extent of pNPP hydrolysis will reduce leading to a reduction in absorbance at 410nm. A complete dose-response profile can be obtained by using different concentrations of Orlistat in the assay.

Test tubes:

1. Control tube: Contains Substrate + Standard concentration of Lipase (un-inihibited)

2. Test tube - Contains different concentrations of Orlistat drug to get a dose response curve.

3. Blank tube - Contains only Substrate and assay reagents (without enzyme) - to set the assay baseline.

If Orlistat is a lipase inhibitor, what conclusion can you make about Digesting and Absorbing fatty foods when taking this drug:

When Orlistat drug is taken it inhibits the action of Lipase as a result the triglycerides in the food are not hydrolyzed. As a result, the fat in the food is left unabsorbed and excreted as such.