i want introduction of LIPASE ASSAY (USING TURBIDIMETRIC METHOD)

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Expert Solution

Lipases are enzyme that can split fat by breaking the tri-glycerides into fatty acid and glycerol.

Lipase assay is a measure of the enzymatic activity of the enzyme. Here the substrate of a enzyme (like triglycerides, olive oil, triolein) is mixed in a suitable buffer in presence stabilizing agent (like 2-ethoxyethanol) to create an emulsion with a homogenous dispersion of micelles. Within the emulsion the addition of lipase begins the process of hydrolysis of fat also decreases the number of micelle present in the emulsion. Commercial grade purified lipase can be as a measure of standard lipase activity.

Higher wavelength near about 546nm can be used to measure the turbidity. The scattering of light is a measure of turbidity also the activity of the enzyme.At lower wave length the absorbance of light by the emulsion can significantly effect the the outcome of the experiment.hence other wave length can be used depends on the performed experiment.

So the main basis of the assay is the relationship between the change in turbidity and the number of micelle in the emulsion.

purity of substrate and enzyme, pH and homogeneity of the emulsion, temperature and wave length are the major factors can effect the results of lipase assay.

gladiator answered 1 week ago

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