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Sunday, December 13, 2009

ESTMATION OF LIPASE ACTIVITY

HOW TO ESTMATE OF LIPASE ACTIVITY


Lipase is a fat splitting enzyme. It hydrolyses fats into fatty acids and glycero

Trigycerides +Lipase enzyme------------------->Fatty Acids +Glycerin
For lipase optimum PH is around 7-7.2 and optimum 10 mp is 370c.The enzyme unit I.O amount of enzyme that liberates fatty acid which consumes 1 ml of 0.05 m alkali at 370c and optimum PH after 3 hrs. of incubation.


Requirements: - Enzyme source: 1) 2% pancreatic. 2) 5% germinated castor oil seed extract – Acetone: Alcohol 1:1, NaOH 0.05 N, substrate. (Groundnut oil oxidize)


Procedure: - To 10 ml of substrate add 10 ml of enzyme extract incubate at 37%c for 3 hrs. After incubation add 30 ml of acetone alcohol mixture to using in inactive the enzyme. Titrate fatty acid liberated using phenol ptyalin indicator. Carry out blank using inactivated blank extract.
Enzyme unit u = amount of enzyme required to produce fatly acid equivalent to 1 ml of 0.05 NaOH at end of 3 hrs.
Preparation of substrate: 100 ml oil + 400 ml H2o + 40 gm gum acacia.
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