Enzyme Kinetics

Enzymes are catalytic proteins that are essential in beneficent metabolism, as they help facilitate many of the biochemical re performions occurring in the body by lowering the activation energy that is incumbent for each reaction to occur. Various factors, such as temperature and pH, defy to the enzyme activity and can be directly regulated to act the overall rate of reaction. The purpose of this experiment is to determine the best physical conditions of the glyocolytic enzyme brawniness imbibe dehydrogenase (mLDH), which aids in the decrease and renascence of pyruvate to hold and the oxidation of NADH in skeletal tendon, and to give come forth the effects of altering temperature and pH on the activity of the enzyme mLDH. Methods In couch to complete this experiment and determine the optimum pH and temperature of the enzyme mLDH, 16.2 mM pyruvate in dH2O was place in 0.2 mM NADH in 54 mM phosphate buffer solutions of varying temperatures and pH levels, in acco rdance with the following reaction: pyruvate + NADH + H+ lactate + NAD+ In this experiment, 900 µL of NADH solution with a pH 5.5 (at about 25°C) and 25 µL of 1:2000 dilution of muscle homogenate (containing mLDH) were mixed inside a cuvette. After waiting a mo, 50 µL of pyruvate was then added to the cuvette and then mixed.

The cuvette was then fixed in a spectrophotometer and the initial absorbance of the solution was heedful. Subsequent absorbance measurements were taken each splendid over a three minute period. The overall enzyme activity was thrifty indirectly through the absorbance of the solut ion. Absorbance was measured using a spectro! photometer, an instrument that measures the light absorption of a solution at a given wavelength with a spectrophotoelectric cell. in the first place each absorbance measurement was taken, the spectrophotometer was zeroed at 340 nm by a cuvette filled with deionized water to create a basis of par and to neglect the absorbance of the water. NADH absorbs the...If you want to get a full essay, order it on our website: BestEssayCheap.com

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