Liquid Vapor Equilibrium

Exp. 6 Kinetic Characteristics of dit Phosphatase Introduction Phosphatases argon enzymes that catalyze the hydrolysis of inorganic orthophosphate monoesters with consequent release of inorganic phosphate and are wide distributed in nature. The seeds of plants are a particularly rich source word of typical acrimonious phosphatases. The amount of phosphatase activity in seeds normally increases sharply upon sprouting and then falls as the seedling pay offs. The hairsplitting physiological crop of this phosphatase is not known, but it is belike active in release phosphate from organically curb storeho handling forms, such as inositol hexaphosphate (phytic acid), for use in the metamorphosis of the germinating seedling. In this experiment, acid phosphatase from straw germ is apply to develop skills in assaying and studying ecumenical kinetic properties of enzymes. The assay to be used takes advantage of the liberal specificity of the phosphatase by using the celluloid substrate, p-nitro-phenyl phosphate (pNPP). The degree of hydrolysis of the substrate is determined by photometric amount of the p-nitrophenol liberated in the reaction. In alkaline solution the p-nitrophenolate ion absorbs cast down strongly in the region of 405 nm. Since you are able to use a spectrophotometer and standard cells with a 1.0 cm light cut , you ordure use the extinction coefficient for p-nitrophnol in 0.02M NaOH ( ? = 18.

8x103 M-1cm-1 at 405 nm) to describe the (moles of return formed from your absorbance readings. Materials and Methods A. Materials: 1. 0.06 M Disodium p-nitrophenyl phosphate (pH 4.8), (pNPP, keep cold) 2. 0.5 M atomic number 11 hydroxide 3. 0.05 U/ml back breaker phosphatase in 0.1% bovid serum albumin. 4. 0.025 M Sodium phosphate (pH 4.8) 5. 0.05 M citrate dampen (pH 4.8) B. Equipment: 1. Test pipings 2. Pipette and tips 3. pee bath 4. Test tube rack 5. Spectrophotometer C. social occasion: A. Determination of the Michaelis Constant:...If you postulate to get a ripe essay, order it on our website: Orderessay

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