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Km And Vmax Of Alkalne Lab Report

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Km And Vmax Of Alkalne Lab Report
Jay A. Torres
With David Shbeeb and Matt Shinsky
Liberty University
1003 Misty Mountain road apt.507, V.A. 24502 jtorres99@liberty.edu words count: 1,313

Determination of Km and Vmax of Alkaline Phosphatase
Graphs will be used with the enzyme alkaline phosphatase of the unknown in the enzyme solution to Determine Km and Vmax

To Dr. Kimberly A. P. Mitchell, Editor, Liberty Journal of Cell Biology, 1971 University Blvd, Lynchburg, VA 24502.

Materials and Methods

Dilution: Alkaline Phosphatase
The objective of this laboratory is to determine the velocity of alkaline phosphatase and make a graph of rate vs substrate concentration (Michaelis Menten plot). After the wavelength was determined the para-Nitrophenyl phosphate (PNPP, Sigma Aldrich) …show more content…
After getting those results and calculating the extinction coefficient a graph was made to calculate the rate of reaction with the axes PNP (Sigma Aldrich) versus the time. The formula used to calculate the rate of reaction was v=Vmax[S]/Km+[S] (Vmax) is the maximum velocity of the enzyme, Km is the Michaelis-Menton, and [S] is the concentration of the substrate). The concentrations used in the graph are 0.5, 0.3, 0.05, 0.03, 0.005 mM. And the results for the rates of reaction were 0.4519, 0.4694, 0.2186, 0.1126, 0.0502 Mm/s.

Determining the Vmax and Michaelis-Menton constant for Alkaline Phosphatase After getting the result of the rate of reaction, the figure #3 was made with the axes of rate of reaction versus the PNPP. The reason why this graph was made is to determine the V max and the Michaelis-Menton constant using the formula K m=[S] at 1/2Vmax. The highest rate of Vmax was 0.4700 mM/s. using the formula ½ Vmax the value for the Km was determinate. After that the value of Y, which was 3.0739, was used to get the X value; and then using the equation of the hyperbola the Km was calculated. The value for the Km was 0.0500 …show more content…
The formula used to get the results was 1/v=1/Vmax+Km/Vmax*1/[S]. In order to get the Lineweaver-Burke plot, we need to get from the graph before (graph #3) the data. After solving for the Km and Vmax with the formula provided, the X and Y were solved to obtain the reciprocal values with the formula provided. The result for the y intercept was 3.0739 and for the x value was -35.5775463, and then the Vmax and Km were calculated with those results. The Vmax was 0.3252 mM/s and the Km was 0.02811 mM. As we could see the results were completely different from the Vmax and Km obtained from the Michaelis-Menton so we assume that could be on the graph that is not accurate when it tries to interpret the
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