Problem. Determine KM and Vmax for an enzyme from the following data : S(mM) vo(HM.s²) 1/S (mM) 1/v.(µM.s*) 1 2.5 1.00 0.40 2 4 0.50 0.25 5 6.3 0.20 0.16 10 7.6 0.10 0.13 20 9 0.05 0.11

Please explain your reasoning for every answer

a. What is the Vmax of this enzyme WITHOUT inhibitor? Please show your work.   b. What is the Km of this enzyme WITHOUT inhibitor? Please show your work. c. The specificity constant of enzyme X is 8 x 10^7 /(M * seconds) What is the kcat of enzyme X WITHOUT inhibitor? Please show your work d. What was the concentration of enzyme used for measuring the kinetics of enzyme X WITHOUT inhibitor? Please show your work

Answer #2

Part 1: Assess the following partial results section below by editing it for brevity by omitting any unnecessary parts (1 point), explain why you decided to remove certain sections (1 point): To evaluate inhibitory effects of the selected molecules, 10mM stock solutions of each molecule were prepared in DMSO. A reaction mixture (200μl) was prepared with the same formula optimized for the enzyme activity assay (0.1 M Tris-HCl ph 8, 0.1 M KCI, 25 mM NaCl, 0.25 mM ATP, and two units of inorganic yeast pyrophosphatase) with 10 µM of the sample molecule. The reaction mixture was incubated for 20 minutes at ambient temperature. Enzymatic reaction was triggered by addition of the substrate B (0.2 mM) and the absorbance of the product was monitored at 290 nm for 10 minutes. Six out of 15 sample molecules showed appreciable inhibition at 10 μM (Figure 5). Three of the molecules, A3, A6, and A7 exhibited more than 50% inhibition of the enzyme activity and were further diluted to find the minimal…

By what factor is sample A2 more concentrated than sample A1?

Michaelis Menton Plot and a Lineweaver-Burke plot using this data