How do you use Lineweaver Burk plot to determine the Vmax and Km for both the uninhibited enzyme and the enzyme with “Inhibitor A”? Table 4.3Mean Rate (mm/minute) of ONPG hydrolysis catalyzed byß-Galactosidase at various ONPG concentrations with andwithout inhibitor. Each mean is the average of 36 replicates.ONPGConcentration(MM)0357.51015203040Mean Reaction Rate (mm/minute)SEMNoInhibitor (No Inhibitor)0.0000.0000.0070.0000.0120.0010.0150.0010.0200.0010.0270.0010.0330.0010.0450.0020.0460.001InhibitorA0.0000.0040.0060.0090.0120.0170.0210.0320.034SEM(Inhibitor A)0.0000.0000.0000.0010.0010.0010.0010.0010.001 1/Mean Reaction Rate (1/mm/min)300250200150100500-0.050.00◆no inhibitorInhibitor A0.050.100.150.201/[ONPG] (1/mM)y = 733.66x + 10.9190.25y = 390.8x + 10.770.300.35Figure 4.4: Lineweaver-Burk plot showing the effect of an inhibitor on hydrolysisof ONPG by b-galactosidase. Data points are the means of 36 replicates.0.40

The Lineweaver-Burk plot, also known as the double reciprocal plot, is a graphical representation of…

Answered: Lineweaver Burk plot to determine the…

Science

Biochemistry

Lineweaver Burk plot to determine the Vmax and Km

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

See similar textbooks

Similar questions

For the following problem, indicate whether enzyme is repressed (-) or produced (+). Please show all possible outcomes.
Table 1. Kinetic parameters of a chosen model wild-type (WT) enzyme and three catalytically-improved mutants (M1 to M3). Enzyme variants k (8¹) WT MI M2 M3 50 500 1000 50 K₁, (μM) 10 100 100 1 k/K (HM¹¹ s¹) 5 The study above was done on a wildtype enzyme and three genetically-engineered mutants. The purpose was to find a mutation to improve the catalytic efficiency of the enzyme for industrial use. Which is the most accurate statement concerning the data? M2 is the optimal choice when substrate concentrations are saturating. The wildtype enzyme is still preferable to M3 when S<
why is non-competitive inhibiton a special type of mixed inhibiton?
(b) Both laboratories used 10 micrograms of protein each in their kinetic assays. Protein concentrations weredetermined by the Bradford protein assay. Assay conditions employed in the two labs (pH, temperature,etc.) were also identical. What would be the most plausible cause for the discrepancy in the Vmax valuesfor the compound I? Explain.Recall that the Bradford assay measures total protein amounts in sample solution based on complexformation between a dye and proteins. Also, the assay solution used in both labs does not contain anyinhibitors.
What is an electrochemical gradient? What purpose does it serve?
calculate the volume of stock solutions required to make up the buffer solutions that will be used for protein purification. The solutions you need to prepare for purification are: i. Binding Solution A: make up 50 mL 50 mM HEPES buffer (pH 7.5), 300 mM NaCl, 5mM imidazole, 5% (v/v) glycerol ii. Wash Solution B: make up 50 mL 50 mM HEPES buffer (pH 7.5), 300 mM NaCl, 75mM imidazole, 5% (v/v) glycerol iii. Elution Solution C: make up 10 mL 50 mM HEPES buffer (pH 7.5), 300 mM NaCl, 500 mM imidazole, 5% (v/v) glycerol please show your working . Thnk you
The natural enzyme is recreated by mixing isolated regulatory and catalytic subunits of ATCase. What biological relevance does this observation have?
Two-dimensional gel electrophoresis of proteins in a cell extract provides a qualitative way to compare proteins with respect to intra- cellular abundance. Describe a quantitative approach to the deter- mination of number of molecules of an enzyme per cell.
Describe reaction between AP and PNPP and how is used to determine the kinetic parameters of AP. i. İndicate the value for the activity of AP(which is 3.08) What does it mean? Is your enzyme active or not? What does it say about the purity of your enzyme? ii. How the purification process affects the activity?
In a pUC19 digest for 1 ug of pUC19 (DNA conc. 282ng/ul) using 10X Cutsmart buffer, pure water and BamHI enzyme in a total volume of 40 ul, how much of each solution is added to the total volume?
You are working at a major pharmaceutical company, and you have purified an engineered enzyme, and you need to determine the protein stability (kJ/mol). Accordingly, you have collected equilibrium unfolding data using the chemical denaturant urea at 20°C, pH 7.5 and I atmosphere using fluorescence spectroscopy. a) Determine the protein stability (AG [kJ/mol]) via nonlinear least squares (using the available data). b) Write the partial differential equation (i.e., general functional form, variable, constants). c) Describe what would happen to the isotherm if the temperature was increased to 25 °C. Urea [M] 0 0.25 0.5 0.75 1 1.25 1.5 2 2.25 2.5 2.75 3 3.25 3.5 3.75 4 4.25 4.5 4.75 5 FL 340nm 316 315.5 315 314.5 314 315 300 275 225 125 75 60 55 54 53 52.5 52.25 52 51.75 51.5 dG = Vdp-SdT+μA dnд + μB dng dGp = μA dnA+ μg dng A = Folded State B = Unfolded State
An enzyme with a Km of 3X10-5 M has a lower affinity for its substrate than an enzyme with a Km of 2x10-6 M. Is this true or false?