Lineweaver–Burk plot

Study of stereospecificity in mushroom tyrosinase and the inhibiting effects of thiourea, cinnamic acid and benzoic acid BIOL/BIOC 393 L03 Dr. Judit Moldovan Submitted: Nov. 22nd, 2010 By: Jackie Minnick (Partners Amanda Verwoerd & Kersti Ojamaa) Study of stereospecificity in mushroom tyrosinase and the inhibiting effects of thiourea, cinnamic acid and benzoic acid. Jackie Minnick This paper reports experiments on the stereospecificity observed in the monophenolase and diphenolase activities

Vmax values obtained from the Michaelis Menten graphs in Figure 4 and Figure 5 do not correspond with Km and Vmax values obtained in the Lineweaver-Burk plot of LDH in Figure 3. The Vmax obtained in Figure 4 and Figure 5 were both 10.6µM/min in the absence of inhibitor whereas the Vmax in the absence of inhibitor was 0.34 µM/min in Figure 3. Lineweaver-Burk plots allow for an accurate derivation of an enzyme’s Km and Vmax through an extrapolation of reciprocal values as opposed to the Michaelis-Menten

2.8 MedCalc: MedCalc is a statistical software package designed for the biomedical sciences. It has an integrated spreadsheet for data input and can import files in several formats. 2.9 Test principle for Roche Cobas 6000: UV test Enzymatic reference method with hexokinase. Hexokinase catalyzes the phosphorylation of glucose to glucose-6-phosphate by ATP. Glucose+ATP ______HK___→ G-6-P+ ADP Glucose-6-phosphate dehydrogenase oxidizes glucose-6-phosphate in the presence of NADP to gluconate-6-phosphate

μM/mL and 60 mins, respectively. The enzyme was then reacted with varying concentrations of 4-nitrophenyl-N-acetyl-B-D-glucosamine enzyme substrate and a Michaelis-Menten plot was generated. Once it was determined that N-acetyl-B-D-hexosaminidase follows Michaelis-Menten kinetics by exhibiting a hyperbolic curve, a Lineweaver-Burk plot was generated and kinetic parameters Vmax, Km, Kcat, and Kcat/Km ratio were calculated. The parameters Vmax, Km, Kcat, and Kcat/Km ratio were determined to be 165 μm/mL/min

After obtaining all of the absorbance values and unit conversions were calculated, as shown on page 18, a rate of product formation versus time was graphed as well as a Michaelis-Menten plot. Based upon the Michealis-Menten plot, estimated values of Vmax and Km were recorded. Instead of generating a Lineweaver-Burke plot to determine the calculated values of Vmax and Km, our lab instructor provided a video for us to watch, which allowed us to determine the calculated values of Vmax and Km right on the

The Lineweaver–Burk plot is widely used to determine important terms in enzyme kinetics, such as Km and Vmax, before the wide availability of powerful computers and non-linear regression software. The plot provides a useful graphical method for analysis of the Michaelis–Menten equation: "V =" ("V" _"max" ⁡" " " [S]" )/("K" _"M" "+ [S]" ) Taking the

Muscle contraction is an essential action by most mammalian life forms performed almost all the time and requires a quick source of energy production in order to carry out this function. Muscle cells must be able to have an adequate supply of energy in the forward reaction to contract as well as an adequate supply of oxygen in the reverse reaction to replenish their cells so that energy could be continuously produced. During vigorous activity, there is a build-up of pyruvate concentration and cells

Dopachrome (oxidation of L-DOPA) (Vernier, 2015). The aforementioned inhibitors will be added to the reaction to analyze the effects of various inhibitors of Tyrosinase. Ultimately, the goal of this lab is to study enzyme kinetics as they apply to the Lineweaver-Burk delineation of

concentration of the substrate (Washington.edu). From the Michaelis Menten Kinetic equation, we have many different ways to find Km and Vmax such as Lineweaver-Burk plot, Hanes-Woolf plot, etc. However, this lab used the Lineweaver- Burk plot. By taking the reciprocal of the Michaelis Menten Kinetics equation, we can obtain the Lineweaver –Burk double reciprocal plot, which is a linear line (chemwiki.ucdavis.edu). This provides more precise way to determine Vmax and

from yellow to reddish brown. In task 2, the enzyme kinetics of yeast invertase on sucrose was studied. The absorbance values of the corresponding volumes of the solutions were measured using a spectrophotometer. Michaelis-Menten curve and Lineweaver-Burk Plot were made in order to estimate the values of Vmax and Km