TRYPSIN LAB
Title: The Effects of Substrate Concentration and Temperature on the Rate of Hydrolysis of the Enzyme Trypsin.Abstract: Quantitative measurements can relate both temperature and substrate concentration to the enzymatic activity of trypsin. By analyzing the data, it is suggested that at BAPNA concentrations below those corresponding to Vmax are rate limiting, as less active sights are available for adhesion. The values of Vmax and Km relate a temperate catalytic efficiency of trypsin. The temperature range of most efficiency for the enzyme was those between 36 and 54 degrees Celsius. Introduction: Enzymes are specialized proteins that aid in formation or breakdown of larger protein or multi-protein complexes. Trypsin is a pancreatic protease that digests proteins by hydrolyzing the peptide bonds in proteins. It has a high degree of specificity and will only hydrolize the peptide bonds that occur on the carboxyl side of the amino acids lysine or arginine. Generally hydrolytic reactions occur with the addition of water to breakdown a large protein into two protein fragments. Substrate concentration and temperature both would interfere and affect with the hydrolysis of Na-benzol-L-arginly-p-nitroanalide (BAPNA) into
This was the control to measure the hydrolysis of BAPNA in the absence of enzyme. Part 2: Effect of Temperature on VelocityObtain constant amounts of 10X buffer, H2O, BAPNA, and enzyme and place into cuvettes, saving the addition of enzyme until last. The results were then plotted with the absorbance being the dependent variable and the concentration the independent. The extinction coefficient was then used to convert each absorbance reading to PNA concentration. The most efficient temperature demonstrated by our experiment was that of 54C. The Lineweaver-Burke plot graph (Fig 1) estimated the value of Vmax to be 0. These two values can be determined from the double reciprocal of the Michalelis-Menton equation or the Lineweaver-Burke Plot, with the y intercept being 1/ Vmax, and the x intercept being -1/ Km. Our data showed this too is true, as product was formed at a faster rate with more enzymes, than of those solutions containing less. The slope of the linear portion of the graph represented the initial velocity of substrate hydrolysis as a function of time.
Common topics in this essay:
Manual E=A/cl,
Vmax Lineweaver-Burke,
Velocity Cuvette,
H2O BAPNA,
Na-benzol-L-arginly-p-nitroanalide BAPNA,
Introduction Enzymes,
Abstract Quantitative,
Lineweaver-Burke Plot,
Celsius Below,
Vmax Vmax,
substrate concentration,
extinction coefficient,
rate reaction,
10x buffer,
01 ml,
pna concentration,
initial velocity,
placed spectrophotometer,
ml h2o,
h2o added,
added mixed placed,
velocity substrate hydrolysis,
mixed placed spectrophotometer,
collisions enzyme substrate,
initial velocity substrate,
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Acceptance Essays
