Enzyme Kinetics of Beta-Galactosidase

From this graph and chart we can see that the higher the concentration the higher the absorbance, all the different concentrations were tested at the same wavelength (625nm). Also we can determine our unknown substances concentration by using the absorbance we got for it. The red dot on the graph followed by the line towards the horizontal axis indicates that the concentration of fast green was 34% or 5.1x10-3.

The same solution of 0.5 ml BSA was then added from test tube 1 to the test tube 2 after being properly mixed, and from test tube 2 the solution was being added to test tube 3, and so forth all the way up to test tube 5, with the same exact procedure. From the last tube, we then disposed the 0.5 ml solution. After above procedures, we now labeled another test tube “blank”; 0.5 ml blank distilled water was purred into the tube with the serial dilution of 1:10. We also had a tube C labeled “unknown” with the same 0.5 ml of solution. And after adding 5ml of Coomassie Blue to each tube (1-5) and to the blank, the result of absorbance was read at 595 nm.

Question: How does changing enzyme concentration or temperature affect the reaction time of enzyme activity?

Make sure to use the same type of cuvette to keep the width consistent and to prevent any experimental error from arising. Obtain 5 of the same type of cuvettes and pre-rinse them thoroughly. Label them numbers one through five in increasing molarity. Then, fill each of the cuvettes with one of the five solutions you created back in Part A. We will first examine the solution that exhibits the highest concentration or molarity. Make sure to wipe the outside of the cuvette with a Kimwipe before placing into the SpectroVis Plus device. Observe the graph that is generated and make sure to take note where the maximum absorbance takes place.

The dark, navy blue colored graph represented the absorbance curve for the S1 sample. The red colored graph represented the absorbance curve for the S2 sample. The green colored graph represented the absorbance curve for the P1 sample. The purple colored graph represented the absorbance curve for the P2 sample. The gaps between the P2 curve was due to the oversaturation that led to the inconclusive spectrophotometer readings. The blue colored graph represented the absorbance curve for the P1 low salt sample. The orange colored graph represented the absorbance curve for the P2 low salt sample. The light blue colored graph represented the absorbance curve for the P1 medium salt sample. The light pink colored graph represented the absorbance curve for the P2 medium salt sample. The light green colored graph represented the absorbance curve for the P1 high salt sample. The light purple colored graph represented the absorbance curve for the P2 high salt

Spectrophotometers are quite precise instruments allowing for five significant figures to be obtained. The major source of error in this lab is how small the concentration of the reagents are.

The purpose of this experiment was to record catalase enzyme activity with different temperatures and substrate concentrations. It was hypothesized that, until all active sites were bound, as the substrate concentration increased, the reaction rate would increase. The first experiment consisted of five different substrate concentrations, 0.8%, 0.4%, 0.2%, 0.1%, and 0% H2O2. The second experiment was completed using 0.8% substrate concentration and four different temperatures of enzymes ranging from cold to boiled. It was hypothesized that as the temperature increased, the reaction rate would increase. This would occur until the enzyme was denatured. The results from the two experiments show that the more substrate concentration,

Abstract: Enzymes catalyze reactions, by lowering the activation energy of the transition state. This allows the substrate to get to the transition state more quickly than without the enzyme, thereby forming product more often. The experiment begins with questions about environmental changes that an enzyme may encounter, in particular what would these environmental factors do to the enzyme β-Galactosidase, which was the enzyme tested. The effectiveness of β-gal was tested against substrate specificity, concentration of substrate, temperature, and pH. When tested for substrate specificity, one tube contained sucrose the other OPNG, all tubes were incubated at about 37ºC for 90 seconds. The enzyme was tested in different temperatures, with a measurement of the reaction time.

Incorporation of assay controls included setting up a spectrophotomer and running the chart recorder with a full-scale deflection before the start of the assay. The set recorder had a corresponding value of 1 for the change in the absorbance. Therefore, prior testing was done to observe whether a change occurred in the readings. This helped to indicate that the results were valid, as they could have been affected by a fault during the setting up of the spectrophotometer. On the other hand this was considered as one of the controls for the experiment. Nevertheless, a new cuvette had to be used for each assay.

It was the purpose of this experiment to successfully extract the enzyme, -galactosidase from the mutated strain of E. coli sample via a multi-step protein purification process that targeted the enzymes properties of solubility, size and differences in charge. Additionally, the goal of purification was to ensure maximum enzyme production and purity at minimal loss to the enzymatic

There are many factors that may affect enzyme reaction rates. The pH and temperature of the environment that the enzyme is in may denature the enzyme if it is not in the optimal area for the specific enzyme. If the pH and temperature of the environment is in the optimal area than the enzyme will work at its best if not than reaction rates may slow down. In competitive inhibition a competitive inhibitor mimics the substrate. This causes the substrate not to enter the active site of the enzyme. Likewise there is noncompetitive inhibition where a

The purpose of this experiment was to test the effects that temperature, pH, and substrate

concentration, record the absorbance readings at a fixed wavelength, and plot the absorbance vs. concentration data. The wavelength of 520 nm was selected for experiment Part

To understand this week’s experiment one must first understand what a spectroscope is and what it does. With this understanding in hand, one would gain a deeper appreciation for this lab and its intended lesson. “A spectroscope is a device that measures the spectrum of light”

“Spectrophotometry is a method to measure how much a chemical substance absorbs light by measuring the intensity of light as a beam of light passes through sample solution” ( ChemWiki). Many chemists and biologists use the principles of spectrophotometry in everyday experiments to provide results and insight into what they are presently studying. Created in 1941, the very first spectrophotometer was called the Beckman DU UV-Vis, and was totally affordable. Despite being manual, many scientists quickly saw its meticulous scanning of both visible and ultraviolet light, and purchased one of their own. (Blauch, 2000)