The Effect Of Alkaline Phosphatase On Para Nitrophenylphosphate

In this lab or experiment, the aim was to determine the following factors of enzymes: (1) the effects of enzymes concentration the catalytic rate or the rate of the reaction, (2) the effects of pH on a particular enzyme, an enzyme known and referred throughout this experiment as ALP (alkaline phosphate enzyme) and lastly (3) the effects of various temperatures on the reaction or catalytic rate. Throughout the experiment 8 separate cuvettes and tubes are mixed with various solutions (labeled as tables 1,3 & 4 in the apparatus/materials sections of the lab) and tested for the effects of the factors mentioned above (concentration, pH and temperature). The tubes labeled 1-4 are tested for pH with pH paper and by spectrophotometer, cuvettes 1a-4a was tested for concentration and cuvettes labeled 1b-4b was tested for temperature in four different atmospheric conditions (4ºC, 23ºC, 32ºC and 60ºC) to see how the enzyme solution was affected by the various conditions. After carrying out the procedures the results showed that the experiment followed the theory for the most part, which is that all the factors work best at its optimum level. So, the optimum pH that the enzymes reacted at was a pH of 7 (neutral), the optimum temperature that the reactions occurs with the enzymes is a temperature of 4ºC or

Five cuvettes provided by the instructor were used during the experiment. The transmittance on the spectrophotometer was zeroed on an empty chamber and it was set at a wavelength of 486 nm. There was only one blank containing 0.5 mL of enzyme solution (catechol oxidase) and 4.5 mL of pH 6 buffer. The blank was prepared at the beginning of the experiment and used throughout the experiment. The blank was used to zero the absorbance on the spectrophotometer before each experimental trial. The experimental cuvettes contained 0.5 mL of catechol oxidase (enzyme), 0.5 mL of 5mM catechol (substrate), and 4.0 mL of pH 6 buffer for a total of 5.0 mL of solution. The results for the experimental trials were obtained after 0 seconds, 30 seconds, 1minute and 30 seconds, 3 minutes, and 5 minutes in five different reactions. The enzyme and substrate were obtained using a micropipette, and the pH was measured using a 10mL pipet. Each experimental trial was carried out twice for a total of ten experimental

The practical was carried out to investigate the effect of pH on the reaction of the enzyme acid phosphatase.

These results shown from this experiment led us to conclude that enzymes work best at certain pH rates. For this particular enzyme, pH 7 worked best. When compared to high levels of pH, the lower levels worked better. The wrong level of pH can denature enzymes; therefore finding the right level is essential. The independent variable was the amount of pH, and the dependent being the rate of oxygen. The results are reliable as they are reinforced by the fact that enzymes typically work best at neutral pH

The purpose of this lab is to understand how non-living systems can make it easier for chemical reactions to occur. In this lab, raw wheat germ acid phosphate was used to detach it from other chemicals. Raw wheat germ is an example of an enzyme which is defined as, “a biological catalyst, all catalysts work by speeding up the rate at which a chemical reaction occurs.” (Krane, 2018). Enzymes are made up of proteins that can be isolated in two different ways, assay and precipitation. Assay is defined as, “generally readily available for most enzymes and depend on the ability to detect a change (often in color) as substrates are converted to products as the reaction occurs.” (Krane, 2018). Precipitation can be described as, “procedures often begin with a series of trial and error experiments. There is no standard protein isolation technique that can be applied to all proteins and some large classes of proteins, like membrane-bound proteins.” (Krane, 2018). This lab was performed in three different sections, the first one being wheat germ suspension, where heat was used to purify and isolate the wheat germ. In the second portion of the lab, examiners found the concentration of the wheat germ protein. In the final step of the experiment examiners found the amount of acid phosphatase in each of the tested microtubes.

The objective of this lab was to develop a protocol to investigate the effect of an environmental variable on the catalytic function of an enzyme. More specifically, the objective was to perform an experiment in order to test the effect of pH on the function of the enzyme catalase.

J. Moldovan & B. Nilson, (2010), Lab 4 – Enzyme Kinetics, UBCO BIOL/BIOC 393, UBC Vista accessed Monday, November 8th, 2010.

Alkaline phosphatase will behave like a protein, due to the fact that it is a glycoprotein. Utilizing the above mentioned information it is possible to test for the optimum pH. Alkaline phosphatase will function optimally at a pH of around 10 (an alkaline pH). If one was to assess the range of the pH pertaining to this enzyme, it can be seen that there will be a point where the enzyme has the greatest activity, whereby it produces the highest concentration of the so-called “product”. Due to the previous knowledge of proteins, it can be deduced that at acidic pH the protein will have very low activity or will denature. It should also be noted that once the optimal pH has been reached and surpassed, the enzymes rate of activity will begin to reduce.

BI532: Skills for Bioscientists 2 Mini Project 2 report Effect on the velocity Alcohol Dehydrogenase on different alcohols under different conditions Abstract The effect on the velocity of Alcohol dehydrogenase producing an acetaldehyde was measured under different conditions to analyse the properties of the enzyme as well as the concentration the enzymes have the best rates. The velocity of alcohol dehydrogenase was measured against different substrates such as ethanol, propanol and propan-2-ol and using spectrophotometry at a wavelength of 340nm. While the enzyme velocity of ethanol production was high, propan-2-ol did not cause any changes the rate of acetaldehyde production. The amount of ethanol, which was the most reactive substrate, that reacted with the fastest rate was then calculated

An enzyme is a protein that acts as a catalyst which reduces the activation energy needed for a chemical reaction. Without the presence of enzyme, cell reactions would take so long that they would detectable. During a reaction, in the presence of an enzyme, the substrate first creates a complex with the enzyme. While the substrate is a part of the complex, it’s converted into the product. Then, finally, the complex dissociates from the molecule which allows the release of the enzyme and formed product. An enzyme’s activity depends on a variety of conditions which includes the pH level and temperatures. Phosphorylase is an enzyme that catalyze the addition of a

Enzymes are an important part of all metabolic reactions in the body. They are catalytic proteins, able to increase the rate of a reaction, without being consumed in the process of doing so (Campbell 96). This allows the enzyme to be used again in another reaction. Enzymes speed up reactions by lowering the activation energy, the energy needed to break the chemical bonds between reactants allowing them to combine with other substances and form products (Campbell 100). In this experiment the enzyme used was acid phosphates (ACP), and the substrate was p-nitrophenyl phosphate.

There is a two-step reaction for hydrolyzing phosphate monoester by PTPs. First, there is a nucleophilic attack of the cysteine (Cys) in the active site on the phosphorus atom of the target substrate. At the same time as the ester bond is cleaved, the aspartic acid (Asp) residue donates its proton to the leaving group oxygen. This reaction leaves the phosphate group covalently attached to the nucleophile via a thioester linkage. In the second step, the phospho-enzyme intermediately is hydrolyzed by a water molecule which gives the enzyme and inorganic phosphate. The conserved Asp in the first step acts as a general base to activate the water molecule and helps to hydrolyze the intermediate complex

Acid phosphatase: Variation of enzyme-catalyzed reaction rate with time and effect of enzyme concentration on p-nitrophenol Aim To investigate the change in reaction rate of p-NPP throughout an extended period of time and the effect of acid phosphatase concentration on the initial rate of the enzyme-catalyzed reaction using stopped assays. Method(Coordinators, 2016) Three experiments were carried out in the entire investigation in order to obtain a standard curve of p-nitrophenol construction, the time course of the formation of p-nitrophenol, p-NP, from p-nitrophenylphosphate, p-NPP, and another experiment with various concentrations of acid phosphatase reacting with p-NPP. All experiments were performed in a microtitre plate at room temperature, around 25 ℃. The absorbance of the wells for each experiment were simultaneously read at 405nm with the spectrophotometer.

pH levels effect on Enzyme Activity and Concentration Abstract: pH level may affect enzyme activity and the performance of the enzyme which is crucial in order for the enzyme to properly function in our biological systems. We investigated the relationship between pH level and rate of enzyme function at that certain pH level. The enzyme concentration was recorded at varying levels of pH using the enzyme Lactase, substrate ONPG, and phosphate buffer in a spectrophotometer set at wavelength 405nm. As pH levels became extremely high or low, enzyme concentration decreased.

The purpose of this lab report is to investigate the effect of substrate concentration on enzyme activity as tested with the enzyme catalase and the substrate hydrogen peroxide at several concentrations to produce oxygen. It was assumed that an increase in hydrogen peroxide concentration would decrease the amount of time the paper circle with the enzyme catalase present on it, sowing an increase in enzyme activity. Therefore it can be hypothesised that there would be an effect on catalase activity from the increase in hydrogen peroxide concentration measured in time for the paper circle to ride to the top of the solution.