Horseradish Peroxidase Lab Report

Enzymes speed up chemical reactions enabling more products to be formed within a shorter span of time. Enzymes are fragile and easily disrupted by heat or other mild treatment. Studying the effect of temperature and substrate concentration on enzyme concentration allows better understanding of optimum conditions which enzymes can function. An example of an enzyme catalyzed reaction is enzymatic hydrolysis of an artificial substrate, o-Nitrophenylgalactoside (ONPG) used in place of lactose. Upon hydrolysis by B-galactosidase, a yellow colored compound o-Nitrophenol (ONP) is formed.

The effect of pH on the speed of enzyme interaction with substrate chemicals Hypothesis: About pH: If the pH level is less than 5, then the speed of the enzyme reaction will be slower. About temperature: If the temperature stays the same, then the speed of the enzyme reaction will not be completely affected. Background information: The function of enzymes is to speed up the biochemical reaction by lowering the activation energy, they do this by colliding with the substrate. All enzymes are under the class of protein biomolecule. Amino acids are the basic units that are combined to make up an enzyme.

The Aim of Enzyme Catalase Experiment is making a series of experiments involving the enzyme Catalase which has been performed in order to determine some of the enzyme 's properties. The enzyme found in different conditions which its specific reaction rate. Variation in enzyme concentration, variation in pH, variation in temperature, and the effect of different concentrations of inhibitors were all tested. The enzyme concentration increased the reaction rate. An optimum pH and temperature were found for the enzyme, outside of this optimum the reaction rate would be lower.

Catalyse Enzyme Experiment. Enzymes are biological catalysts. They speed up chemical reactions which go on inside living things. Without them reactions would be so slow that life would grind to halt. These are examples that can decomposition of the hydrogen peroxide.

Catalase reacts with hydrogen peroxide, binding onto it and breaking it down into the less toxic water and oxygen. The equation for this reaction is the following: 2 H2O2 = 2 H20 + 2 O2 This experiment will use 1% catalase solution and 3% hydrogen peroxide solution, both diluted into water so the reaction slows down. Temperature will be controlled in this experiment to change the reaction speed of the enzyme and the substrate, this is what the experiment is looking at. The effect of the temperature will be determined by how much gas is released in two minutes, which will change the pressure inside the test tube and will be measured by a gas

Enzyme assays are performed to serve two different purposes: (i) To identify a special enzyme by proving its presence or absence in a distinct specimen. (ii) To determine the amount of the enzyme in the sample by monitoring the disappearance of substrate or appearance of product. Enzymes speed up reaction rate by decreasing the activation energy required to start the reaction. Activation energy is the energy required to break certain bonds in the substrate so that other bonds can form. The formation of these new bonds results in the formation of the product by measuring the changes in absorbance due to the substrate (starch) being changed into product by the amylase enzyme.

After the binding of a substrate to an active site, the active site is occupied and unavailable to other substrate molecules until products have been composed and released from the active site (Allott and Mindorff). As the concentration of the substrate rises, more and more of the active sites consequently get occupied. As a result of all of this, a greater proportion of substrate-active site collisions are blocked. For this reason, the rate at which enzymes catalyze reactions gets smaller as substrate concentration increases. Aim of this investigation is to find out how much of an effect there is on enzyme activity and reaction time as pH values change.

Objective: The purpose of this experiment is to determine the changes in activity level (if any) when enzymes are exposed to a variety of environments (in this case, temperature). Introduction: Enzymes are made up of a series of proteins known as amino acids. Enzymes are essential in almost all aspects of life. There are certain ways to identify an enzyme by the name for each, for example they usually end with the suffix -ase. For instance, amylase (present in saliva), lipase, or protease.

Amino acids are the building blocks of proteins. All amino acids have the same basic structure but differ in their R-side chains. Each amino acid consists of an amino group (-NH3), a carboxyl group (-COOH) and a hydrogen atom (H). The amino and carboxyl groups are attached to a central alpha carbon together with a hydrogen atom and an R-side chain. There are currently known that over 170 amino acids occur in organisms but only 20 are commonly found in proteins.

To 0.05 ml of carbonate buffer (0.05 M, pH 10.2) and 0.5 ml of EDTA (0.49 M) was added. The reaction was initiated by the addition of 0.4ml of epinephrine and the change in optical density/min was measured at 480nm. SOD activity was expressed as units/mg protein change in optical density/min. 50% inhibition of epinephrine to adrenochrome transition by enzyme is taken the enzyme unit. Calibration curve was prepared by using 10 -125 units of

purpose the propose of this experiment was too see if the chemical reaction of a enzyme can be made faster. Hypothesis I think that a warm environment would be best to make an enzyme’s reaction faster. because a protein can move faster in heat. Materials -pan -50ml graduated cylinder -hydrogen peroxide -air stopper -water Graphs data A time 12 drops 8 drops 0 0 0 30 0 0.5 60 0 1 90 0 1 120 0 1 150 0 1 180 0 1.5 210 0 1.5 240 0 1.5 270 0 1.5 300 1 1.5 330 2 1.5 360 2 1.5 390 3 1.5 420 3 1.5 450 3 2 480 3 2 510 3 2 540 4 2.5 570 4 2.5 600 5 2.5 Data B time cold warm 0 0 0 30 1 1 60 2 1 90 2 2 120 2 2 150 2 2 180 2 2 210 2.5 2 240 3 2 270 3 2 300 3 2 330 3 3 360 3.5 3 390 3.5 3 420 3.5 3 450 3

1. How does DNA encode information? DNA is a double-stranded helix composed of a phosphate backbone and deoxyribose, and encodes information by the sequence of its nucleotide bases, which are composed of adenine, thiamine, guanine and cytosine. DNA undergoes transcription, which produces single-stranded mRNA, which uses uracil in place of thiamine. Next step is translation, in which the RNA becomes a protein, which then can act as structural units or enzymes.

The constants of the experiment, will be the amount of water used and the Alka Selter compound. The control in the experiment is water. Units used while timing the productivity of gas from an Alka-Seltzer tablet in different temperatures is, seconds. In order to find out if temperature controls the rate of chemical reaction, whether hot water is a more effective way to make the gas produce at a faster speed, it would be necessary to compare the results of different temperatures at the end of each trial. In order to do this the scientists will measure the volume of gas that is produced within a 10 second interval time after the tablet begins to react.

Cellular respiration can be measured by the consumption of oxygen, the consumption of carbon dioxide, and the release of energy during cellular respiration. Within the experiment conducted, the relative volume of O2 consumed was measured into different temperatures within germinating and nongerminating peas, (DeStefano). Fluids and gas flow from regions of high-pressure to regions of low-pressure this carbon dioxide produced during cellular respiration will be removed by potassium hydroxide and will form a solid potassium carbonate. Due to the removal of carbon dioxide, the change in the volume of gas in the respirometer will be directly related to the amount of oxygen consumed. In this experiment using a respirometer, the scientists were able to measure the amount of oxygen being consumed in relation to how quickly the peas were respiring.

The purpose of stacking gel is to make sure all the proteins start separating at approximately the same time. The pore size is larger so that it is easier for large protein to move in order to catch up with the smaller protein. As heating, SDS denature the proteins, the proteins are loaded onto the wells on the stacking gel. The denatured proteins have a uniform mass to negative charge ratio. Since the current run from negative (top) to positive (bottom), the proteins move toward the bottom.