INTRODUCTION: Arginase is an enzyme- enzymes are biological catalyst which drives a reaction at the speed of life. Arginase is a hydrolase, hydrolases catalyze hydrolysis reactions, this is determined via the E.C number (Nelson and Cox 2008). Arginase has the EC number is 188.8.131.52 (Schomburg 2015). The enzyme ‘commission number’ is the arithmetical classification that is used for enzymes which indicates the chemical reaction they catalyze. EC 3 are hydrolases, which forms two products from the substrate via hydrolysis.
Analysis is performed when equilibrium is obtained. In this circumstance, it refers to the association rate and dissociation rate of complex formation being equal. The fractional occupancy at equilibrium is predicted by the law of mass action Firstly, a protein standard curve will be created to determine the concentration of the unknown sample. When this is completed, the saturation binding graph will be created using graph pad prism and determine the Bmax and Kd values. A Scathard plot will then be created and the Bmax obtained from the x-axis intercept and the Kd value obtained from the slope of the line the.
After filtration, it was centrifuged. The supernatant was then isolated and used for fractionation. Fractionation: Precipitation of proteins from a solution may be accomplished by fractional precipitation techniques using salts, organic solvents or pH. The first method of protein precipitation was by using salts. Sodium phosphate with pH 7 was used.
The inverted solution C is used to carry out another titration with Fehling’s solution to determine the concentration of reducing sugars as invert after hydrolysis, where all sucrose are hydrolyzed into glucose and fructose. By carrying out two titrations, we can therefore determine the concentration of reducing sugars obtained from the hydrolysis of sucrose by finding the difference between the concentration of reducing sugars of both titrations. One molecule of sucrose is made up of one molecule of glucose and one molecule fructose joined by 1,2-glycosidic linkage. Therefore, the concentration of reducing sugars obtained from the hydrolysis of sucrose is multiplied with a factor of 0.95 (the molecular weight of sucrose which is 342 g/mol to the total molecular weight of glucose and fructose which is 360 g/mol). The value obtained from this step is the concentration of sucrose present in the strawberry jam sample, which is
ABSTRACT To catalyze a reaction, an enzyme will grab on (bind) to one or more reactant molecules. In this experiment we examined how increasing the volume of the extract added to the reaction would affect the rate of the reaction. The enzyme used was horseradish peroxidase which helps catalyze hydrogen peroxide. Using different pH levels, the absorbance rate of the reaction was measured to see at which condition the enzyme worked best. The rates of absorption were calculated using a spectrophotometer in 20 second intervals up to 120 seconds.
Sodium dodecyl sulfate polyacrylamide gel electrophoresis also known as SDS-PAGE is one of the methods for determining the molecular weight of unknown proteins. SDS is an anionic molecule which denaturizes proteins and brings it back to its’ primary structure and it also provides a negative charge to the uncharged molecule. The SDS-PAGE enables the separation of proteins based on their sizes. The larger the size of the protein, the harder it is to travel through the gel thus heavier proteins stay near the cathode side of the gel. For this experiment, a software named Gel Analyzer was used in order to obtain the molecular weight of the unknown proteins with the help of a protein ladder with known molecular weight and protein concentration.
Introduction 1.1 Aim: To determine the kinetic parameters, Vmax and Km, of the alkaline phosphatase enzyme through the determination of the optimum pH and temperature. 1.2 Theory and Principles (General Background): Enzymes are highly specific protein catalysts that are utilised in chemical reactions in biological systems.1 Enzymes, being catalysts, decrease the activation energy required to convert substrates to products. They do this by attaching to the substrate to form an intermediate; the substrate binds to the active site of the enzyme. Then, another or the same enzyme reacts with the intermediate to form the final product.2 The rate of enzyme-catalysed reactions is influenced by different environmental conditions, such as: concentration
Lab Report -- Relationship on Enzyme activity and substrate concentration Research Question: Is the more concentrated the substrate of hydrogen peroxide is, the shorter the time taken for the paper disc to rise from the bottom of the beaker? Aim: The opposite of hull hypothesis Background Information: This experiment aimed to investigate on the relationship of the substrate concentration and enzyme activity. Enzymes are proteins produced by a cell that acts as catalysts to increase the rate of a specific chemical reaction without changing the reaction itself. Under some conditions, substrate will bind to the active site of an enzyme and form an enzyme-substrate complex. The enzyme would fasten the chemical reaction and the substrate will
Literature review Research question is how different temperatures affect the catalase enzyme. What is an enzyme? Enzymes are macromolecular biological catalysts. Enzymes speed up chemical reactions. Substrates are molecules that enzymes could act upon and the enzyme converts the substrates into different molecules known as products.