Alkaline Phosphatase Enzyme Lab Report

The substrate for beta-galalactoside is ortho-nitrophenyl-B-galactoside. ONPG is structured similarly to lactose. The purpose of the experiment was to add a competitive inhibitor to observe if the reaction rate would slow down. A competitive inhibitor is when the inhibitor binds to the active site on the enzyme and prevents the binds of the substrate

Dependent Variable amount of product (glucose and fructose) produced 2. Independent Variable temperature 3. Controlled Variables pH, amount of substrate (sucrose) present, sucrase + sucrose incubation time Effect of Substrate Concentration on Enzyme Activity 1. Dependent Variable amount of product (glucose and fructose) produced 2.

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.

This reaction involves oxidation which is the loss of electrons and reduction which is the gain of electrons, both of which occur simultaneously and depend upon each other. An example of an Oxidoreductase enzyme is Lactate Dehydrogenase. The class Transferase catalyses reactions which transfer functional groups such as amino groups, phosphate groups or others. An example of the Transferase class of enzyme is Alanine Deaminase. Hydrolase catalyses any hydrolysis reaction and examples are Lipase and Sucrase.

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 3.5.3.1 (Schomburg 2015). The enzyme ‘commission number’ is the arithmetical classification that is used for enzymes which indicates the chemical reaction they catalyze.

An enzyme biologically defined is a catalyst produced by cells to speed up specific chemical reactions without changing the chemical reaction at the end of the reaction.1 There are several factors that affect the rate an enzyme speeds up reactions; temperature, pH, substrate concentration and enzyme concentration.2 However, when there is too much or not enough of these factors (depending on the enzyme) it can destroy the enzyme entirely. In this experiment we tested how temperature affects enzymes. We observed the enzyme activity for the enzyme Alkaline phosphatase when it was put in an environment of 33°C and 86°C. Because Alkaline phosphatase has the ability to extract phosphate groups from substrates, once the Alkaline phosphatase was in the specifically heated environment for five minutes, we measured it’s activity by inserting para-nitrophenyphosphate.

4.3) Briefly explain what you understand by specific activity of an enzyme and how you could measure it. (P 4.3) Specific activity is a term utilize to measure the rate of reaction of an enzyme with a substrate. Specific enzyme activity is a measure of enzymes purity and quoted as units/ mg. the value becomes huge as an enzyme preparation becomes purer since the amount of protein (mg) is typically less, but the rate of reaction stays the same/ may increase due to reduced interference/ removal of inhibitors.

In addition, phenolphthalein was added as an indicator. The aliquots were titrated against sodium hydroxide (NaOH) solution until end point was reached, after which volume of NaOH consumed was recorded. The value of the rate constant, k, obtained was 0.0002 s-1. The experiment was then repeated with 40/60 V/V isopropanol/water mixture and a larger value of k = 0.0007 s-1 was obtained. We concluded that the rate of hydrolysis of (CH3)3CCl is directly proportional to water content in the solvent mixture.

Globin- It is a protein surrounding & protecting the heme molecule. Heme synthesis: Heme synthesis is carried out in mitochondria & cytosol of the cell involving cascade of steps :- 1) The first step occurs in mitochondria, where condensation of succinyl-CoA & glycine is carried out by enzyme ALA-synthase resulting in product formation i.e. 5-aminolevulinic acid.

Therefore pyruvate must be oxidised to yield Acetyl-CoA and CO2 which is carried out by pyruvate dehydrogenase (PHD). This is a complex structure that consist of a cluster of enzymes found in the mitochondria of eukaryotic cells. This reaction is called the oxidative decarboxylation. It is an irreversible oxidative process. Here the carboxyl group is removed from the pyruvate as a molecule of C02 and the remaining two carbons are used to become the acetyl group in the Acetyl-CoA. Therefore pyruvate C3 is converted to acetate c2.

1. The first step of my calculations was finding the number of moles of CaCl2 and NaOH added in each test. The volume of CaCl2 is an increasing number with a concentration of 1.0M. The volume of NaOH is constant for all four tests, but the concentration is 2.4588M. To find the number of moles of each reactant added, volume in liters was multiplied by the molarity (concentration). 2.

The motivation of this investigation was to achieve 85% of methanol recovery from the distillate. II. Methodology: The distillation column was analyzed theoretically using McCabe Thiele to establish the number of stages required for separation. The vapor-liquid equilibrium (VLE) data for methanol and 2-propanol was used to plot curves of methanol-vapor fraction versus methanol-liquid fraction, and methanol liquid-vapor fraction versus temperature.