lab 6 enzymes introducilon chemica eactions invclve the breaking and formation of bonds enzymes

1. Introduction: Enzymes are organic catalysts that speed up chemical reactions by decreasing th

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LAB 6 ENZYMES INTRODUCTION: Chemical reactions involve the breaking and formation of bonds. Enzymes are organic catalysts; they speed up rates of chemical reactions and are not themselves consumed in the reaction. This is accomplished by decreasing the activation energy (Ea) for chemical reactions. In an enzyme-catalyzed reaction, the two substrates (reactants) are the molecules before the reaction occurs that the enzyme acts upon, and the products are the molecules produced when the reaction is completed. SUBSTRATE + ENZYME -> PRODUCTS Enzymes are involved in both anabolic (building) and catabolic (breaking down) metabolic pathways. The activity of an enzyme can be expressed as the rate at which the enzyme catalyzes the reaction, i.e., how much product is produced per unit of time. In this laboratory, the substrate is sucrose, a disaccharide composed of glucose and fructose. The enzyme sucrase breaks the bond between the glucose and fructose to make the products glucose and fructose by the process of hydrolysis. SUBSTRATE + ENZYME -> PRODUCTS: SUCROSE + SUCRASE -> GLUCOSE + FRUCTOSE (MONOSACCHARIDES) For this lab, sucrase activity will be measured by generating the product glucose, which will be measured with glucose test strips. Sucrase activity can be expressed as how much glucose is produced per unit of time for the reaction, e.g., glucose/5 minutes. The sucrase assay must include the following controls: Negative controls include substrate without enzyme and enzyme without substrate. Neither negative control should result in the production of the product glucose. The positive control is enzyme plus substrate, which should result in the generation of the product. The activity of enzymes may be affected by a number of factors, including the temperature of the reaction, pH of the reaction, substrate concentration, and enzyme concentration. SPECIFIC INSTRUCTIONS: Use the following chart to set up the lab. The chart indicates how many drops of each substance to add to each tube. For tubes 1-15, make additions to the tubes except for the sucrase. After all additions have been made for tubes 1-15, then add the sucrase to each tube. This should be done as quickly as possible. Begin timing the reaction after the addition of the sucrase. After 5 minutes of reaction, immerse the colored end of one glucose test strip into each tube, wetting the strip and then removing it from the tube. Approximately 1 minute after the strip was wet, read the results by comparing the color of the pad of the strip with the chart on the strip container. Be sure to read all strips being compared in the sequence they were set up and at the same time. Record the results immediately since the color will continue to change as the strip sits in the air. For tubes 15 and 18, a different sequence is employed. First, add the enzyme alone to the tube and then subject the enzyme to the specified conditions (pH solution, temperature, or salt solution). For tubes 12-15, heat the tubes for two minutes at each temperature. Remove the tubes from the refrigerator and water baths before heating.

LAB 6 ENZYMES INTRODUCTION: Chemical reactions involve the breaking and formation of bonds. Enzymes are organic catalysts; they speed up rates of chemical reactions and are not themselves consumed in the reaction. This is accomplished by decreasing the activation energy (Ea) for chemical reactions. In an enzyme-catalyzed reaction, the two substrates (reactants) are the molecules before the reaction occurs that the enzyme acts upon, and the products are the molecules produced when the reaction is completed. SUBSTRATE + ENZYME -> PRODUCTS

Enzymes are involved in both anabolic (building) and catabolic (breaking down) metabolic pathways. The activity of an enzyme can be expressed as the rate at which the enzyme catalyzes the reaction, i.e., how much product is produced per unit of time. In this laboratory, the substrate is sucrose, a disaccharide composed of glucose and fructose. The enzyme sucrase breaks the bond between the glucose and fructose to make the products glucose and fructose by the process of hydrolysis. SUBSTRATE + ENZYME -> PRODUCTS: SUCROSE + SUCRASE -> GLUCOSE + FRUCTOSE (MONOSACCHARIDES)

For this lab, sucrase activity will be measured by generating the product glucose, which will be measured with glucose test strips. Sucrase activity can be expressed as how much glucose is produced per unit of time for the reaction, e.g., glucose/5 minutes. The sucrase assay must include the following controls: Negative controls include substrate without enzyme and enzyme without substrate. Neither negative control should result in the production of the product glucose. The positive control is enzyme plus substrate, which should result in the generation of the product.

The activity of enzymes may be affected by a number of factors, including the temperature of the reaction, pH of the reaction, substrate concentration, and enzyme concentration.

SPECIFIC INSTRUCTIONS: Use the following chart to set up the lab. The chart indicates how many drops of each substance to add to each tube. For tubes 1-15, make additions to the tubes except for the sucrase. After all additions have been made for tubes 1-15, then add the sucrase to each tube. This should be done as quickly as possible. Begin timing the reaction after the addition of the sucrase. After 5 minutes of reaction, immerse the colored end of one glucose test strip into each tube, wetting the strip and then removing it from the tube. Approximately 1 minute after the strip was wet, read the results by comparing the color of the pad of the strip with the chart on the strip container. Be sure to read all strips being compared in the sequence they were set up and at the same time. Record the results immediately since the color will continue to change as the strip sits in the air.

For tubes 15 and 18, a different sequence is employed. First, add the enzyme alone to the tube and then subject the enzyme to the specified conditions (pH solution, temperature, or salt solution). For tubes 12-15, heat the tubes for two minutes at each temperature. Remove the tubes from the refrigerator and water baths before heating.

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