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AS2 Investigation Winter 2002

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Introduction

AS2 Investigation Winter 2002 The Problem In industry, enzymes are often immobilised. There are a number of advantages of this method, it allows the enzyme preparation to be reused, the product obtained is enzyme free and immobilisation offers a degree of thermostability. Task Design an experiment to investigate the statement "immobilisation offers a degree of thermostability when compared to free enzymes". Equipment List Amylase solution Sodium Alginate solution Calcium Chloride solution Starch solution Iodine solution Distilled water 3 Beakers 250cm ) 2 Syringes (10cm ) Measuring cylinder Droppers Spotting tiles Filter paper and funnel Stopclock Water baths at 30?C, 40?C, 50?C, 60?C, 70?C 2 boiling tubes Variables In this experiment there are several possible independent variables that could be changed, these consist of: * The temperature at which the enzyme-substrate reaction takes place * The amount of enzyme (amylase) used * The amount of sodium alginate used * The amount of substrate (starch solution) used * The concentration of the starch solution * The concentration of amylase solution used * The concentration of sodium alginate ...read more.

Middle

The concentration of amylase solution used: To keep this constant, I shall use the same amylase solution in each 4cm syringe uptake that I do. The concentration of sodium alginate solution used: To keep this constant, I shall use the same sodium alginate solution in each 6cm syringe uptake that I do. The dependant variable that I will measure will be the time it takes for all the starch to be broken down. To get a set of quantative results, the time at which the iodine goes deep brown will be measured, in both free and immobilised enzymes, against the temperature of the water bath. Method The following method will be used to perform the experiment: 1. Collect all equipment. 2. Measure up 50cm of starch solution into each of two clean beakers clearly labelled. 3. Place the two beakers containing the starch solution into a water bath at 30?C. 4. Measure up 50cm of calcium chloride solution into each of two clean beakers, clearly labelled. ...read more.

Conclusion

The droppers must be rinsed with distilled water after each test. Repeat the above process using water baths at different temperatures: 40?C, 50?C, 60?C and 70?C. If starch is still present the iodine will go black/blue. If starch is not present the iodine will go deep brown. To ensure my results are reliable I will do the experiment 3 times. To ensure I work safely I shall wear protective goggles and I shall behave according to GLP (General Laboratory Practice. To make the experiment a fair test, I shall: * Replace the sodium alginate with distilled water in the free enzyme solution. * Use equal amounts of starch in each beaker. * Make sure to the best of my ability that all beads have the same surface area. * Make sure all the 30second timed intervals are exact. * Make sure the same amount of iodine is used in each trough on the spotting tile. * Make sure the same amount of enzyme is used in both free and immobilised mixes. * Make sure the end point reached in the iodine is the same each time. Michael Edwards Biology Miss Dunn ...read more.

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