Paul Goddard Core Practical Title: Enzyme concentration and rate of reaction Specification statement:

9 Describe how enzyme concentrations can affect the rates of reactions and how this can be investigated practically by measuring the initial rate of reaction.

Criteria to be assessed: (a)(i) Handles apparatus and materials correctly and safely (b)(i) Measures and observes precisely and records in a structured manner, identifies variables and justifies validity and reliability of results c)(i) Uses appropriate methods to analyse results, present data and identify trends or patterns The sections of the criteria below in bold relate to that section of the report Introduction Scientific Background Amylase is an enzyme that is present in both saliva and pancreatic juice. Its function is to catalyse the hydrolysis of amylose and amylopectin to a mixture of products including maltose and dextrin. Maltose consists of two alpha glucose residues joined by a 1,4 linkage; dextrin is made up of several alphaglucose units joined by both 1,4 and 1,6 linkage. Iodine is a chemical that can react with starch, producing a dark violet colour, which can be used as an indicator. Hypothesis As the concentration of the enzyme increases we would expect to see a decrease in the time taken to reach the achromatic point because as the concentration of the enzyme increases it takes less time to break down the substrate of the starch so it no longer gives a black-blue colour when iodine is added. Method Criteria (b)(i) Measures and observes precisely and records in a structured manner, identifies variables and justifies validity and reliability of results Variables Independent variable- The amount of Amylase added to the starch solution. Dependant variable- The time taken for the achromatic point to be reached Control variables- Volume of enzyme - Volume of iodine added - Time periods between testing - Heat of the water bath Procedure 1. We prepared four different concentrations of the enzyme solution: undiluted diluted to a half, diluted to a quarter and diluted to one tenth of the original concentrations. 2. We then set up a water bath at 35oC. 3. We then used a pipette to put 5cm3 of the undiluted enzyme solution into one test tube and 5cm3 of starch solution into another test tube. 4. We then left both test tubes standing in the water bath and left them for several minutes so they reached the temperature of the water bath 5. The enzyme and starch solution were then mixed together and put them back in the water bath and started a stopwatch. 6. After every minute we removed some of the mixture and mixed it with iodine on a white tile. 7. We then kept taking drops until the mixture failed to give a black-blue colour when, mixed with

iodine, this time was recorded as the achromatic point. 8. We then repeated this method with each concentration, always using 5cm3 of starch and the enzyme. How will reliability and validity of results be ensured? We ensured Reliability as we repeated each concentration a minimum of three times. In addition to this we compared our results with other groups. We ensured that the results were valid by controlling as many variables as we could so that our dependant variable changed as a result of us changing the independent and had minimal influence from other factors. We did this by controlling the volume of enzyme, the volume of iodine added, the time periods between testing and the heat of the water bath.

Results Criteria (b)(i) Measures and observes precisely and records in a structured manner, identifies variables and justifies validity and reliability of results c)(i) Uses appropriate methods to analyse results, present data and identify trends or patterns

Results Table
% Concentration of Enzyme Trial 1
0.02 0.05 0.1 0.2 480 240 120 90

Trial 2
540 420 240 90

Trial 3
600 300 180 90

Trial 4
540 360 240 60

Trial 5

Trial 6
480 240

Trial 7 Average
500 310 150 90 508 290 190 85

210 90

60 30

Graph

Achromatic Point
600 550 500 450 400 350 300 250 200 150 100 50 0 0 0.02 0.05 0.1 0.2

Time (s)

Achromatic Point

Anomalies Anomalies are highlighted in the results table and are most likely caused by human error such as timing issues and incorrect measurements such as; wrong concentrations of enzyme or wrong volumes of starch/enzyme. Random Errors The anomalies highlighted in the table are random errors because they dont reoccur at each concentration and are randomly spread across all of the different trials. Systematic Errors There are no systematic errors because there are not any reoccurring errors that are predictable. Conclusion Criteria c)(i) Uses appropriate methods to analyse results, present data and identify trends or patterns Trends and Patterns of data The average time for each concentration to reach its achromatic point decreases as the concentration, between the first two concentrations, 0 and 0.02 the decrease in time is small however from 0.02 to 0.1 it is a more rapid decrease and 0.1 to 0.2 is also a smaller decrease. At the lowest concentration the time taken to reach the achromatic point is an average of 567 seconds, at the highest concentration it takes 85 seconds to reach the achromatic point. Conclusions with scientific background We concluded that as the concentration increases the time taken to break down the starch substrate decreases because there is more of the enzyme so it can be broken down quicker. There is clear evidence of a decreasing time in the results as at the lowest concentration the time taken to reach the achromatic point is an average of 567 seconds, at the highest concentration it takes 85 seconds to reach the achromatic point.

Teacher Use only Criteria (a)(i) Handles apparatus and materials correctly and safely Achieved / Partially achieved / not achieved

(b)(i) Measures and observes precisely and records in a structured manner, identifies variables and justifies validity and reliability of results c)(i) Uses appropriate methods to analyse results, present data and identify trends or patterns

Other Comments Section Introduction

Comments

Plan / Method

Results

Conclusion and Evaluation