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EXPERIMENT 5

EXPERIMENT 5. DESIGN OF EXPERIMENT ABOUT THE BEST CONDITIONS FOR AMYLASE ACTIVITY. 2012-09. Aims. You should get the thread how to design a project about experiment research by the design of experiment and should be able to put it in practice.

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EXPERIMENT 5

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  1. EXPERIMENT 5 DESIGN OF EXPERIMENT ABOUT THE BEST CONDITIONS FOR AMYLASE ACTIVITY 2012-09

  2. Aims • You should get the thread how to design a project about experiment research by the design of experiment and should be able to put it in practice. • You would get deeper insight into some methods of enzyme kinetic research, for example, how does the change of pH affect the reactions catalyzed by enzyme

  3. Project ( in your free time before the experiment ) • Review the contents about the main factors influencing the reactions catalyzed by enzyme. • Consult the research state related to amylase at present, including the functions, secretion, activity determination of amylase and its relationship with some clinic illnesses. You can use the library or/and internet.

  4. Preparation • Two students as a group, to discuss the project of experiment design about amylase. • You have to decide the concentration of the enzyme or substrate as well as reaction time about the experiment. • You should find the best pH buffer for the activity of saliva amylase by putting your experiment project in practice.

  5. Human salivary amylase: calciumion visible in pale khaki, chloride ion in green.

  6. How to do • Prepare for some experiment conditions, such as reagents, instruments, spectrophotometer 721. • Write down the steps how to get the saliva amylase, which diluted multiple of saliva amylase should be used. • Hand in your project to your demonstrator before the experiment. Get some advise about the procedures from your demonstrator.

  7. Protocol ( two students as a group ) • Read your project carefully again in order to understand clearly the significance and the aims of the experiment design. • Get saliva amylase by yourself and carefully considered dilute it. • Make up the diverse pH for the experiment.

  8. Protocol • Measure the activity of saliva amylase in different pH solution. Draw the graph according to the experiment data in order to find the best pH for the saliva amylase. • Amend the experiment conditions such as amylase concentration, substrate concentration and their amount in the reaction until getting appropriate data to determine the best pH of amylase.

  9. Activity of enzyme pH

  10. A660 pH

  11. Discussion • Which pH buffer is the best for the activity of saliva amylase? Why? • Discuss the outcome of the experiment each other, write down a short summary about the experiment design.

  12. Next Week: Review The application of enzyme knowledge in clinic, such as diagnosis, treatment, and prevention Title • Author, address • Abstract • Key word • Introduction • Formal contents • Reference Format

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