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Biology AS OCR

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  1. 1-1-practical-skills-written-assessment AS
    7 主题
  2. 1-2-practical-skills-endorsement-assessment AS
    16 主题
  3. 2-1-cell-structure AS
    9 主题
  4. 2-2-biological-molecules AS
    17 主题
  5. 2-3-nucleotides-and-nucleic-acids AS
    8 主题
  6. 2-4-enzymes AS
    9 主题
  7. 2-5-biological-membranes AS
    9 主题
  8. 2-6-cell-division-cell-diversity-and-cellular-organisation AS
    11 主题
  9. 3-1-exchange-surfaces AS
    7 主题
  10. 3-2-transport-in-animals AS
    12 主题
  11. 3-3-transport-in-plants AS
    11 主题
  12. 4-1-communicable-diseases-disease-prevention-and-the-immune-system AS
    16 主题
  13. 4-2-biodiversity AS
    10 主题
  14. 4-3-classification-and-evolution AS
    15 主题
课 6, 主题 9
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2-4-9-practical-measuring-enzyme-activity as

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Exam code:H020

Practical: Measuring Enzyme Activity

  • The progress of enzyme-catalysed reactions can be investigated by:

    • Measuring the rate of formation of a product

    • Measuring the rate of disappearance of a substrate

Investigating catalase activity

  • In this investigation, the rate of product formation is used to measure the rate of an enzyme-controlled reaction:

    • Hydrogen peroxide is a common but toxic by-product of metabolism

    • This means it must be broken down quickly

    • Catalase is an enzyme found in the cells of most organisms that breaks down hydrogen peroxide into water and oxygen

    • Hydrogen peroxide and catalase are combined and the volume of oxygen generated is measured in a set time

    • The rate of reaction can then be calculated

Catalase experiment, downloadable AS & A Level Biology revision notes

Experimental set-up for investigating the rate of formation of a product using catalase

Investigating amylase activity using iodine

  • In this investigation, the rate of substrate disappearance is used to compare rates of reaction under different conditions

  • Amylase is a digestive enzyme that hydrolyses starch into maltose and glucose

  • Amylase functions best at pH 7 and 37oC (all enzymes operate best under specific conditions)

  • Amylase and starch are combined and this reaction mixture is then tested for starch at regular time intervals

  • This can be done by taking samples from the reaction mixture at each time interval and adding each sample to some iodine in potassium iodide solution

    • Starch forms a blue-black colour with this solution

    • If no starch is present, the iodine solution remains yellow-brown

  • In this way, the time taken for starch to be broken down can be measured

  • The investigation can be repeated under a variety of conditions (eg. by altering pH, temperature, enzyme concentration or starch concentration) and the reaction rates can then be compared

Amylase experiment, downloadable AS & A Level Biology revision notes

Experimental set-up for investigating the rate of disappearance of a substrate using amylase

Investigating the effect of starch concentration on amylase activity using colorimetry

  • A colorimeter is able to measure light absorbance (how much light is absorbed) or light transmission (how much light passes through) a substance

  • Colorimetry can be used in any enzyme-catalysed reaction that involves colour change

  • As the colour breaks down the transmission increases or light absorption decreases and this can be used to measure the rate of the reaction

  • For example, a colorimeter can be used to follow the progress of a starch-amylase catalysed reaction as the amylase breaks the starch down into maltose

  • This can be carried out as follows:

    • Colorimeter calibration: this is an important step in a colorimetric investigation and in this case, a weak iodine solution can be used to calibrate the colourimeter as the endpoint (or 100% transmission)

    • Preparation of a starch solution of known concentration (stock solution), from which a range of concentrations are made using serial dilutions (method outlined in diagram below)

    • Following calibration and switching on the red filter (to maximise the percentage transmission or absorbance), the colorimeter is used to measure the percentage absorbance or percentage transmission values

    • Sometimes a reagent or indicator is used to produce the colours detected by the colorimeter and sometimes the solutions themselves absorb light waves

    • A calibration graph is then plotted of starch concentration (X-axis) vs percentage absorbance or percentage transmission (Y-axis)

Serial dilutions, downloadable AS & A Level Biology revision notes

Serial dilution of starch to make a range of concentrations

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