AS Biology Practical - Planning Exercise

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Biology Practical Examination 1 (Part A – Planning Exercise)

Aim: To investigate the effect of the concentration of calcium ions on the coagulation of milk.

Scientific Knowledge and Prediction[1][2][3][4][5]

The major proteins in milk are the caseins; α-caseins, β-caseins and κ-caseins which a group of small phosphoproteins that are able to easily form aggregates (called sub-micelles (see Fig 1.1)). In the presence of calcium ions, these sub-micelles can together form a larger aggregate called the casein micelles (see Figs 1.2 and 1.3). The enzyme rennin binds with κ-casein, forming an enzyme-substrate complex [3], hydrolysing a peptide bond and thus splitting it into two fragments (see Fig 2). The fragment that does not remain part of the sub-micelle (the macropeptide) carries the carbohydrate units[1]; the loss of the carbohydrate ‘coats’ means that strong cross-links between micelles can be formed which is essentially the process of coagulation [1][4] (see Fig 3). Notice how all bonds between sub-micelles in a micelle contain calcium (Fig 1.2). Therefore without calcium ions, no bonds can be made between sub-micelles meaning that micelles can not be formed. Even if rennin did act on the κ-casein, no effect would take place due to the lack of micelles in the first place.

Fig 1:


Fig 2:

Fig 3:

Hypothesis: As long as there is plenty of substrate available, if the calcium chloride solution concentration increases then the rate of coagulation also increases because there will be an increased number of micelles formed. However if we go on increasing the calcium chloride concentration, keeping the volume and concentration of proteins in the milk constant, there comes a point where the calcium ions are in excess and are not contributing to the number of micelles formed. Therefore the rate of coagulation is not increased and will not increase any further.

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Thus the following prediction is made for the graph:

Preliminary Results

Using the method previously described (with equal number of volumes for a single test) the following results were obtained. (Tests were done twice for a given concentration to ensure that they were repeatable.)

The preliminary confirmed the following:

  • The control of 0.0M concentration of the calcium chloride solution confirmed that with the absence of calcium ions, coagulation does not occur on a visible timescale.
  • The points between 0.5M and 1.0M will determine whether the trend is a straight line or a curve, ...

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