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Yoghurt investigation
The aim of this experiment is to investigate the effect of temperature on yoghurt production by measuring the change in the pH of milk - before and after the starter culture is added.

Yoghurt is made by the fermentation of lactose (milk sugar) by enzymes from bacteria called Lactobacillus bulgaricus. This process is anaerobic, which means that it occurs in the absence of oxygen. During the production of yoghurt, the lactose is broken down by the lactase enzyme (provided by the bacteria) into lactic acid and energy, Lactose lactic acid + energy

The production of lactic acid causes the yoghurt to have a sour taste. Lactic acid also lowers the pH of the milk which affects the proteins in the milk (casein) causing it to coagulate and precipitate, forming the solid curd that makes up yoghurt. The leftover watery liquid is the whey. Before starting making yoghurt the milk is heated to around 70C to kill any other bacteria in the milk, which might also ferment it, so that the only bacteria it contains are the ones we will add. The milk is left to cool, so that the bacteria we are going to add are not also killed, and then the starter culture is added to it.

Dependent variable (what I will measure) Independent variable (what I will change) Controlled variables (what I will keep the same)

range = 2.77 (6.82-4.05=2.77)

temperature: 4 ,20, 30, 40C

volume of milk = 20cm volume yoghurt= 1cm type of milk type of yogurt time = 72h

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Wear a lab coat & goggles to protect you from spills Do not eat the yoghurt

1. With a marker label 4 test tubes 4x, 20x, 30x & 40x and 4 other test tubes 4y, 20y, 30y, 40y. 2. Measure 20cm3 of milk with a measuring cylinder & pour it into each test tube. 3. Measure 1cm3 of yoghurt (starting culture) with a pipette & add it to each test tube. 4. Put a bung in each test tube & shake it gently. 5. Measure the pH of mixture in each test tube with a pH sensor & record it on the results table. Have a beaker with water to rinse the probe each time. 6. Cover each test tube with cling film so no bacteria get in it. 7. Place the 2 test tubes labelled 4x & 4y in the fridge, the 2 labelled 20x & 20y at room temperature, the 2 labelled 30x & 30y in a water bath at 30C and the other 2 labelled 40x & 40y in a water bath at 40C. 8. After 72h observe the mixture in each test tube (if it is liquid or solid) & measure again the pH of mixture in each test tube. Record your observations & measurements on the results table.


Test tube

1cm3 yoghurt (staritng culture) 10cm3 milk

pH sensor

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Results tables
Group1 Group2 Madalena, Francisca, Andr, Toninho, Lus Michael Group3 Joana, Catarina, Rita, Antnio Group4 Benedita, Helena

pH before adding yoghurt

pH after adding yoghurt

Group1 4x 4y 20x 20y 30x 30y 40x 40y 6.43 6.62 6.61 6.68 6.51 6.61 6.53 6.53 6.43 liquid 6.62 liquid 6.61 liquid 6.68 liquid 6.51 liquid/solid 6.61 liquid/solid 6.53 solid 6.53 solid

Table 1 Results from group1

pH before adding yoghurt

Group1 Group2 Group3 6.65 6.75 6.70 6.50 6.51 6.65 6.82 6.69 Group4 6.40 6.20 6.47 6.20 5.97 6.35 6.18 6.42

pH after adding yoghurt

Group1 6.33 6.62 6.54 6.46 4.41 4.42 4.14 4.12 Group2 6.51 6.52 6.26 6.16 4.35 4.41 4.06 4.05 Group3 6.75 6.77 6.55 6.46 4.75 4.73 4.49 4.45 Group4 6.45 6.05 6.22 6.22 6.53 4.22 4.17 4.14

4x 6.43 6.55 4y 6.62 6.55 20x 6.61 6.55 20y 6.68 6.49 30x 6.51 6.47 30y 6.61 6.51 40x 6.53 6.57 40y 6.53 6.78 Table 2 Results from the 4 groups

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pH change = e.g. Group1 6.33-6.43= -0.10 Total = Add all the pH change (x+y) for each temperature e.g. -0.10 + -0.04 + 0.1 + 0.05 +0.00 +-0.03 + 0.02 + -0.15 = -0.15 Mean = e.g. -015/8 = -0.019

pH change
Group1 Group2 Group3 Group4 4x 4y 20x 20y 30x 30y 40x -0.10 0.00 -0.07 -0.22 -2.10 -2.19 -2.39 -0.04 -0.03 -0.29 -0.33 -2.12 -2.10 -2.51 0.10 0.02 -0.15 -0.04 -1.76 -1.92 -2.33 -2.24 0.05 -0.15 -0.25 0.02 0.56 -2.13 -2.01 -2.28 Total for each temperature -0.15 -1.33 -13.76 -18.90 Mean for each temperature -0.019 -0.166 -1.720 -2.363

40y -2.41 -2.73 Table 3 pH change & means

Effect of temperature on yoghurt production

0.500 0.000 0 -0.500 5 10 15 20 25 30 35 40

pH change

-1.000 -1.500 -2.000 -2.500 -3.000

Temperature (C)

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From my results I can conclude that the optimum temperature to produce yoghurt is 40C. At 40C the pH change is the biggest (-2.363), more acidic, the milk coagulates and the mixture gets the texture of yoghurt. For temperatures of 4C and 20C the pH change is very small and the mixture is still liquid, and for 30C although the pH change is -1.720 the milk is not totally coagulated as the mixture is liquid and solid (not the texture of yoghurt). The Lactobacillus bulgaricus present in the starting culture (yoghurt) uses the lactose from the milk as an energy source and the lactase enzyme (provided by the bacteria) produces lactic acid as its fermentation product which lowers the pH of the milk causing it to coagulate. At low temperatures such as 4 and 20C the activity of the lactase enzyme is almost none and as lactic acid is not produced the pH of the milk is not lowered and the milk doesnt coagulate. At 30C the activity of the lactase enzyme is increased, however this activity is not fast enough to ferment the milk and produce enough lactic acid to lower the pH of the milk to the optimum value and coagulate it into the proper texture of yoghurt. A temperature of 40C encourages the growth of bacteria which allows the lactase enzyme to rapidly break down the lactose in the milk into lactic acid. This causes the pH of the milk to decrease to the optimum value which in turn causes the protein from the milk (casein) to coagulate, forming the solid curd that makes up yoghurt.

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