KYAMBOGO UNIVERSITY

BACHELAR OF SICENCE IN CHEMICAL AND

PROCESS ENGINEERING
YEAR 3
NKUGWA MARK WILLIAM
17/U/7029/CHE/PE
PRACTICAL: MICROBIOLOGY
SIGN:
 MAKING YOGURT

AIM
 The purpose of this experiment is to produce yogurt by adding specific bacteria to
prepared milk.
 To observed and compare between an experimental and commercial yogurt.

INTRODUCTION
In this experiment we were required to produce yogurt by adding specific bacteria to prepared
milk. The product will be observed and comparisons will be made between an experimental and
commercial yogurt.

ABSTRACT
In this experiment using beakers and a hot plate, milk was placed heated indirectly to 90 oC and
not allowed to boil. This was done to remove most of the micro-organizes which are not wanted
in the milk. Sugar was then added to the milk and it was allowed to cool to 32oC so as to allow
the optimal growth of the bacteria to be added to the solution. Bacteria where then introduced
into the solution and oxygen was then cut off from the solution to create anaerobic conditions so
as the bacteria can grow well and enable the fermentation process to take place well. PH was
recorded after an hour 6 times as it was seen that the best pH was 4 to 4.6 for forming good
yogurt with proper tart and thickness. Flavor was then added and the product formed was packed
and compared with commercial yogurt.

THEORY
Yogurt is made by the fermentation of lactose (milk sugar) by bacterial enzymes. Yoghurt is a
dairy product produced by bacteria fermentation of milk sugar (lactose) into lactic acid. This
gives yoghurt its gel-like texture and characteristics taste. It is often sold with a fruit vanilla or
chocolate flavor but can be unflavored. Its nutritional and therapeutic functions have been this
process is anaerobic, meaning that it occurs in the absence of oxygen. Lactose is a compound
sugar, made up of the two simple sugars glucose and galactose. During the making of yogurt, the
lactose is broken down by the lactase enzyme (provided by bacteria) into these two components,
as shown in Figure 1. Further processing of glucose and galactose results in the end products of
lactic acid and acetaldehyde. The production of lactic acid and acetaldehyde lowers the pH of the
milk, causing it to have a sour, tart taste. The lower pH also affects the casein (milk protein),
causing it to coagulate and precipitate, forming the solid curd that makes up yogurt. The leftover
watery liquid is the whey. The two bacteria most commonly used to make yogurt are
Lactobacillus bulgaricus and Streptococcus thermophilus.

THE BIOCHEMISTRY BEHIND YOGURT

Yogurt is a product of the acidic fermentation of milk. The lactose in the milk is converted to
lactic acid, which lowers the pH. When pH drops below pH 5, micelles of caseins, a hydrophobic
protein, loses its tertiary structure due to the protonation of its amino acid residues. The
denatured protein reassembles by interacting with other hydrophobic molecules, and this
intermolecular interaction of caseins creates a structure that allows for the semisolid texture of
yogurt

Fig.3
Yogurt production begins with the breakdown of lactose into glucose and galactose (Fig. 2), a
process catalyzed by β-galactosidase. The glucose produced from this catabolic step then enters
glycolysis, producing pyruvate. It has been proposed that yogurt bacteria utilize the Embden-
Meyerhof-Parnas pathway of glycolysis (Fig. 3). Pyruvate then enters lactate fermentation, also
known as homolactic fermentation, as it produces only lactic acid molecules. In other types of
fermentation, such as ethanolic or heterolactic fermentation, the production of ethanol leads to
other fermented foods and beverages such as sauerkraut, kimchi, and wine.
The production of lactic acid forms the basic structure and texture of yogurt. However, other
molecules contribute to the taste of yogurt. These include acetaldehyde, an important flavor
substance in yogurt, and tyrosine, a product of proteolytic activity, but can cause bitterness when
the concentration is above 0.5 mg/ml

Fig 2

Enzymatic reaction in yogurt production.

Lactobacillus bulgaricus and Streptococcus thermophilus are not the only bacteria that can
convert lactose into lactic acid. Fresh milk begins acquiring microbes from the very moment it
leaves the cow. Milking equipment, people processing the milk, even bacteria in the air can all
contaminate milk. To prevent the milk from going bad, all milk sold at the grocery store is
pasteurized. Pasteurization is the process in which milk is heated for a certain length of time to
kill most of the microorganisms that might be present and to make it have a proper viscosity and
cooled to incubating temperature before inoculating the starter culture.. The U.S. Public Health
Service guidelines say that heating milk at 62.8°C (145°F) for 30 minutes or 71.7°C (161°F) for
15 seconds meets pasteurization standards. These standards are based on the amount of heat
necessary to kill most of the bacteria that are commonly found in milk. Of course, once you open
a container of milk, it can contain or acquire a mix of bacterial species that can ferment milk in
an undesirable fashion, as indicated in Table 1. Therefore, before you start making yogurt, it is
necessary to heat the milk so that the only bacteria it contains are the ones we will add! Adding
specific bacteria is called using a “starter culture”. Table 1 should be referred to when observing
your finished yogurt. In general, a “sniff” test is a good idea!
Below is a summary of the yogurt making process
 APPARATUS AND MATERIALS:

 500 mL glass beakers  Plastic cups

 Aluminum foil  Clean spoons
 Thermometer  milk (% fat does not matter)
 Hot plate stirrer  Phmeter

PROCEDURE

1. The group used one liter of milk. The milk was placed in a clean glass beaker. A
thermometer was then placed into the beaker.

2. Using a hot plate to the milk was heated slowly and indirectly until it reached 90°C,
Stirring was done to prevent the milk from burning onto the bottom of the beaker.

3. The milk was then taken off the hot plate and 40g of sugar was added and stirring was
done for about 10mins and the solution was set aside to cool. The beaker was covered
with aluminum foil to prevent contamination from the air.
4. The milk was cooled to 45°C in still air and then poured into a clean plastic cup.
5. 0.1g of culture was added and stirring was done thoroughly.

6. The cup was covered tightly with aluminum foil and placed into a 38-45°C incubator.

7. The yogurt was the fermented for 10 – 16 hours at 38-45°C.

8. An aliquot of the fermenting yogurt was picked every after one hour and PH was
determined using a Phmeter.

9. After the fermentation time, the yoghurt was stirred vigorously and flavors were added. It
was then packed and refrigerated.

RESULTS

TIME HOURS PH
1 6.60
2 5.89
3 4.76
4 4.51
5 4.49
6 4.43
 GRAPHY OF PH AGAINST TIME IN HOURS

4
PH
3

0
1 2 3 4 5 6

DISCUSSION
As seen from the graphy the pH of the solution has gone down with time adue to the fact that
lactose in the milk is constantly being converted to lactic acid which lowers the pH and continues
to do so as the bacteria continue growing. The production of lactic acid forms the basic structure
and texture of yogurt. However, other molecules contribute to the taste of yogurt. These include
acetaldehyde, an important flavor substance in yogurt, and tyrosine, a product of proteolytic
activity, but can cause bitterness. So mainly the product formed has a bitter taste but due to the
fact that we added sugar the taste becomes sweeter. As seen from the graph the specific pH level
was reached where most producers have a set point between 4 to 4.6 in which fermentation is
arrested by rapid cooling. The amount of lactic acid present at this pH level is ideal for yoghurt
giving it the characteristic tartness, aiding thickening and acting as a preservative against
undesired strain of bacteria. By verifying that fermentation continues to a predetermined pH
endpoint, yogurt producers can ensure their product remain consistent in terms of flavor, aroma
and texture. A deviation from the set pH can lead to a reduced shelf life of the yogurt or make
the product too bitter or tart. When comparing our made yogurt to the commercial yoghurt its
really hard to tell the difference but it could seen that the yogurt was just slightly less thick and
viscous that commercial yoghurt at around the same temperature.

SOURCES OF ERRORS

Some water could have been accidentally placed in the beaker at step 3 of the procedures hence
as a result some bacteria where introduced into the solution which could have interrupted the
growth of our desired bacteria.
 RECOMMENDATION

Do not eat the yogurt that has been for it was made in laboratory glassware that may have had
chemicals in it previously.

In procedure 2, the milk should not be allowed to boil.

In procedure 3, the thermometer should not be removed and one must careful not to get water in
the beaker.

Always wear an apron and goggles in the lab.

Use care handling the beaker of hot milk.

CONCLUSION

In this experiment yoghurt was produced by adding bacteria to milk and the product was
observed and compared with commercial yogurt hence I consider this practical a success.

REFERENCES

Food Microbiology (2008), Third Edition, Martin R. Adams and Maurice O. Moss, Publisher:
RSCPublishing, Pages: 323, 324 and 325.

Laboratory Manual: Concepts in Biology, (1997), 8th Edition, Eneger, Eldon D. and Frederick
C. Ross, Publisher: Wm. C. Brown Publishers, pages: 95-97.

Microbiology Concepts and Applications, (1993), second edition, Pelczar, Michael J., E.C.S.
Chan, and Krieg, Publisher: Noel R, McGraw Hill, pages: 847- 874.