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INTRODUCTION......................................................................................................................... 1
OBJECTIVE ................................................................................................................................. 4
SCOPE AND LIMITATION ....................................................................................................... 7
PRINCIPLE/THEORY .............................................................................................................. 10
EXPERIMENT ........................................................................................................................... 13
AIM: ............................................................................................................................................. 13
REQUIREMENT:....................................................................................................................... 13
PROCEDURE ............................................................................................................................. 16
OBSERVATION ......................................................................................................................... 19
RESULT....................................................................................................................................... 21
BIBLIOGRAPHY ....................................................................................................................... 22
ACKNOWLEDGEMENT
I would like to thank my teachers, Mr Kaushal Mishra and Mr Ajay Singh for guiding me through this project and
for their valuable inputs which provided me with a constant nudge for improvement.
It is imperative to thank our Principal, Mr Nitin Tiwari for providing me the opportunity to work on this project.
This project and reading-up on the same has provided me with an in depth understanding of the topic. It has nurtured
my scientific temperament and curiosity.
Signature
In this project, time taken for fermentation of various fruit / vegetable juices
had to be compared. Fermentation is one of the oldest methods of processing
food into a form that is suitable for preservation.
In fermentation technology, we stress in understanding the various process
in fermentor and how various intrinsic factors influence the fermentation
process. Fermentation technology being an industrial microbiology subject
are geared in producing maximum amount of high economical fermentation
products. The objective of this project is to compare the rates of
fermentation of different fruit and vegetable juices. The information gained
from this experiment may be used by wineries to determine which fruit juice
ferments best. But it is difficult to understand and control the fermentation
process as it involves various components such as effect of substrates,
products inhibition, conditions and complex microbial interactions.
Fermentation is affected by several factors including the temperature, salt
concentration, pH, oxygen availability and nutrient availability. The rate of
fermentation can be controlled by manipulating any of these factors.
Temperature
The fermentation process is not only complex but always in a state of flux.
Process, we are therefore in a situation to always be adaptive and reactive to
these changes so that throughout the fermentation process we are always
sustaining the conditions in a narrow window of optimal fermentation
conditions.
In order to help us do this we need to know fermentation kinetics. When we
talk about fermentation kinetics we are talking about fermentation models.
Kinetics and modellings are very useful to us as tools to make fermentation
predictions and enhancing our experimental designs to be more focused to
the specific problems such as the rate limiting steps or product inhibition.
The study of fermentation kinetics helps us by providing clear quantitative
data for us to understand the process and improve the process accordingly.
Peering into observation ports might be good advertising gimmick for
fermentation technology but do not really help much in understanding the
process or even to control and predict the fermentation outcome. Subjective
observations will rarely help in producing optimum fermentation process
and thus affect profitability studies and making decisions.
Its numbers that count!
A VERSATILE REACTION
Sucrose is hence first converted to glucose and fructose with the enzyme
invertase, while enzyme zymase converts glucose and fructose to ethyl
alcohol.
Invertase
Zymase
Zymase
C6H12O6 + C6H12O6 2C2H5OH + 2CO2
Glucose Fructose Ethanol
To test for the presence reducing sugars to the juice, a small amount of
Fehling’s solution is added and boiled in a water bath. During a water bath,
the solution progresses in the colors of blue (with no glucose present), green,
yellow, orange, red, and then brick red or brown (with high glucose present).
A colour change would signify and the presence of glucose.
Sucrose (table sugar) contains two sugars (fructose and glucose) joined by
their glycosidic bond in such a way as to prevent the glucose isomerizing to
aldehyde, or the fructose to alpha-hydroxy-ketone form. Sucrose is thus a
non-reducing sugar which does not react with Fehling’s solution.(Sucrose
indirectly produces a positive result with Benedict’s reagent if heated with
dilute hydrochloric acid prior to the test, although after this treatment it is no
longer sucrose.) The products of sucrose decomposition are glucose and
fructose, both of which can be detected by Fehling’s as described above.
Pasteur’s salt
The Pasteur’s salts in solution act as a buffer to any acids the yeast may
create. Since yeast only converts sugar (most likely sucrose or glucose) to
ethanol under anaerobic conditions, and it is unreasonable to assume that
there will be no oxygen present in the laboratory, some acetic acid is created
as a result. The Pasteur salts act as buffers to the acidity so that the proteins
in the yeast do not become denatured.
EXPERIMENT
Aim:
To compare the rates of fermentation of some fruit/vegetable juices and
determine the substance which has the highest rate of fermentation amongst
the various samples taken.
Requirement:
a. Chemical Requirement
Pasteur’s salts
Yeast
Fehling’s reagent
b. Apparatus Requirement
Conical flasks
Test tubes
Beaker
Bunsen burner, tripod stand and watch glass
PROCEDURE
1. 5.0 ml of apple juice was taken in a clean 250 ml conical flask and
diluted with 50 ml of distilled water.
5. Step 4 was repeated after every 10 minutes until the reaction mixture
stopped giving any red colour or precipitate.
6. This time taken, i.e. time taken for the completion of fermentation was
noted.
7. All the above steps were repeated by taking 5 ml each of grape juice,
black grape juice, sweet lime juice, orange juice and carrot juice.
Precautions:
80
70
60
TIME TAKEN
50
40
TIME TAKEN
30
20
10
0
TOMATO ORANGE CARROT SWEET LIME APPLE
JUICES
RESULT
The time taken for fermentation of carrot juice was well before the rest of
the juices, it’s recorded time being 30 minutes. This means that carrot juice
has the highest sucrose content from the various samples taken. After 50
minutes orange and tomato juices gave positive test for fermentation with
Fehling’s solution. For sweet lime juice time taken for fermentation was 60
minutes and for apple juice it was 70 minutes.
BIBLIOGRAPHY