WHAT IS FERMENTATION??

FERMENTATION
the process of deriving energy from the oxidation of organic compounds, such as carbohydrates, using an endogenous electron acceptor, which is usually an organic compound. The term Fermentation is derived from the Latin verb "fevere" which means "to boil".

SUBSTRATES(molecule upon which an enzyme acts) :

Sugars are the most common. FERMENTATION PRODUCTS: Ethanol, lactic acid, hydrogen, butyric acid, acetone.
Yeast carries out fermentation in the production of ethanol in beers, wines and other alcoholic drinks, along with the production of large quantities of carbon dioxide. Fermentation occurs in mammalian muscle during periods of intense exercise where oxygen supply becomes limited, resulting in the creation of lactic acid.

HISTORY
French chemist Louis Pasteur was the first zymologist, when, in 1857, he connected yeast to fermentation. Pasteur originally defined fermentation as respiration without air. Pasteur performed careful research and concluded, "I am of the opinion that alcoholic fermentation never occurs without simultaneous organization, development and multiplication of cells.... If asked, in what consists the chemical act whereby the sugar is decomposed ... I am completely ignorant of it." German chemist and zymologist, Eduard Buchner, winner of the 1907 Nobel Prize in chemistry, later determined that fermentation is actually caused by a yeast secretion that he termed zymase.

ENERGY SOURCE IN ANAEROBIC CONDITIONS

Fermentation products contain chemical energy (they are not fully oxidized) but are considered waste products, since they cannot be metabolized further without the use of oxygen (or other more highly-oxidized electron acceptors). A consequence is that the production of adenosine triphosphate (ATP) by fermentation is less efficient than oxidative phosphorylation, whereby pyruvate is fully oxidized to carbon dioxide.

 Ethanol fermentation (performed by yeast and some types of bacteria) breaks the pyruvate down into ethanol and carbon dioxide. It is important in bread-making, brewing, ethanol fuel and wine-making.
**Usually only one of the products is desired; in bread-making, the alcohol is baked out, and, in alcohol production, the carbon dioxide is released into the atmosphere or used for carbonating the beverage. When the ferment has a high concentration of pectin, minute quantities of methanol can be produced.**

 is a biological process in which sugars such as glucose, fructose, and sucrose are converted into cellular energy and thereby produce ethanol and carbon dioxide as metabolic waste products. Because yeasts perform this process in the absence of oxygen, ethanol fermentation is classified as anaerobic.

A laboratory vessel being used for the fermentation of straw.

The chemical process of fermentation of glucose

One glucose molecule is converted into two ethanol molecules and two carbon dioxide molecules: C6H12O6 2C2H5OH + 2CO2 The process begins with a molecule of glucose being broken down by the process of glycolysis into pyruvate: C6H12O6 2 CH3COCOO + 2H+

This reaction is accompanied by the size difference of two molecules of NAD+ to NADH and a net of two ADP molecules converted to two ATP plus the two water molecules.

Pyruvate is then converted to acetaldehyde and carbon dioxide by an enzyme called pyruvate decarboxylase and requiring thiamine diphosphate as cofactor. The acetaldehyde is subsequently reduced to ethanol by the NADH from the previous glycolysis, which is returned to NAD+:

CH3COCOO + H+ CH3CHO + CO2 CH3CHO + NADH C2H5OH + NAD+

USES Ethanol fermentation is responsible for the rising of bread dough. Yeast

organisms consume sugars in the dough and produce ethanol and carbon dioxide as waste products. The carbon dioxide forms bubbles in the dough, expanding it into something of a foam. Nearly all the ethanol evaporates from the dough when the bread is baked. The production of all alcoholic beverages, except those produced by carbonic maceration, employs ethanol fermentation by yeast. Wines and brandies are produced by fermentation of the natural sugars present in fruits, especially grapes. Beers, ales, and whiskeys employ fermentation of grain starches that have been converted to sugar by the application of the enzyme, amylase, which is present in grain kernels that have been germinated. Amylase-treated grain or amylase-treated potatoes are fermented for the production of vodka. Fermentation of cane sugar is the first step in producing rum. In all cases, the fermentation must take place in a vessel that is arranged to allow carbon dioxide to escape, but that prevents outside air from coming in, as exposure to oxygen would prevent the formation of ethanol. Similar yeast fermentation of various carbohydrate products is used to produce much of the ethanol used for fuel.

 Lactic acid fermentation breaks down the pyruvate into lactic acid. It occurs in the muscles of animals when they need energy faster than the blood can supply oxygen. It also occurs in some kinds of bacteria (such as lactobacilli) and some fungi. It is this type of bacteria that converts lactose into lactic acid in yogurt, giving it its sour taste. These lactic acid bacteria can be classed as homofermentative, where the end product is mostly lactate, or heterofermentative, where some lactate is further metabolized and results in carbon dioxide, acetate or other metabolic products.

 Hydrogen gas is produced in many types of fermentation (mixed acid fermentation, butyric acid fermentation, caproate fermentation, butanol fermentation, glyoxylate fermentation), as a way to regenerate NAD+from NADH. Electrons are transferred to ferredoxin, which in turn is oxidized by hydrogenase, producing H2. Hydrogen gas is a substrate for methanogens and sulfate reducers, which keep the concentration of hydrogen sufficiently low to allow the production of such an energy-rich compound.

OUR EXPERIMENT
Anaerobic Respiration: ALCOHOLIC FERMENTATION

Objectives
To know the different factors that affect the process of anaerobic respiration To differentiate aerobic from anaerobic respiration

Saccharomyes cerevisiae
Yeast Aerobic and anaerobic respiration
TOOLS TO BE USED 1000 ml beaker 100 ml beaker 3 fermentation tubes Volumetric pipette Aspirator 10% glucose, 2% peptone, 0.5% yeast extract Live yeast suspension cotton

PROCEDURES
1. Fill three fermentation tubes up to the arm with nutrient medium ( 50 ml 10% glucose, 30 ml 2% peptone, 0.5% yeast extract). 2. Plug the mouth of fermentation tube and sterilize. (cool set-up). Describe the appearance of the medium.

3. Add live yeast suspension to fermentation tube 3. B. (observe after two days.) 4. . Add live yeast suspension to fermentation tube C. (observe after five days.)
5. Prepare and number two test tubes in rack. With a disposable Pasteur pipette. Transfer to tube 1 a pipetteful of fermentation from tube C. To tube 2, place a similar pipetteful of distilled water.

6. Add phenol red solution to both tubes. 7. To another set of three numbered test tubes. Add the following samples given in the book.
8. Add to each tube 1 ml strong I2 KL and 1.5ml 1.5 M NaOH. Mix the content of each test tubes and allow to stand for 510mins.

9. Mix the remaining medium in fermentation C, and, with a wire loop, transfer a loopful of it on a glass slide and place a cover slip. Examine under LPO and HPO. Make an enlarged sketch of the yeast cells.

THE LABORATORY APPARATUS

THE SOLUTIONS

GLUCOSE

PEPTONE

YEAST EXTRACT

GATHERING AND STERELIZING OF SOLUTIONS

THE THREE FERMENTATION TUBES WITH 10% GLUCOSE, 2% PEPTONE, 0.5% YEAST EXTRACT

TUBE C

TUBE B

TUBE A

DATA GATHERING AND RESULTS

1. Describe the appearance of the medium in fermentation tube A? Clearer solution than the other two solutions. no moist. yeasts subside. 2. How does the medium in tube B compare with that in tube A? Presence of moist. yeasts subside. The presence of clustered yeasts above the solution.

3. What has happened to the medium in the arm of fermentation tube C after five days? There are a lot of yeast present in the arm of the fermentation tube 4. After removing the plug off the mouth of tube C, what substance do you smell out of the odor? It is the yeast that we smell. 5. Result of procedure 6 to prove the presence of Carbon dioxide from tube C. The medium is acidic because the solution turned yellow when phenol red was added.

Tube B after two days

Presence of moist Clustered white particles
White substances subside

IODOFORM TEST
TUBE NUMBER 1 SAMPLE 2.5ml distilled water
1.25 ml distilled water & 1.25% ethanol 2.5 ml medium from fermentation tube C
IODOFORM RESULT (+/-)

**1 ml strong I2 KL and 1.5ml 1.5 M NaOH was added to these test tubes to determine if there is ethanol.

BASIC MEDIUM (red)

TUBE1 Fermentation medium

ACID MEDIUM (yellow)

TUBE2 Distilled water

** 1ml of phenol red is added to the following test tubes to know if it is an acid or basic medium.

7. Drawing of few yeast cells under HPO

CONCLUSION
We therefore conclude that We anaerobic respiration produces ethanol and carbon dioxide through the conversion of glucose to pyruvate. Fermentation medium in tube C is acidic when we dropped phenol red and positive in iodoform test thus giving us the conclusion that ethanol is very present in fermentation tube C.

GROUP 7 SABALLA SURTIDA SY TAN TORRENA TRIBIANA