GBE 302 BIOCHEMISTRY II LABORATORY

EXPERIMENT 1 Alcohol Fermentation

Submitted by Bra GM

Course Instructor: Asst. Prof. Dilek Telci Submitted to: Merve Erdem - Mge Yazc Experiment Date: 19.09.2011 Submission Date: 26.09.2011

Yeditepe University Istanbul

AIM: The aim of this laboratory is to learn about alcohol fermentaion, to determine the
carbondioxide formation and the enzymes that the yeast have or not.

THEORY
*Fermentation is an energy-yielding anaerobic metabolism that converts nutrients -such as sugars- into lactic acid, ethanol and acetic acid without oxidation.

*Alcohol fermentation is done by some kinds of bacteria and yeast. When glucose is present in the cell, by glycolysis, it is broken down into pyruvic acid. Then, by alcohol fermentation, this pyruvic acid is converted to carbon dioxide, ethanol and energy. C6H12O6 C2H5OH + 2CO2 H = -265 kcal/kg glucose

*By alcohol fermentation, bread, beer and wine can be produced and commonly Saccharomyces cerevisae is used.

*Acetic acid is produced by converting ethanol into acetaldehyde and then into acetate. CH3CH2OH CH3CHO CH3COO*Glucose, fructose and galactose are monosaccharides and they form disaccharides by joining. glucose + glucose maltose glucose + fructose sucrose glucose + galactose lactose

*For fermentation process, maltose, sucrose and lactose should be broken down into their monosaccharide units by enzymes.

*Glycerol is an alcohol and cannot be used in fermentation process.

*Calcium hydroxide is an inorganic compound and when reacts with carbondioxide, they form calcium carbonate as a white precipitate.

MATERIALS

Beakers Lab scale Pipettor Graduated cylinder Syringes (60 ml) Water bath Parafilm Water Glucose, lactose, sucrose and glycerol Yeast Calcium hydroxide

METHOD
0.6 g of glucose, lactose, sucrose and 1 ml of glycerol was put into 4 beakers. Then, 30 ml water was added on each beaker. When they dissolve, 0.6 g of yeast were added into each beaker. After having a homogeneous solution, they were transfered into syringes and the tips of the syrings were closed with parafilm. After 30 min incubation at 42-44C, the amount of gases formed was determined. To identify the gases, a few mililiters of 0.1 M calcium hydroxide was used. The gases were injected into tubes containing calcium hydroxide and shaked. Then, it was observed that if there was lime formation or not.

RESULTS

Glucose Amount of gas formed Lime formation 5 ml

Lactose -

Sucrose 16 ml

Glycerol -

Lime formation showed that the gas formed is carbondioxide.

DISCUSSION
In this experiment, alcohol fermentation by a yeast was observed. According to results, fermentation occurred only when glucose and sucrose used. For glucose; fermentation occurrence was obvious, because glucose is the main molecule that is used in glycolysis and so the alcohol fermentation. For sucrose; fermentation occurrence showed that the yeast used have the sucrase enzyme. So, the enzyme broke down the sucrose into fructose and glycose and fermentation occurred. For lactose; results showed that the yeast has no lactase enzyme, which can break down the lactose into galactose and glucose, and so it could not use the lactose for fermentation. Or, the yeast has lactase enzyme but it did not work in these conditions. For glycerol; there was no fermentation, because it was not a kind of molecule it is not a sugar- that can be used in glycolysis and fermentation. The amount of carbondioxide should have been more when glucose was used. This can be because the glucose is broken or the carbondioxide might be leaked.

REFERENCES
1. http://www.tempeh.info/fermentation 2. http://www.britannica.com/ 3. Lab manual