INTRODUCTION:

Fermentation is a bio-chemical reaction where the substrate is converted

into desired product with the help of micro-organism which may be yeast,
Mould or bacteria and the enzyme present in the Micro-organism actually
acts in the reaction.

Reaction:

C6 H12 O6 ------> 2 C2 H5 OH + 2 CO2

(sugar) (alcohol)
The yeast Saccharomyces cerevisiae that is used for the culture for Alcohol
fermentation is having two kinds of enzymes, ones INVERTASE and other is
ZYMASE. Invertase converts sucrose to invert sugar and Zymase converts
invert sugar into Ethyl Alcohol.

Fermentation is a dynamic process involving a series of reactions. These are:

(i) Enzymes added after cooling continuous to convert the gelatinized

starch to dextrins and fermentable sugars.
(ii) The yeast metabolizes sugar to acetaldehydes and then to ethanol.
The cycle is known as Glycolysis.
The lactic acid bacteria produce small amounts of a variety of products,
some of which are volatile and contribute to the congeners in the distillate.

Molasses is transported in the cells via specific carrier proteins called

Permeases where it is hydrolysed into 2 molecules of glucose. The
Permeases are substrate and require metabolic energy for operation.
 C6H12O6 --> 2 C2H5OH +2 CO2 + 2 ATP + heat

Two moles of ATP (Adenosine Triphosphate) are also produced which are
used to supply energy for cell maintenance and growth. Theoretically
conversion of 1gm of glucose via fermentation yields 0.511grms of ethanol.
This theoretical value is never obtained due to carbohydrate utilization for
cell maintenance, growth and formation of small amounts of glycerols and
higher alcohols.

Fermentation efficiency also depends upon factors such as yeast strain and
environmental parameters. In practice efficiency is 60-80%. An increase in
temp within a certain range increases activity. The condition in Fermentation
vats are such obtained so as to avoid bacterial action. During a normal
Fermentation heat is produced from active yeast growth and metabolism
which causes a rise in temp. Then this temp rise can drastically affect yeast
metabolism and ethanol. An average upper limit temp for growth is around
40 C with an optimum temp of about 30C. Increased heat tolerances are
obtained with media containing oleic acid. The maximum concentration of
ethanol that yeast can produce depends upon the yeast strain used. In
general, yeast cell growth is inhibited around 10 to 12% wt/vol. ethanol
while 20% ethanol will terminate cellular metabolism.

Ethanol Fermentation With Yeast:

The organisms of primary interest to industrial operations in

fermentation of ethanol include Saccharomyces cerevisiae, S. uvarum,
Schizosaccharomyces pombe, and Kluyueromyces sp. Yeast, under
anaerobic conditions, metabolize glucose to ethanol primarily by way of the
Embden-Meyerhof pathway. The overall net reaction involves the production
of 2 moles each of ethanol, but the yield attained in practical fermentations
however does not usually exceed 90 95% of theorectical. This is partly due
to the requirement for some nutrient to be utilized in the synthesis of new
biomass and other cell maintenance related reactions.
A small concentration of oxygen must be provided to the fermenting
yeast as it is a necessary component in the biosynthesis of polyunsaturated
fats and lipids. Typical amounts of O2 maintained in the broth are 0.05
0.10 mm Hg oxygen tension.
The relative requirements for nutrients not utilized in ethanol
synthesis are in proportion to the major components of the yeast cell. These
include carbon oxygen, nitrogen and hydrogen. To leaser extent quantities of
phosphorus, sulfur, potassium, and magnesium must also be provided for
the synthesis of minor components. Minerals (i.e. Mn, Co, Cu, Zn) and
organic factors (amino acids, nucleic acids, and vitamins) are required in
trace amounts.

Yeast are highly susceptible to ethanol inhibition. Concentration of

1-2% (w/v) are sufficient to retard microbial growth and at 10% (w/v)
alcohol, the growth rate of the organism is nearly halted.