MCB 407 (INDUSRRIAL MICROBIOLOGY)

OPTIMIZATION OF FERMENTATIONS

FERMENTATION PROCESSES

INTRODUCTION
For several thousand years, fermentation has been a major way of
preserving food. Microbial growth, either of natural or inoculated population,
causes chemical and/or textural changes to form a product that can be stored for
extended periods. The fermentation process also is to create new, pleasing food
flavours and odours. The major fermentation is used in microbiology are the
lactic, propionic and ethanolic fermentations. These fermentations are carried
out with a wide range of cultures, many of which have not been characterized.
ALCOHOLIC BEVERAGES
Microbes play prominent role in the production of alcoholic beverages
such as beer, wine, vodka, brandy, whisky e.t.c. The production of these
beverages is discussed below:
(i) Beer production
The brewing process starts with the malting of the grain followed by
conversion of the malted grain to soluble extract which is fermented by yeast to
yield beer. The methods employed determine the quality and the type of beer
produced. Careful control must therefore be exercised at each stage of
production to produce a beer of acceptable standard.
Barely beer can be divided into two broad groups:
(1) Top-fermented beer (ales)
(2) Bottom-fermented beer (lager)
This distinction is based on whether the yeast remains at the top of the
brew (top-fermented beer) or sediment to the bottom (bottom-fermented beer)
at end of fermentation.
Brewing yeast strains are of two major types. The top-fermenting and the
bottom fermenting yeast. The top fermenting yeast remains uniformly
distributed in the fermenting wort and are carried to the top by the CO2 gas
generated during fermentation whereas bottom yeast settle to the bottom. Top
yeasts are used in the brewing of ales and bottom yeasts are used to make lager.
The bottom yeasts are usually given the species designation Saccharomyces
carlbergensis and the top yeast are called Saccharomyces cerevisiae.
Fermentation by top yeasts usually occurs at high temperature, 14 – 23oC
while fermentation by bottom yeast occurs at 6 – 12oC.
Fermentation is accomplished in a shorter period of time (5 – 7days) for
top fermentation while it takes 8 – 14days for bottom fermentation. The
brewing process, specifically production of barley beer involves the following
unit operations.
(i) Malting: - This is carried out to promote synthesis of hydrolytic enzymes
such as alpha amylase, endo-β gluconase and peptidases to solubulize the
endosperm wall of the grain and secure the enzymytic breakdown of soluble
component to low molecular weight.
The malting process thus provides sugar for the yeast from which it
obtains energy and amino acids for it’s growth. Malting involves 3 processes.
These are steeping, germination and kelming.
Steeping: Involves the soaking of the grains in water to a moisture level
of 42 – 46% for a period of up to 48 hours.
Germination: After soaking, the chatted grain is allowed to grow for 4 –
5 days. This process develop the endosperm enzyme which will modify the
starch, proteins and cell wall of the endosperm into useful extract (soluble
materials released from malt during mashing).
Kelming: This is drying of germinated grain at temperature between 50 –
90oC to arrest growth and enzyme activity. The kelming process also reduces
the moisture content of the grain to about 3 – 5%. It can thus be stirred, develop
flavour and characteristic, and colour forming potentials.
After kelming, the kelmed malt can then be grind in a mill in readiness
for marshing.

(2) Marshing:- This is the central process of brewing and consist of mixing
the malt with adjuncts at temperature which are optimum for the saccharomytic
and proteolytic enzymes present in the malt. The particular marshing procedure
employed is of optimus importance to the character of the resulting beer.
Marshing extracts, those materials from the malt and adjuncts which can
be solubilized under the particular conditions employed and the liquid resulting
from mashing is known as the wort. Marshing is influenced by various factors
such as temperature, time, concentration of malt, starch and protein.
The most importance objective of marshing is to produce fermentable
sugar largely through the amenolytic degradation of solubilized starch. Two
enzymes are mainly responsible for this a and β amylase. Hops give beer the
bitter taste.
There are 3 widely contrasting mashing processes.

(i) Infussion marshing: - This is characterized by the biochemical

conversion of the grist (malt) at a single temperature (65oC) in one vessel which
serves for extraction and filteration. The marsh is not boiled in this process.

(ii) Decoction marshing: - In this case, starch conversion starts at a lower

temperature which is eventually raised sometimes by the removal, boiling and
return of part of the marsh and any adjunct used.

(iii) The double-marsh method: - In this case, the starchy adjuncts are boiled
and added to the malt. The warm marsh is finally transferred to a lauter turn
where it is filtered.

(3) Fermentation: - The filtered-cooled wort (15oC) is pumped into

fermentation vessels which are often made of stainless steel. It is pitched with
yeast (0.2kg/hl on it’s way to the fermentation vessel). The fermentation
proceeds for the desired number of days depending on the type of beer, during
which some fermentable sugars are transformed to CO2, glycerol, acetate and
ethanol. Fermentation eventually cease due to exhaustion of nutrients and
inhibition by ethanol.
C6H10O6 2C2H5OH + 2CO2
(4) Lagering and ageing: - At the end of fermentation, the product is beer
and it has a harsh taste and referred to as ‘green beer’. In England, where ale or
top beer is consumed, the beer is not processed further but drawn at this stage. It
is however, primed to improve it’s taste and appearance. Priming is done by
small amount of sucrose, invert sugar or a mixture of cereal starch hydroxylase
and invests sugar caramel. Primary serves as substrate fro secondary
fermentation as well as beer sweetner.
In the case of lager beer, a fermented beer is transferred to maturation
tank and in the process, it is aerated and this facilitates a 2o fermentation.
Lagering occurs at 0 – 3oC for 2weeks to several months. The CO2 produced
during the 2o fermentation purges out dissolved O2, H2S and other unwanted
violates.
Moreover, the beer acquires additional properties which make it harsh
taste mellow. Diacetyl vicinyl diketones formed during yeast fermentation are
taken by yeast preventing formation of odours. All suspended and undissolved
solid gradually settle to the bottom of the tank and the beer is decanted free of
yeast cells and other solids. It is filtered either with filter mass or filter sleet to
remove its haze with the aid of diatomaceous earth filter. In some breweries,
ascorbic acid is added to prevent oxidation of some of the beer components.

(5) Bottling: - The filtered beer is pumped to the bottling hall for bottling.
Some breweries, the beer is carbonated to the extent of 0.45 – 0.50% by weight.
In some other breweries, no CO2 is added, it is the CO2 generated during
secondary fermentation that is used for carbonation.

(6) Pasteurization: - Filled and cannel bottles are pasteurized before

labeling.
WINE PRODUCTION
The term wine refers to the alcoholic beverage made from the juice of
variety of fruits by the fermentative action of selected yeast adapted to the
particular type of wine followed by the ageing process. The term is frequently
used to demote the alcoholic beverage made from grape fruit. The yeast used in
the fermentation of grape wine is Saccharomyces cerevisiae var. ellipsoideus
which have been selected over the years. There are two types of grape wine viz:
Red wine which are fermented with the skin (red, purple) which contain the
authocyanin pigment while white wine are produced from white grape or the
juice of other grapes fermented without their skin.

SPIRIT
They are distilled liquor obtained from fermentation of yeast of various
carbohydrates containing several raw materials e.g. sugarcane, grains and other
plant materials. Distilled liquor includes whiskey, from barley, malt water, rice
and oats. Bourbon from corn, gin from grain, malt flavoured with jumper
berries, rum from grains aquavit from grains flavoured with caraway seed,
vodka from grain. The manufacturing step in the spirit industry consists of raw
material preparation, yeast propagation, fermentation, and treatment of
fermented liquor and in some cases maturation/ageing.

BREAD BAKING
The use of yeast as a leavening agent in baking dates back to the very
early history of the Jews, Egyptians, Greeks and Romans. In modern baking
practice, pure cultures of selected strains of Saccharomyces cerevisiae are
mixed with bread dough to make about desire changes.
Desirable characteristics of S. cerevisiae selected from commercial
production of bakers yeast includes the ability to ferment sugar in the dough
vigorously and to grow rapidly. This is as well as other characteristics for which
the strain was selected should be relatively stable. The CO2 produced during the
fermentation is responsible for the leavening or rising of the dough. The quality
of the product depends on the proper selection of yeast and the incubation
conditions as well as on the choice of materials. The major unit operations
involved in baking are weighing, mixing, molding, fermentation, baking,
cooling and packing.

FERMENTED MILKS
The majority of fermented milk products rely on Lactic Acid Bacteria
(LAB), fermented milks were produced for thousands of years before Louis
Pasteur discovered lactic acid fermentation. LAB includes species belonging to
the genera lactobacillus, lactocococcus, leuconostoc and streptococcus.
These bacteria are low G + C gram-positives that tolerate acidic
conditions, are non sporing and are aero tolerant with a strictly fermentative
metabolism.

CHEESE PRODUCTION
This is one of the oldest human foods. Cheese is classified based on
texture or hardness as soft cheese (cottage, cream, brie) semi soft cheese
(muenster, limburger, blue) hard cheese (cheddar, Colby, swiss) or very hard
cheese (parmesan). All these result from a lactic acid fermentation of milk,
which results in coagulation of milk proteins and formation of a curd. Rennin,
an enzyme from calf stomach but now produced by genetically engineered
microorganisms can also be used to promote curd formation.
After the curd is formed, it is heated and pressed to remove the watery
part of the milk (called the whey), salted and then usually ripened. The cheese
curd can be packed for ripening with or without additional microorganisms. In
some cases, molds are used to further enhance cheese.

PROBIOTICS
Microorganisms such as Lactobacillus and Bifidobacterium are being
used in the rapidly developing area of probiotics, the benefit beyond basis
nutritive value. Probiotics is the oral administration of either living or substances
to promote health and growth, has the potential to re-establish the natural
balance and return the host normal health and nutrition. The possible health
benefits of the use of such microbial dietary adjurants include
immunomodulation, control of diarrhea, anticancer effect and possible
improvement of Crohn’s disease.
Acidophilus milk is produced by Lactobacillus acidophilus, L.
acidophilus may modify the microbial flora in the lower intestine thus
improving general health. It is often in use as a dietary adjunct, especially for
lactose intolerant persons.
Another interesting group used in milk fermentation are bifidobacteria.
The genus bifisobacterium contain irregular, nonsporing, gram-positive rods
that may be club-shaped or forked at the end. Bifidobacteria are thought to help
maintain the normal intestinal balance while improving lactose tolerance.
Bifidobacterium – amended fermented milk products, including yoghurt are
now available in various parts of the world.

MEAT AND FISH

Beside the fermentation of diary products, a variety of meat products
especially sausage can be ferment. Country-cured hams, summer sausage,
salami cervelat, fish sauces (processed by halophilic bacillus species) etc izushi
and katsuobushi Pediococcus cerevisiae and Lactobacillus plantarum are most
often involved in sausage fermentation. Izushi is based on the fermentation of
fresh fish, rice and vegetable by Lactobacillus sp; katsuobushi results from the
fermentation of tuna by Aspergillus glaulus. Both meat fermentation originated
from Japan.

OTHER FERMENTED FOODS

Many other plant products can be fermented and these include sufu, which is
produced by the fermentation of tofu, a chemically
coagulated soybean milk product. To carry out the fermentation, the tofu curd is
cut into small chuncks and dipped into a solution of salt and citric acid. After
the cubes are heated to pasteurize the surfaces, the fungi Actinimucor elagans
and some mucor species are added when a white mycelium develops, the cubes
now called pehtze are aged in salted rice wine.
This product has achieved the status of a delicacy in many parts of the
western world. Another popular product is tempeh, a soybean mash fermented
by Rhizopus.