FERMENTATION MEDIA

Fermentation is the process in which a substance is broken down into a

simpler substance by using microorganisms.
In industrial microbiology, fermentation is a process in which microorganisms
are grown on a large scale (by mass culture) to produce a product.
Fermentation Media
 nutrient-rich solutions or substrates used to support the growth and
metabolism of microorganisms in industrial fermentation processes.
 play a crucial role in providing essential nutrients required for microbial
growth and product formation.
Media Formulation
Most fermentations require:
 So liquid media, often referred to as broth.
 solid-substrate (in some)
 pilot-scale fermentations
In most industrial fermentation processes there are several stages where media
are required:

 several inoculums (starter, culture) propagation steps

 main production fermentation
The initial step in media formulation is the examination of the overall process
based on the stoichiometry for growth and product formation.
Carbon source + Nitrogen Source + Oxygen + Other requirements
Biomass+Products + CO₂ + H₂O + Heat

 where biomass or primary metabolites are the target products, the

objective is to provide a production medium that allows optimal growth of
the microorganism
 for secondary metabolite production, such as antibiotics, the objective is
to provide an initial period of cell growth, followed by conditions
optimized for secondary metabolite production
 most fermentations, except those involving solid substrates, require large
quantities of water in which the medium is formulated
General media requirements include:
 1. carbon source
2. sources of nitrogen, phosphorus and sulfur

CARBON SORCES NITROGEN SOURCES

 Molasses  Corn steep liquor
 Malt Extract  Yeast Extract
 Starch & Dextrins Sulphite  Peptone's
Waste Liquor Soya Bean Meal
 Cellulose
 Whey
 Alkanes & Alcohol
 Fats and Oils
3. Water
4. Mineral
5. Vitamins & Growth Factors
6. Precursors
7. Inducers and Elicitor
8. Inhibitors
9. Cell Permeability modifiers
10. Oxygen
11. Antifoams
12. Animal Cell Culture Media
13. Plant Cell Culture Media
14. Culture-maintenance media
 certain media nutrients or environmental conditions may affect not only
the physiology and biochemistry, but also the morphology of the
microorganism.
 media adopted also depend on the scale of the fermentation
 for small-scale laboratory fermentations, pure chemicals are often
used in well-defined media
 industrial-scale fermentations primarily use cost-effective complex
substrates, where many carbon and nitrogen sources are derived from
natural plant and animal materials

Main factors that affect the final choice of individual raw materials
1. cost and availability
2. ease of handling in solid or liquid forms, along with associated transport
and storage costs, e.g. requirements for temperature control
3. sterilization requirements and any potential denaturation problems
 4. formulation, mixing, complexing, and viscosity characteristics that may
influence agitation, aeration, and foaming during fermentation and
downstream processing stages.
5. concentration of target product attained, its rate of formation and yield
per gram of substrate utilized
6. the levels and range of impurities, and the potential for generating
further undesired products during the process
7. overall health and safety implications
CARBON SOURCES
A carbon source is required for all biosynthesis leading to reproduction,
product formation and cell maintenance

In most fermentations, it also serves as the energy source

Carbon requirements may be determined from the biomass yield

coefficient (Y), an index of the efficiency the conversion of a substrate
into cellular material.

Y carbon(g/g) = Biomass produces (g)

Carbon Substrate Utilized (g)

For examples:
 Saccharomyces cerevisiae grown on glucose has a biomass yield
coefficient of 0.56 g/g under aerobic conditions and 0.12 g/g under
anaerobic conditions.

Carbohydrates are traditional carbon and energy sources for microbial
fermentations, although other sources may be used, such as alcohols,
alkanes, and organic acids.

Animal fats and plant oils may also be incorporated into some media,
 often as supplements to the main carbon source.
Molasses
 pure glucose and sucrose are rarely used for industrial-scale
fermentations, primarily due to cost
 molasses, a by-product of cane and beet sugar production, is a cheaper
and more usual source of sucrose
 dark-colored viscous syrup containing 50–60% (w/v)
carbohydrates, primarily sucrose, with 2% (w/v) nitrogenous
substances, along with some vitamins and minerals
 overall composition varies depending on the plant source, location of the
crop, climatic conditions under which it was grown and the factory where
it was processed
 Hydrol molasses can also be used, a byproduct of maize starch
processing that primarily contains glucose
Malt Extract
 aqueous extracts of malted barley can be concentrated to form syrups
that are particularly useful carbon sources for the cultivation of
filamentous fungi, yeasts and actinomycetes
 contains about 90% carbohydrate (dry weight) and 20% hexoses
(glucose and small amounts of fructose), 55% disaccharides (mainly
maltose and traces of sucrose)
 contain a range of branched and unbranched dextrins (15–20%)
 also contain some vitamins and approximately 5% nitrogenous
substances, proteins, peptides, and amino acids
Starch & Dextrins
 polysaccharides that are not as readily utilized as monosaccharides and
disaccharides but can be directly metabolized by amylase-producing
microorganisms, particularly filamentous fungi
 extracellular enzyme of fungi hydrolyzes the substrate into a mixture of
glucose, maltose or maltotriose to produce sugar
 maize starch is most widely used, but it may also be obtained from other
cereal and root crops
 for a wider use in fermentation, starch is converted to sugar syrup,
which contains mainly glucose
 starch is gelatinized and then hydrolyzed by dilute acids of
amylolytic enzymes, often microbial glucoamylases that operate at
elevated temperatures
Sulfite waste Liquor
 a by-product in the manufacture of wood pulp-by the sulfite process
 sugar-containing wastes derived from the paper' pulping, which possess
mixture of hexoses (80%) and pentoses (20%) that are primarily used for
yeast cultivation
hexoses include glucose, mannose and galactose, whereas pentose
sugars are mostly xylose and arabinose
 usually, the liquor requires processing before use as it contains sulfur
dioxide
 low pH is adjusted with calcium hydroxide or calcium carbonate, and
these liquors are supplemented with sources of nitrogen and phosphorus
Cellulose
 predominantly found as lignocellulose in plant cell walls, which is
composed of three polymers; cellulose, hemicellulose and lignin
 lignocellulose is available from agricultural, forestry, industrial and
domestic wastes
 few microorganisms can utilize it directly, as it is difficult to hydrolyze
 mainly used in solid-substrate fermentations to produce various
mushrooms
 potentially a very valuable renewable source of fermentable sugars once
hydrolyzed, particularly in the bioconversion to ethanol for fuel use
Whey
 an aqueous by-product of the dairy industry
 expensive to store and transport so-lactose concentrates are often
prepared for later fermentation by evaporation of the whey, resulting in
the low removal of milk proteins for use as food supplements
 lactose is generally less useful as a fermentation feedstock than sucrose,
as it is metabolized by fewer organisms; for example, S. cerevisiae does
not ferment lactose
 this disaccharide was formerly extensively used in penicillin
fermentations and it is still employed for producing ethanol, single-cell
protein, lactic acid, xanthan gum, vitamin B12 and gibberellic acid
Alkanes & Alcohol
 n-Alkanes of chain length C10–C20 are readily metabolized by certain
microorganisms; mixtures are usually most suitable for microbial
fermentations
 methane is utilized as a carbon source by a few microorganisms.
METHANOL
 has a high percent carbon content and is relatively cheap although only
a limited number of organisms will metabolize it
 only low concentrations (0.1-1% w/v) are tolerated by microorganisms;
higher levels are toxic
 during the fermentation of methanol, the oxygen demand and heat of
fermentation are high
ETHANOL
 less toxic than methanol
 used as a sole or co-substrate by many microorganisms, but it is too
expensive for general use as a carbon source
 its biotransformation to acetic acid by acetic acid bacteria remains a
major fermentation process
Fats & Oils
 hard animal fats that are mostly composed of glycerides of palmitic and
stearic acids are rarely used in fermentations
 plant oils (primarily from cotton seed, linseed, maize, olive, palm,
rapeseed, and soya) and occasionally fish oil may be used as the primary
or supplementary carbon source, especially in antibiotic production
 plant oils are mostly composed of oleic and linoleic acids, but tinseed
and soya oil also have a substantial amount of linoleic acid
The oils contain more energy per unit weight than carbohydrates.

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