Nutritional Media Components
Nutritional Media Components
Nutritional Media Components
By,
Dr. Latika Shendre
Asst. Professor, DPU
COURSE DESCRIPTION:
Sr. No. Topic Description
1.
Introduction to Biochemical Historical background of
Engineering and Bioprocess Biochemical engineering,
Technology Introduction of industrially
important biotechnologically
products
2.
Isolation of microbes and Strain Isolation and preservation of
improvement industrially important microbes and
introduction of strain improvement
3. Design of fermentation media and Nutritional media components
inoculum development essential for growth of
microorganisms and product
formation, Media optimization
using conventional and statistical
designs, Inoculum development
for bacterial, fungal and yeast
strains
LEARNING OBJECTIVES
After this lecture, you will learn about:
Microbial nutrition and role of different nutrients in the growth of
microbes.
Growth medium and its various types.
Microbial growth and Phases of growth curve.
Effects of Different factors like temperature, pH etc. on the growth of
Microbes.
Methods used for quantification of growth.
MICROBIAL NUTRITION
• The purpose of microbial nutrition is to obtain energy and construct
new cellular components for growth.
Nutrient Requirement:
The major elements: these are also known as Macro Elements or
Macronutrients because these elements are required in high
amounts by the microbes. These includes C, H, O, N, S, and P.
The minor elements: these are also known as micro elements or
micronutrients as these are required in low amounts by the microbes.
These includes Ca, K, Mg, Fe.
• The trace elements: these elements are required in very low
amounts.
These are not essential elements for the growth of the microbes but
these are involved in biological functions.
E.g. zinc (Zn2+) is present at the active site of several enzymes.
6. It will cause minimal problems in the production process particularly aeration and
agitation, extraction, purification and waste treatment.
1. The Basic Nutrient Requirements of
Industrial Media
• All microbiological media, whether for industrial or for laboratory
purposes must satisfy the needs of the organism in terms of carbon,
nitrogen, minerals, growth factors, and water.
• In addition they must not contain materials which are inhibitory to
growth.
NUTRIENT REQUIREMENT
1. Carbon (C), Hydrogen (H) and Oxygen (O):
Each and every organism requires carbon, hydrogen, oxygen and
electrons for their growth and development.
These elements are main constituent of cellular material.
The requirement of C, H, and O can be satisfied together as most of
the carbon sources like glucose are often attached to hydrogen and
oxygen.
2. Nitrogen (N):
Required for the synthesis of amino acids, purines, pyrimidines,
enzyme cofactors and other substances.
3. Phosphorus (P):
Required for the synthesis of nucleic acids, phospholipids,
nucleotides, cofactors, some proteins and other cellular components.
4. Sulphur (S):
Required for the synthesis of cysteine, methionine, biotin, thiamine
and some carbohydrates.
5. Potassium salts (K):
It is cellular inorganic cation and cofactors for certain enzymes.
6. Magnesium (Mg):
It is cellular inorganic cation and cofactors for certain enzymes.
7. Calcium (Ca):
8. Iron (Fe):
It is the component of cytochromes and certain non-heme iron proteins and cofactors for
enzymatic reactions.
Carbon: Provides Nitrogen: Needed for the Minerals:
both energy and synthesis of amino acids,
Required for the
carbon units for lipids, enzyme cofactors and
other substances. activity in cell.
biosynthesis
A. Natural medium
B. Semi-synthetic medium
C. Synthetic medium
A. Natural medium:
culture media which consist only of naturally occurring biological
fluids e.g. Milk, urine, diluted blood, vegetable juices, meat extracts,
blood etc is known as Natural media.
B. Semi-synthetic medium:
culture media whose chemical components are partially known
and partially unknown are termed as Semi-synthetic culture media. e.g.
Potato dextrose agar (PDA), nutrient agar etc.
C. Synthetic medium:
culture media whose chemical composition is completely known
e.g. Mineral glucose medium, Richard’s solution etc. These media
are very useful in studying the physiology of microbes.
2. Based on application or function, media can be classified
as follows:
A. Selective media:
This media is used for growth of only selected microorganism.
It provide nutrients that only enhance the growth of particular microbe.
For example, if a microorganism is resistant to a certain antibiotic, such as
ampicillin or tetracycline, then that antibiotic can be added to the medium.
Only those microbes will grow in this media that were resistant to incorporated
antibiotic while microbes that are not resistant will die due to the antibiotic
added to the medium.
Example of selective media include MacConkey agar media which is used for
culture of Gram-negative bacteria.
B. Differential media or Indicator media:
• Differential distinguish one microorganism type from another,
growing on the same medium.
• This type of media uses the biochemical characteristics of a
microorganism growing in the presence of specific nutrients or
indicators (such as neutral red, phenol red, eosin y, or methylene
blue) added to the medium to visibly indicate the defining
characteristics of a microorganism. This type of media is used for the
detection and identification of microorganisms.
• Example of differential media include Blood agar media which
contains bovine heart blood that becomes transparent in the
presence of β- hemolytic organisms such as Streptococcus pyogenes
and Staphylococcus aureus.
C. Minimal media:
A minimal medium is one which supplies only the minimal nutritional
requirements for the growth of a microorganism. It just has enough
ingredients to support growth hence called as “minimal medium”.
1. Lag Phase
2. Log or Exponential phase
3. Stationary Phase or Plateau phase
4. Death phase
PHASES OF GROWTH CURVE
1. Lag Phase:
There is NO increase in cell number in this phase.
In this phase, cells adapt to a new environment.
There is NO change in number, but an increase in mass in this phase.
Length of the lag phase depends on characteristics of microbial
species also in part by the media conditions.
2. Log Phase or Exponential Phase
Although the net growth rate is zero during the stationary phase, but
cells are metabolically active and produce secondary metabolite.
4. Death Phase:
After the period of the stationary phase, the bacterial population
decreases due to the death of cells.
The death phase starts due to the death of cells.
The phase starts due to exhaustion of nutrients, accumulation of
toxic products and autolytic enzymes.
There is a decline in the variable count and not in the total count.
Fermentation medium:
manufacturing processes.
• Some nutrients are frequently added in substantial excess of that required, e.g.
P, K; however, others are often near limiting values, e.g. Zn, Cu.
buffering capacity.
Different constituents of medium:
• Water:
• It is the major component of the most fermentation media.
• Clean water with consistent composition is required for medium
preparation.
• Mineral content of the water is important in brewing during mashing
process.
• Hard water containing high CaSO4 concn are better for English Burton
bitters beers
• Similarly, high content of carbonate in water is better for darker beers.
• Nowadays, the water may be treated by deionization or other techniques
and salts added, or the pH adjusted, to favour different beers.
• Energy sources
• Light for autotrophs,
• Most industrial MO are chemo-organotrophs & common source of energy is carbon
source from carbohydrates, proteins , lipids.
• Factors influencing the choice of carbon source
• Rate at which carbon source is metabolized directly influence the formation of biomass
& formation of primary and secondary metabolite.
• The main product of fermentation process will determine the choice of carbon source.
• The purity of carbon source may affect the choice of substrate.
• The method of media preparation mainly sterilization may affect the suitability of
carbohydrates for individual fermentation process.
• Regional availability of carbon source (Pfizer use 10 different carbon source for
antibiotic production depending on the geographical location of production site.)
• The choice of substrate is often controlled by Government legislation and local laws –
in EEC beet sugar is encouraged rather cane sugar.
Commonly used carbon sources:
• Carbohydrates are the most commonly used as carbon source.
• Most widely available carbohydrate source is starch obtained from maize grain.
Other sources of starch are cereals, potatoes and cassava.
• Partially germinated barley grains when heated gives a material known as malt, it
contains variety of sugars besides starch.
• Sucrose is obtained from sugar cane and sugar beet. In fermentation media it is
commonly used in impure form known as molasses, which is the residues left after
the crystallization of sugar.
• Molasses can be used in the production of both:-
• Low value/ bulk products e.g., ethanol, SCP, organic and amino acids and some
microbial gums as well as high value/low bulk products e.g., antibiotics, enzymes,
vaccines and fine chemicals.
• In India cane molasses are mainly used in alcohol industry for the production of
sprit, country liquor or other liquor brands like rum, brandy, whisky.
• Cane molasses contains 21.2 % Invert sugar, 33.4 % sucrose and lacks raffinose,
whereas beet molasses contains 1% invert sugar, 1% raffinose and 48.5%sucrose.
• Oils and fats:
• Oils were first used as carriers for antifoams during antibiotic production.
• Vegetable oils such as olive, maize, cotton seed, linseed, soya bean are used as carbon
source because of their content of fatty acids, oleic, linoleic, linolenic acids.
• Oil contains much more energy approximately 2.4 times energy from glucose per
weight basis and thus oil also have volume advantage i.e., more energy requirement,
oils require less volume in fermenter as compared to glucose or sucrose.
•
• During fermentation if substrate purity is of upmost concern Glycerol trioleate is
used.
• Methyl oleate is used as the sole carbon substrate source in cephalosporin production.
• Hydrocarbon and their derivatives:
• Several n- alkanes have been used as carbon source for the production of :-
• Organic acids
• Vitamins and cofactors
• Nucleic acids
• Antibiotics
• Enzymes
• Ammonia is used for pH control and major source of nitrogen in defined medium for the
production of human serum albumin by Saccharomyces cerevisiae.
• Other proteinaceous nitrogen compounds which serve as source of amino acids are:-
• Corn steep liquor
• Soya meal
• Cotton seed meal
• Distiller solubles
• Peanut meal
• Corn steep Liquor (CSL): It is the byproduct left after the extraction of starch from maize.
• It is primarily used as a nitrogen source, also contain lactic acid, traces of reducing sugars
and complex polysaccharides (both C & N source).
• It is originally used for the production of penicillin. But now it is used in many fungal
antibiotic production medium.
• Nitrogen sources
• Soya meal: It is material left after deoiling of soya bean seeds. It contains about 8% w/w
Nitrogen. It has complex nitrogenous source than CSL. It is used in production medium
for Streptomycin.
• Cotton seed meal: It is clean, yellow, finely ground powder prepared from the embryo of
Cotton seed. It contains 56% protein, 24% carbohydrate, 5% oil and 5% ash. Ash basically
contains Fe, Ca, P, Cl and SO4. It is used in production medium of tetracycline.
• Distiller solubles: Residues (6-8% w/v total solids) left after the distillation of alcohol
from fermented grain or maize are further screened (removal of suspended solid residues),
leaving the effluent. Thereafter, effluent is further concentrated until solid reaches 35% w/v.
• To yield evaporator syrup, it is drum dried to yield Distiller solubles ,which contain
proteins.
• Minerals:
• These are very crucial for growth and metabolism of organism.
• Essential minerals are Ca, Mg, K, Cl, P & S. These are required in large conc (major
ingredients) as compared to other essential minerals like cobalt, copper, iron, manganese,
molybdenum and zinc (due to their presence as impurities in major ingredients do not
required in large amount).
• The yield of product formation varies linearly with the logarithmic concentration of key
metal.
• Theses are preformed factors added to the medium as some microorganism cannot
synthesize it.
• Vitamins are the most commonly required growth factor, besides vitamins
• Some amino acids, fatty acids or sterols are used as growth factors.
• Calcium panthothenate used in vinegar production,
• biotin in glutamic acid production.
• Some production strains may also require thiamine
• Many of the natural carbon and nitrogen sources used in media formulations contain
all or some of the required growth factors.
• Buffers:
• pH may be controlled by the external addition of ammonia, NaOH, and H 2 SO4. (acid or
alkali)
Besides the factors that regulate the growth of the cells, there are the
factors that regulate the product formation are incorporated in the media
formulations.
Precursor:
These are the molecules when incorporated into the medium they are
directly converted into the desired product.
When corn steep liqour is used in the production of penicillin the yield of
penicillin is increased from 20 unit /cm3 to 100 unit /cm3 as compared to
grown in other media.
Phenyl acetic acid is the most commonly used precursor for Penicillin
production.
Note: This service is not intended for secure transactions such as banking, social media, email, or purchasing. Use at your own risk. We assume no liability whatsoever for broken pages.
Alternative Proxies: