1.

INTRODUCTION

Fermentation is a metabolic process that produces chemical changes in organic substrates

through the action of enzymes. In biochemistry, it is narrowly defined as the extraction of energy
from carbohydrates in the absence of oxygen. In food production, it may more broadly refer to
any process in which the activity of microorganisms brings about a desirable change to a
foodstuff or beverage. The science of fermentation is known as zymology.

Lactic Acid Fermentation

Lactic acid fermentation is a metabolic process by which glucose or other six-carbon sugars
(also, disaccharides of six-carbon sugars, e.g. sucrose or lactose) are converted into cellular
energy and the metabolite lactate, which is lactic acid in solution. It is an anaerobic fermentation
reaction that occurs in some bacteria and animal cells, such as muscle cells.

If oxygen is present in the cell, many organisms will bypass fermentation and undergo
cellular respiration; however, facultative anaerobic organisms will both ferment and undergo
respiration in the presence of oxygen. Sometimes even when oxygen is present and aerobic
metabolism is happening in the mitochondria, if pyruvate is building up faster than it can be
metabolized, the fermentation will happen anyway.

Acetic Acid Fermentation

Acetic acid also called ethanoic acid is organic compound. Acetic acid produced via
fermentation. Its pathway is conversion of glucose to ethanol and ethanol to acetic acid. In first
step, Saccaromyces cerevesiae (yeast) converts fermentable sugar of molasses into ethanol and
carbon dioxide. a process of oxidation in which alcohol is converted into acetic acid by the
agency of bacteria of the genus Acetobacter, especially aceti (as in the production of vinegar
from cider or wine)

1b) Outline The Factors Affecting Fermentation Process

 Temperature: The effect of temperature on microorganisms is numerous.

 pH: pH can affect the activity of enzymes and the charge status of cell membranes.

 Dissolved oxygen Concentration

 Foam Concentration

 Concentration of nutrients
2A) Characteristics Features Of Batch And Continuous Culture

Batch culture refers to a technique used to grow microorganisms in a limited supply of nutrients,
which declines when these are used up, or some other factor becomes limiting. Continuous
culture refers to a technique used for the production of microbes or microbial products in which
nutrients are continuously supplied to the fermenter.

2b) In A Tabular Form Differentiate Between Batch And Continuous Culture

Continuous culture Batch culture

Continuous culture is an open system. Batch culture is a closed system

Internal Environment The environment of the The internal environment of batch culture is
continuous culture is not changed during the changed with the progression of the
fermentation process. fermentation process.

Nutrients are continuously added to the Nutrients are added at the beginning of the
continuous culture throughout the process. process in batch culture.

Nutrients are not a limiting factor in a Nutrients become a limiting factor at some
continuous culture point in a batch culture

Lag and log phases are maintained in a Are three-phase such as Lag, log, and
continuous culture. stationary phases.

The process continues and the products are The whole process is stopped when the
continuously removed from the fermenter in a products are formed in a batch culture.
continuous culture.
2c) Enumerate the advantages of batch culture and continuous culture

The advantages of a batch culture are:

 Short duration.

 Less chance of contamination as no nutrients are added.

 Separation of batch material for traceability.

Disadvantage of batch culture

The major disadvantage of batch cultivation is its low productivity due to its high downtime
(non-production time which is used for cleaning, sterilization, and startup of another batch
cultivation) during two batch cultivations.

Advantage of continuous culture

 Continuous culture provides a better degree of control than a batch culture.

 Growth rates are regulated and maintained for extended periods.

 By varying the dilution rate, biomass concentration may be controlled.

 Secondary metabolite production may be sustained simultaneously together with growth.

Disadvantage of continuous culture

 Set up is more difficult,

 The maintenance of required growing conditions can be difficult to achieve.

 IF contamination occurs, huge volumes of product may be lost.

3) ANTIBIOTICS

Antibiotics are chemical substance produced by a living organism, generally a microorganism,

that is detrimental to other microorganisms. Antibiotics commonly are produced by soil
microorganisms and probably represent a means by which organisms in a complex environment,
such as soil, control the growth of competing microorganisms. Microorganisms that produce
antibiotics useful in preventing or treating disease include the bacteria and the fungi.
3a) ANTIBIOTIC PRODUCTION

Antibiotics are produced industrially by a process of fermentation, where the source

microorganism is grown in large containers (100,000 – 150,000 liters or more) containing a
liquid growth medium.

Oxygen concentration, temperature, pH, and nutrient levels must be optimal and are closely
monitored and adjusted if necessary. As antibiotics are secondary metabolites, the population
size must be controlled very carefully to ensure that maximum yield is obtained before the cells
die. Once the process is complete, the antibiotic must be extracted and purified to a crystalline
product. This is simpler to achieve if the antibiotic is soluble in organic solvent. Otherwise it
must first be removed by ion exchange, adsorption, or chemical precipitation.

Microorganisms used in fermentation are rarely identical to their counterparts in the wild. This is
because species are often genetically modified to yield the maximum amounts of antibiotics.
Mutation is often used and is encouraged by introducing mutagens such as ultraviolet radiation,
x-rays, or certain chemicals. Selection and further reproduction of the higher yielding strains
over many generations can raise yields by 20-fold or more. Another technique used to increase
yields is gene amplification, where copies of genes coding for enzymes involved in the antibiotic
production can be inserted back into a cell, via vectors such as plasmids. This process must be
closely linked with retesting of antibiotic production and effectiveness.

3b outline the importance antibiotic produce for industrial application

Antibiotics work in different ways to kill bacteria or stop them from growing. "All classes" of
antibiotics groups together In this portal, antibiotics are classified into one of the following
classes: penicillins, fluoroquinolones, cephalosporins, macrolides, beta-lactams with increased
activity (e.g. amoxicillin-clavulanate), tetracyclines, trimethoprim-sulfamethoxazole,
lincosamides (e.g. clindamycin), urinary anti-infectives, and other antibiotics. For more
information about antibiotic classes and example agents that fall into each antibiotic class,
explore Antibiotic Class Definitions.

3c
4a mention any 4 industrially importance amino acid

 Preservatives

Amino acids are also commonly used as preservatives in food and drink. Fruit juices are often
preserved with the use of cysteine as an antioxidant.

 Flavour enhancers

Several amino acids or their derivatives are used as flavour enhancers in foods. For example,
glycine and alanine are used to heighten flavour and taste. Another example is monosodium
glutamate (MSG), a derivative of glutamic acid, widely used in Asian dishes to intensify the
flavour. MSG produces a different type of flavour called ‘umami’ which is vital to many dishes.

 Amino acids in the chemical industry

Many amino acids are used as precursors for chemicals used in various industries, such as
pesticides and herbicides. For example, threonine can be used to produce herbicide azthreonam
and glycine can be used to produce glyphosate, another herbicide.

 Amino acids in the fitness industry

Several amino acids (leucine, valine, proline, alanine, cysteine, and isoleucine) are used in
supplements for muscle growth and body building. Building muscle involves ingesting protein
and amino acids that build up the proteins.

4b Briefly explain the microorganism responsible for the production of commercially

importance amino acid

Corynebacterium glutamicum is the workhorse of the production of proteinogenic amino acids

used in food and feed biotechnology. After more than 50 years of safe amino acid production, C.
glutamicum has recently also been engineered for the production of amino acid-derived
compounds, which find various applications, e.g., as synthons for the chemical industry in
several markets including the polymer market. The amino acid-derived compounds such as non-
proteinogenic ω-amino acids, α,ω-diamines, and cyclic or hydroxylated amino acids have similar
carbon backbones and functional groups as their amino acid [Link] for
metabolic engineering of C. glutamicum for production of amino acid-derived compounds will
be described, and where applicable, production from alternative carbon sources or use of genome
streamline will be referred to. The excellent large-scale fermentation experience with C.
glutamicum offers the possibility that these amino acid-derived speciality products may enter
large-volume markets.
4c) Describe The Method And Equipment For The Production, Harvesting And
Purification Of Amino Acid

Harvesting
The extraction from protein hydrolysates is not suitable for large-scale industrial production but
is relevant for specific amino acids, including l-cysteine, l-leucine, and l-tyrosine. Different
extraction processes can be developed based on the different chemical affinity and pH of the
amino acids for separation, For instance, l-cysteine may be obtained from keratin, which is found
in feathers, and can be extracted simply by using activated charcoal and concentrated
hydrochloric acid. Other amino acids, including l-leucine, l-alanine, and l-serine, can be
produced from waste material derived from animals

Most of these enzymes are obtained from microorganisms, such as Escherichia coli,
Aspergillus oryzae, and Pseudomonas sp. The main advantage of the enzymatic method is that it
can produce pure d-and l-amino acids in high concentrations with a very low formation of by-
products. The classical example is the enzymatic conversion of d,l-methionine after acetylation
to N-acetyl d,l-methionine, in which only the l-isomer is enzymatically converted by l-amino
acylase from Aspergillus oryzae to obtain l-methionine

Production And Purification

An amino acid can be produced in several ways such as by extraction from protein hydrolysate,
chemical synthesis, or biological processes (enzymatically catalysed synthesis and fermentation).
In particular, fermentation is emerging as the most promising due to the application of new
genetic engineering tools (Ikeda, 2003). Several enzymes have been used to catalyse the
production of desired amino acids such as hydrolytic enzymes, ammonia lyases, and NAD + -
dependent L-amino acid dehydrogenase (Pollegioni and Servi, 2012)

Aside from their benefits, enzymes are usually expensive and have limited stability, which are
the main drawbacks of enzymatically catalysed synthesis. However, fermentation is the most
economical and environmentally advantageous process to produce amino acids (Ikeda, 2003)

It can be done at mild operation conditions to prevent product degradation, and its maintenance
cost is low as compared to an extraction process (Ugimoto, 2010). The most common
microorganisms used to produce a broad spectrum of amino acids are Corynebacterium
glutamicum and Escherichia coli (Ikeda, 2003). High yields of lysine and glutamic acid (up to
50% w/w) have been obtained using genetically modified C. glutamicum which is also the
pioneer microorganism in producing essential amino acids, particularly L-lysine, L-valine, L-
isoleucine, L-threonine, L-aspartic acid, and Lalanine for commercial purposes (Ivanov et al.,
2013).

For example, Corynebacterium glutamicum and Brevibacterium spp. are two popular industrial
strains for the bulk production of amino acids [16]. Due to the free of endotoxin, C. glutamicum
is recognized as a safe strain and applied in the industrial production of L-glutamic acid and L-
lysine.

BCAAs also have positive effects on protein balance, liver regeneration, albumin
synthesis, physical and mental fatigue, and immune function [79]. Due to the increasing demand
of BCAAs in food market, the industrial production of BCAAs has reached 500 tons per year.

Methods and equipment

Glycine can be generated from serine under the catalysis of SHMT, so the accumulation of serine
precursor is favorable for the biosynthesis of glycine. However, chemical synthesis method is
preferred in the industrial production of glycine, which is the simplest achiral amino acid

Through these methods fermentation is one of the safe and developing processes for the
commercial production of amino acids because of the new genetic engineering applications that
are maximizing the yield and fertility of amino acids (Ikeda 2003). Essentially, amino acids exist
as L-enantiomers in all organisms.

4d) MECHANISM OF INSULIN PRODUCTION

Pancreatic β-cell dysfunction plays an important role in the pathogenesis of both type 1 and type
2 diabetes. Insulin, which is produced in β-cells, is a critical regulator of metabolism. Insulin is
synthesized as preproinsulin and processed to proinsulin. Proinsulin is then converted to insulin
and C-peptide and stored in secretary granules awaiting release on demand. Insulin synthesis is
regulated at both the transcriptional and translational level. The cis-acting sequences within the
5′ flanking region and trans-activators including paired box gene 6 (PAX6), pancreatic and
duodenal homeobox-1(PDX-1), MafA, and B-2/Neurogenic differentiation 1 (NeuroD1) regulate
insulin transcription, while the stability of preproinsulin mRNA and its untranslated regions
control protein translation. Insulin secretion involves a sequence of events in β-cells that lead to
fusion of secretory granules with the plasma membrane. Insulin is secreted primarily in response
to glucose, while other nutrients such as free fatty acids and amino acids can augment glucose-
induced insulin secretion. In addition, various hormones, such as melatonin, estrogen, leptin,
growth hormone, and glucagon like peptide-1 also regulate insulin secretion. Thus, the β-cell is a
metabolic hub in the body, connecting nutrient metabolism and the endocrine system. Although
an increase in intracellular [Ca2+] is the primary insulin secretary signal, cAMP signaling-
dependent mechanisms are also critical in the regulation of insulin secretion. This article reviews
current knowledge on how β-cells synthesize and secrete insulin.
5) Write an explanatory note on the production, application of the following

Bio-fertilizers

 Production

Bio-fertilizers are the product of fermentation process, containing specific individual or group of
soil microorganisms which improve the plant growth and productivity through supply of easily
utilizable form of nutrients. They are also known as bio-inoculants

There are three major ways of applying bio-fertilizers

1. Seed treatment

One packet of the inoculants is mixed with 200 ml of rice kanji to make a slurry. The seeds
required for an acre are mixed in the slurry so as to have a uniform coating of the inoculants over
the seeds and then shade dried for 30 minutes. The shade dried seeds should be sown within 24
hours. One packet of the inoculants (200 g) is sufficient to treat 10 kg of seeds.

2. Seedling root dip

This method is used for transplanted crops. Two packets of the inoculants is mixed in 40 liters of
water. The root portion of the seedlings required for an acre is dipped in the mixture for 5 to 10
minutes and then transplanted.

3. Main field application

Four packets of the inoculants is mixed with 20kgs of dried and powdered farm yard manure and
then broadcasted in one acre of main field just before transplanting.

Bio-pesticides production

Bio-pesticides are generally produced using synthetic and/or semi synthetic nutrient media to
attain increased yield and entomotoxicity level. The bio-pesticide production cost is higher than
that of chemical pesticides because of the use of synthetic medium to increase the yield of the
target products.

Application of bio pesticides

Bio-pesticides are in high agricultural demand due to their non toxic and eco-friendly
characteristics as compared to chemical or synthetic pesticides. The bio-pesticides are used as
microbial biological pest control agents who applied in farming processes to replace chemical
pesticides.
Bio fuels

Substances that produce heat and light energy on burning are called fuels. Some commonly used
fuels are wood, coal, cow dung cakes, kerosene, LPG, petrol, and diesel. We use fuels for
cooking, heating, in automobiles, and for the generation of electricity.

Applications of bio-fuels

Here are some of the application of bio-fuels

 Transportation.

 Energy Generation.

 Provide Heat

 Charging Electronics.

 Clean Oil Spills and Grease.

 Cooking.

 Lubricate.

 Remove paint and adhesive.

Waste degradation

Waste degradation also Biodegradation is the process by which organic substances are
decomposed by micro-organisms (mainly aerobic bacteria) into simpler substances such as
carbon dioxide, water and ammonia.16 Apr 2002

Production And Application Of Waste Degradation

In waste degradation processes, a chemical compound is transformed or eliminated by the

biological action of living organisms. In general terms, biodegradability is the tendency of a
lubricant to be ingested and metabolized by microorganisms.

The use of waste degradation for solving environmental contamination has primarily focused on
clean-up of sites contaminated with industrial bulk chemicals like oil products and (chlorinated)
solvents and on treatment of high-strength industrial
Sewage Treatment

Sewage treatment is a type of wastewater treatment which aims to remove contaminants from
sewage to produce an effluent that is suitable for discharge to the surrounding environment or an
intended reuse application, thereby preventing water pollution from raw sewage discharges.

Application of sewage treatment

The overall aim of treating sewage is to produce an effluent that can be discharged to the
environment while causing as little water pollution as possible, or to produce an effluent that can
be reused in a useful manner. This is achieved by removing contaminants from the sewage. It is a
form of waste management.

The major aim of wastewater treatment is to remove as much of the suspended solids as possible
before the remaining water, called effluent, is discharged back to the environment. As solid
material decays, it uses up oxygen, which is needed by the plants and animals living in the water.
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