Genetics

Genetics is defined as the branch of biology that deals with the study of genetic variation,
genes, and heredity.
Genetic has applications in medicine, research, Food industry and agriculture sector and can
be used on a wide range of plants, animals and microorganisms.
Application of genetics
1. Genetically modified organism (GMO),
2. Transgenic animals,
3. Genetically modified crop
i. Golden rice,
ii. Herbicide resistant crops,
iii. Pesticide resistant crops etc.,
4. Genetically modified foods (GMF)
The possible benefits of genetic engineering include:
• More nutritious food
• Tastier food
• Disease- and drought-resistant plants that require fewer environmental resources (such as water
and fertilizer)
• Less use of pesticides
• Increased supply of food with reduced cost and longer shelf life
• Faster growing plants and animals
• Food with more desirable traits, such as potatoes that produce less of a cancer-causing
substance when fried
• Medicinal foods that could be used as vaccines or other medicines
Genetically modified organism (GMO)
What is a GMO?
A genetically modified organism (GMO) is an organism (plant, animal, bacteria or virus)
whose genetic makeup has been modified for a particular purpose. The organism does not occur
naturally in this modified state. For example, a plant can be modified to carry an additional
gene found in another living thing (such as a bacteria) to protect itself against insect pests.
Genes carry the information or the “recipe”, in the sequences and structures of DNA, which
gives the organism its specific characteristics. Genes can be added, removed or changed, using
modern biotechnology methods. Because genes are common to life on earth, genes can be
transferred from one organism to another and even between non-related species. This
manipulation can produce a product with new characteristics which may have advantages
The genetic modification of organisms or genetic engineering can be divided into the
following four main categories:
• Green Genetic Engineering (or agro-genetic engineering): Aims to develop genetically
modified plants in agriculture or the food sector.
• Red/Yellow Genetic Engineering: Utilised in medicine, diagnostics (genetic tests) and gene
therapy as well as development and production of drugs (insulin, vaccines).
• Grey/White Genetic Engineering: This is the production of enzymes or fine chemicals for
industrial use with the aid of genetically modified micro-organism (e.g. development of
products for enhanced washing performance).
• Genetically Modified Animals: Utilised for specific food production (e.g. dairy cows
modified to produce allergyfree milk).
How to make a transgenic crop
The processes involved in making a transgenic crop are the following:
1. Identification of an organism containing the desired gene. This can be from a plant, animal
or micro-organism.
2. Isolation of the desired gene from that organism.
3. The creation of a modified genetic sequence by the fusion of the desired gene, a promoter
sequence which controls the functioning of the gene and a marker gene e.g. a fluorescent
protein or an antibiotic resistance factor which allows the gene's presence to be detected even
when the target gene is not being actively expressed.
4. Multiplication of the recombinant sequence, usually in bacteria, to produce multiple copies.
5. Insertion of the copies of the desired gene into the organism to be modified, using either a
particle (gene) gun or a biological agent.
6. Selection of those organisms, which have successfully taken up the desired gene using a
selection test that recognises only those organisms, which have adopted the marker gene.
7. Multiplication of the modified plants.
How are GMOs useful to us?
Food:
GM (genetically modified) plants are already being cultivated as crops and consumed by
humans and animals. Using genetic engineering, new improved varieties of crops can be
produced more quickly than with conventional breeding methods. Crops can be modified to
have valuable characteristics such as tolerance to drought and herbicides, resistance to disease
and insects, as well as improved nutritional content.
Medicine:
• Insulin as a treatment for diabetes was the first commercial healthcare product produced by
GMOs. Bacteria were genetically modified to have a copy of the human insulin gene, and the
protein is synthesized by the bacteria.
• GMOs can produce other medicines such as growth hormone.
• GMOs are used in current vaccines such as Hepatitis B (produced by yeast), and new vaccines
are being developed using GMO technology. In the future, plants may even be engineered to
contain the vaccines so that we may be able to eat our vaccinations rather go for an injection.
• In the future, GMOs may be used for gene therapy to correct certain genetic conditions.
Textiles: GM cotton has been created to be resistant to insect attack to improve the yield of the
crop.
GM crops and food
As the world population continues to grow and more people need food, it becomes necessary
to increase food production. This can either be though the clearing of new land for agriculture,
or by increasing yield of the crops on the same amount or less land. The first generation of
GMO technology has been directed at reducing the risks of various threats to crop yields such
as insect attack and virus infection. Scientists have been adding genes to crop plants to give
benefits to farmers. Four GM crops are cultivated in South Africa: insect-resistant cotton (since
1997), insect-resistant maize (since 1998), herbicidetolerant cotton (since 2000) and
herbicidetolerant soybeans (since 2001). (Herbicides are chemical products used to destroy
weeds, but not the crop plants.) In 2004, it was estimated that GM crops accounted for 24% of
yellow maize, 10% of white maize, 50% of soybean and 85% of cotton in South Africa. South
Africa now ranks eighth of GM crop producing countries. Latest statistics from 2007 indicate
that 51% of yellow maize, 62% of white maize, 80% of soybean and 90% of cotton produced
were GM crops. Measurements of GM content in food products published in 2006 showed that
90% of soy products and 60% of maize products tested in South Africa contained GMOs.
Benefits
• Environmental benefits: Farmers can use less pesticide on insect-resistant GM plants. This
means fewer chemicals that can damage the environment are washed into the soil.
• Health benefits: Farmers need to use less pesticide on insect resistant plants, which results in
healthier food.
• GM crops are better protected by, and are not so susceptible to diseases, insects and
herbicides, allowing a more consistent yield.
• Pest resistant GM crops reduces the need to ‘blanket’ spray an entire crop, which impacts the
entire insect population. GM pest resistant crops effectively target only pest insects that eat the
crop.
• Higher yields of crops due to improved protection (it should be noted that in ideal conditions,
i.e. no pests or weeds, GM crops have no superior yield compared to their conventional
counterparts).
• It costs less to produce crops due to lower susceptibility to viruses and insects. The crops
have greater “hardiness”.
• Costs are potentially saved through a reduced need for pesticides and/or herbicides.
• GM crops can allow ‘no-tillage’ farming, which has benefits both for the farmer and for the
environment.
History of GMOs
10,000 Years Ago: Humans begin crop domestication using selective breeding.
1700s: Farmers and scientists begin cross-breeding plants within a species.
1940s and 1950s: Breeders and researchers seek out additional means to introduce genetic
variation into the gene pool of plants.
1980s: Researchers develop the more precise and controllable methods of genetic engineering
to create plants with desirable traits.
1990s: The first GMOs are introduced into the marketplace.
1995: Soy Beans
1995: Squash
1996: Corn
1996: Cotton
1997: Papaya
1999: Golden rice
2006: Alfalfa
2006: Sugar Beets
2013: Eggplant (Bangladesh only)
2016: Potatoes
2017: Apples
2018: Sugar Cane (Brazil)
2020: Pink Pineapple
 Additionally, with the production of golden rice, scientists have genetically modified food to
increase its nutrient value for the first time.
When it comes to genetic engineering of foods, there are different intended results by the
modification. Some crops, like rice, are modified to create a potential health benefit for its
consumers (especially in developing countries). Vitamin A is often added to the rice, creating
vitamin-rich "Golden Rice," which adds vitamin A into the diet of populations that consume
rice.
Transgenic animals
Transgenic technology has led to the development of fishes, live stock and other animals with
altered genetic profiles which are useful to mankind. Foreign genes are inserted into the germ
line of the animal, so it can be transmitted to the progeny. Transgenesis is the process by which
mixing up of genes takes place. Animal is one that carries a foreign gene that has been
deliberately inserted into its genome.
First transgenic other animals include pig, goat, cow, sheep, fish etc. Mouse – common
transgenic expt. The offspring was much larger than the parents. It was created by inserting a
human growth hormone gene in mouse genome. Animal was a ‘Supermouse’ created by Ralph
Brinster (U Pennsylvania) and Richard Palmiter (University of Washington) in 1982.
History
The first genetically modified organism was a bacterium created in 1973 by StanleyN. Cohen
and Herbert Boyer that contained genetic information from a variety ofdifferent species. The
first transgenic animals were mice created in 1974 by Rudolf Jaenisch, aprofessor of biology
at Massachusetts Institute of Technology. Then, in 1982, Ralph Brinster of the University of
Pennsylvania inserted thestructural gene for human growth hormone into mice embryos, and
noticed the mice withthe foreign gene grew much larger than those without the gene and also
passed this traitto its offspring.
Examples of transgenic animals
• Transgenic fish
• Transgenic chicken
• Transgenic mice
• Transgenic rabbits
• Transgenic goats
• Transgenic sheep
• Transgenic pigs
• Transgenic cows
• Transgenic dogs
• ANDI (transgenic monkey)

Genetically modified crop Golden rice

Rice is a staple food crop for over half of the world's population, supplying people in Asian
countries with 30–72 percent of energy intake, and being an important crop for targeted vitamin
deficiencies. With genetic modification technologies, the crops are updated with additional
nutritional values to enhance people's survival.
The new genetically modified rice (Oryza sativa) type is golden rice. It is produced for the
synthesis of vitamin A, to biosynthesize beta-carotene. It has a high content in vitamin A.
It was designed by Peter Beyer. Since rice grains (seeds) are yellow due to β-carotene, the rice
is commonly referred to as golden rice.
Golden Rice is a biofortified, genetically engineered crop. Biofortification increases the crop’s
nutritional value.
The Golden Rice Project was first launched in 1999, when two researchers, Ingo Potrykus and
Peter Beyer, proposed to the Rockefeller Foundation their proposal to genetically modify rice
in order to increase its nutrients.
-The International Rice Research Institute (IRRI) has agreed to grow Golden Rice to fulfill the
inventor's vision: to make the nutritional benefits of Golden Rice available as an alternative
vitamin A deficiency (VAD) intervention, at no additional cost compared to white rice, to
governments, small farmers or consumers in developing countries.
Golden rice is rice fortified by vitamin A, with a daffodil gene, and rich in carotene. Golden
rice differs from its parental strain by incorporating three genes for the biosynthesis of beta-
carotene. The parental strain in their leaves will naturally produce beta-carotene, where it is
involved in photosynthesis. The plant usually does not contain the pigment in the endosperm,
where photosynthesis does not occur.
Genetically modified herbicide resistant crops
Weeds, variously defined, have been in existence since time immemorial. They continually
interfere with human activity and crop production, despite the use of best weed management
practices. Prior to the advent of synthetic organic herbicides in the twentieth century, weeds
were controlled for thousands of years by mechanical, cultural, and biological means. These
methods are still widely practised in developing countries. The developed countries have seen
a rapid exodus of population from agriculture to pursue lucrative non agricultural occupations,
largely driven by mechanization and use of synthetic pesticides; this is also happening and
developing countries such as India. Discovered in the mid 1940s 2,4- dichorophenoxyacetic
acid was the first synthetic herbicide used for selective broad-leaves weed control. Since then,
several herbicides belonging to different chemical classes and possessing diverse modes of
action have been discovered and commercialized around the world. Herbicides have vastly
contributed towards increasing world food production in an efficient and economic, and
environmentally sustainable manner. The first such example is triazine-resistant canola that
was developed through a breeding programme in 1984. Thereafter, various methods of
resistance trait/germplasm selection such as microspore selection, seed mutagenesis, pollen
mutagenesis, tissue culture, cell selection, and transfer from a weedy relative have been used
for generating nontransgenic HRCs.
The initial efforts to develop transgenic HRCs using genetic engineering techniques resulted
in the release of bromoxynil-resistant cotton in 1995 and canola in 2000 (Table 2). However,
bromoxynil-resistant cotton and canola were discontinued because bromoxynil is not a broad-
spectrum herbicide and it failed to provide effective.

Future Herbicide-resistant Crop Technologies HRCs and other transgenic crops are here to
stay. While the benefits of these crops are plentiful, there are certain inherent consequences,
some obvious and a few not quite apparent. The most important issue requiring attention
following commercialization of HRCs has been the evolution of resistance to herbicide(s) in
weed populations. The agrochemical industry, seed companies, and related entities have
invested most of their resources in development of the next generation of HRCs (and other
transgenic crops) with the aim of diversifying the growers’ crop portfolio as well as combating
weed resistance by providing cropping technologies that allow application of more than one
mode of action herbicides.

Genetically modified pesticide resistant crops

Pest resistant GM crops (primarily cotton and maize), have been genetically modified so they
are toxic to certain insects. They are often called Bt crops because the introduced genes were
originally identified in a bacterial species called Bacillus thuringiensis. These bacteria produce
a group of toxins called Cry toxins.
Bt crops are grown widely in the USA, where an estimated 40% of GM maize is used in
industrial-scale biofuels (agrofuels) subsidised by the US government. The rest of this maize
is mostly used in animal feed, as is Bt maize grown in Brazil and Argentina. Some Bt maize is
also grown in South Africa.
Bt maize is also grown in small quantities in Europe, mainly in Spain, where it is used in animal
feed.
Bt cotton is the only GM crop authorised to be grown in India and China. It has also been
grown in smaller quantities in Pakistan, Colombia, Egypt and Burkina Faso.
Because the pesticide is produced inside Bt plants, rather than sprayed on the outside, it cannot
be washed off, so there are concerns that it may have adverse effects on humans, animals and
wildlife if and when the crop is eaten.
Bt crops are supposed to be grown with refuges of non-GM crops and plants to reduce the
likelihood of the targeted pests developing resistance (meaning that they are no longer killed
by the toxin produced in the plant). Despite this, resistant pests have been found in the US and
in India. In China, there have been reports of surges in other types of pest that are not effected
by the toxin produced by Bt cotton.
To deal with the problem of resistant pests, one experiment in the US used releases of pests
sterilised using radiation to try to reduce the population of pests in fields of Bt crops. The UK
company Oxitec is developing GM insects which it hopes to be paid to release in fields where
Bt crops are grown. The insects are genetically programmed to die before adulthood and are
supposed to reduce the population by mating with wild insects.
Genetically modified foods (GMF) and related regulations by standard
regulatory authorities
Most of the foods we eat today come from plants and animals that have been grown and bred
by humans for thousands of years. Traditionally, plants or animals with the most desirable
characteristics were chosen for food and then subjected to improvement through breeding the
next generation, leading to genetic changes over time. Modern biotechnology involving genetic
engineering techniques has provided new ways to allow selected individual genes to be
transferred from one organism into another, also between non related species. The resulting
organisms are referred to as genetically modified organisms (GMOs) i.e. organisms (plants,
animals or microorganisms) in which the genetic material has been altered in a way that does
not occur naturally by mating and/or natural recombination. GMOs are also referred to as
genetically engineered (GE) organisms, living modified organisms (LMOs) or transgenics.
Foods produced from or using GMOs are often referred to as genetically modified (GM) foods.
Examples include GE plants that are sold as food commodities (such as GM potato, GM maize,
GM papaya) and/or are further processed into food ingredients (such as GM canola for
modified oil composition).
In India, all GMOs including GE plants are regulated by the Ministry of Environment, Forest
and Climate Change (MoEFCC) as per rules notified under the Environment (Protection) Act,
1986. Guidelines and protocols are in place for safety assessment of GM foods. GM foods are
also subjected to regulations by the Food Safety and Standards Authority of India (FSSAI)
under the Food Safety and Standards Act, 2006.
In India, the regulation of all activities related to GMOs and products thereof are regulated as
per “Rules for the Manufacture/Use/Import/Export and Storage of Hazardous Microorganisms,
Genetically Engineered Organisms or Cells, 1989” (commonly referred to as Rules, 1989)
under the provisions of the Environment (Protection) Act, 1986. The Rules, 1989 are very
broad in scope, essentially covering entire spectrum of activities involving GMOs and products
thereof including sale, storage, exportation, importation, production, manufacturing,
packaging, etc. These rules are implemented by the MoEFCC, the Department of
Biotechnology (DBT), Ministry of Science & Technology, Government of India and State
Governments. Six competent authorities, their composition and roles have been notified under
the Rules, 1989.
Wine Production
• Wine is an alcoholic beverage produced through the partial or total fermentation of grapes.
• Other fruits and plants, such as berries, apples, cherries, dandelions, palm, Fig, Apricot and rice can also be
fermented.
• The species that are most widely used in wine production are Vitis labrusca and, especially, Vitis vinifera.
• Only a small part of the world is Wine producing. This is because the grape will only provide juice of the
quality required for conversion into consumable Wine where two climatic conditions prevail.
1. Sufficient sun-shine to ripen the grape
2. Winters that are moderate yet sufficiently cool to give the Wine a chance to rest and restore its strength for the
growing and fruiting season.

• Wines grapes contain all the necessary ingredients for wine, includingpulp, juice, and seeds that possess all
the acids, sugars, tannins, minerals,and vitamins that are found in wine.
• As a natural process, the frosty-looking skin of the grape, called "bloom," catches the airborne yeast that
ferment the juice of the grape into wine.
• The cultivation of wine grapes for the production of wine is called viticulture.
• Wine is characterized by color: white, pink or rose, and red, and it can range in alcohol content from 10
percent to 14 percent.
• Wine types can be divided into four broad categories:
– table wines: include a range of red, white, and rose
wines
– sparkling wines: include champagne and other "bubbly"
wines
– fortified wines: are table wines with brandy or other
alcohol added
– aromatic wines: contain fruits, plants, and flowers
• Red wine is extracted from the skin of red grapescontaining red
pigment (anthocyanin).
• During the preparation of red wine, all the anthocyanin
pigments are solubilized by the extract.
• Pink wine is obtained from either pink grapes or red grapes in
which fermentation last for only 12 to 36 hour and only less
amount of anthocyanin pigments are solubilized.
• White wine is prepared from the white grapes or from
the red grapes in which pigment involved in coloring
is removed.
• Wine production from crushed grapes is called
enology.
 Wine Making
• Wine making is something that has been done from thousands of years. Making wine is not just an art but
there is also a lot of science involved in the process.

• Smallest of mistakes in the process can have a major impact on the final product. Basically, wine is made in
5 different steps.
 Harvesting
• The moment at which the grapes are picked from the vineyard is what actuallydetermines the sweetness, flavour
and acidity of the wine.

• Fresh and fully ripened wine grapes are preferred as raw material for wine making.

• In cool climates, as in northern Europe and the eastern United States, however, lack of sufficient heat to produce
ripening may necessitateharvesting the grapes before they reach full maturity.

• The resulting sugar deficiency may be corrected by the direct addition of sugar or by the addition of a grape juice
concentrate.

• Because of the effect upon grape composition, proper timing of the harvest is of great importance.

• Premature harvesting results in thin, low-alcohol wines; very late harvesting may yield high-alcohol, low-acid
wines.

• Harvesting can be done by hand or by machines.

• Once picked grapes are taken to the winery and are then sorted in bunches.

• Under ripe and rotten grapes are removed.

 Crushing
• There are mechanical presses available that trod or stomp the grapes into ‘must’.

• Must is nothing but fresh grape juice which is the outcome of the crushingprocess and contains seeds,
solids and skins of the grapes.

• In modern mechanized wine production, the grapes are normally crushed andstemmed at the same time by a
crusher-stemmer.

• A roller-crusher may also be used.

• If white wine is being made then the seeds, solids and skins are quicklyseparated from the grape juice to prevent
the tannins and color for leachingin the wine.

• On the other hand, if red wine is being made, the seeds, solids and skins are allowed to stay in contact with the
juice to allow the juice to get additional tannins, flavour and colour.

• The drained pomace (crushed mass remaining after extraction of the juice from the grapes), from white or red
fermentations, may be used to provide distilling material for production of wine spirits.
 Fermentation
Pretreatments of Must

• Wines can be created by using the natural grape skin microorganisms.

• This natural mixture of bacteria and yeasts gives unpredictable fermentation results.

• To avoid such problems, one can treat the fresh must with a sulfur dioxide fumigant to killthe wild yeasts and
bacteria or sometimes pasteurized to destroy the natural microbiota.

• White musts are often turbid and cloudy, and settling is desirable to allow separation of thesuspended materials.

• Such measures as prior addition of sulfur dioxide and lowering of the temperature duringsettling help prevent
fermentation and allow the suspended material to settle normally.
• In many areas wineries centrifuge the white must to remove the solids.

• Musts are sometimes pasteurized, inactivating undesirable enzymes that cause browning.

• There is renewed interest in the prefermentation heat treatment of red musts to extract colour and deactivate
enzymes.
Fermentation

• Must now flow either into a stainless steel fermentation tank or a wooden vat (for finewines).

• Must then inoculate with a desired strain of Saccharomyces cerevisiae or S. ellipsoideus.

• Depending on alcohol tolerance of the yeast strain, final product may contain 10 to 18%alcohol.
 Primary Fermentation
• After 8-10 hours from inoculation with yeast the primary, alcoholic fermentation starts.

• This fermentation generally lasts for 8-10 days and during this period the yeast cells utilize the sugars in the
must and multiply, producing carbon dioxide gas and alcohol.

• The temperature used for red wines is typically 25° to 28 °C while for white wines it is 20° to 25 °C.

• During fermentation it is important to control the temperature and the oxygen concentration of the must.

• During alcoholic fermentation other substances (minor products): glycerol, acetic acid, higher alcohols, and
acetaldehyde are also produced.

• Wine quality is also defined by the quantity of these compounds and particularly by low concentrations of
acetic acid, higher alcohols and acetaldehyde.

• In the must used for red wine production, skins and seeds are present and during the alcoholic fermentation
the colour and tannin must be extracted.
 ...Primary Fermentation
• Seeds fall to the bottom of the tank while skins are pushed to the top of the tank by carbon dioxide (known
as "cap of pomace“).

• Grape skins are richer in flavour and colour (anthocyanin pigments) compounds and contact of the skin with
the must allows flavour and colour extraction.

• To extract the color and tannin in the skins, this cap must be broken and the fermenting must be thoroughly
mixed several times a day.

• When the sugar concentration of the must-have reaches about 10 g/L or less, usually in 7-10 days, the must
is drawn off the pomace.

• In some cases the wine may be allowed to remain with the skins and the seeds for seven or more days after
the fermentation are complete in order to obtain wines with a high tannin concentration.

• Pomace and wine are generally separated using either vertical or horizontal presses.

• The wine is then placed in a storage tank where the alcoholic fermentation runs to completion.

• This process requires about a week.

 ...Primary Fermentation
• A critical part of winemaking involves the choice of whether to produce a dry (no remaining free sugar) or a
sweeter (varying amounts of free sugar) wine.

• This can be controlled by regulating the initial must sugar concentration.

• With higher levels of sugar, alcohol will accumulate andinhibit the fermentation before the sugar can be
completely used, thus producing a sweeter wine.

• During final fermentation in the aging process, flavoring compounds accumulate and influence the bouquet
of the wine.
 Malolactic Fermentation

• When the sugar is fully utilized, the malolactic fermentation can take place.

• This process is used mainly for red wines but also for some white wines.

• In this process specific strains of the bacterium, Oenococcus oeni, convert malic acid to lactic acid.

• The fermentation reduces the titratable acidity and raises the pH which is advantageous in some wines.

• This fermentation is often initiated by inoculation with desired bacteria but can only be performed if the pH
is higher than 3.2 and the temperature is higher than 20 °C.

• Malolactic fermentation is performed for 2-4 week then the wine istransferred to other tanks for final wine
processing activities (fining,filtration, and aging).
 Clarification
• The purpose of fining is to remove excessive levels of certain winecomponents, to achieve clarity and to
make that clarity stable especially from a physicochemical viewpoint.

• Examples of such fining reactions are:

– The removal of tannic and/or brown polymeric phenols by protein-fining agents such as casein, albumin
or gelatin.

• Filtration in winemaking is a general operation that encompasses a wide range of conditions from the partial
removal of large suspended solids to the complete retention of microbes by perpendicular flow polymeric
membranes.

• Microbial growth during the fermentation process produces sediments, which are removed during racking.

• Racking can be carried out at the time the fermented wine is transferred to bottles or casks for aging or even
after the wine is placed in bottles.

• Generally all wines are subjected to fining and/or filtration processes but only some wines containing high
concentrations of ethanol and tannins are selected for aging.
 Aging and Bottling
• In general aging is used for red wines but some white wines (e.g.
Chardonnay) are also subjected to aging.

• Aging is a very complex process which has many effects on the wine.

• It can be performed in two stages, bulk and/or bottle.

• Bulk storage can be performed with inert containers like stainless steel or white oak barrels (like French
"barriques").

• In the latter case, however, the container reacts with wine; these reactionsare more intense when small barrels
are used.

• The most important reactions that occur during aging in an oak barrel are: wine oxidation, evaporation of
volatile components and reaction betweenwine and oak components.

Oxidation, in wine terms, refers to a group of chemical reactions that occur when wine comes into contact
with air. It can result in a wine losing its primary fruit characters and developing a brown hue.

• The aging period can range from few months to many years and is dependent on the wine type.

• At the end of this period wine is generally only filtered and bottled.

• Aging in the bottle ranges from some days to many years and is dependenton wine type.
INDIAN WINES REGIONS
Four major regions of vineyard: NASIK REGION, SANGALI REGION, BANGALORE REGION, HIMACHAL
REGION
Nasik Region
• Located in the state of Maharashtra.
• Region includes Pune, Nasik, and Ahmed Nagar.
• Above 800 meter from sea level.
• Several top wineries are located here.
• Chateau Indage, Sula wines.
Sangali Region
• Also located in Maharashtra.
• Nearly 750 meter above sea level.
• This region includes places like solapur, sangali, satara and latur.
Bangalore Region
• Nandi Hills located about around 45 kilometer North of Bangalore City.
• Grover Vineyards is located in Nandi Hills. It is above 800 meter sea level.
Himachal Region
• Located in northern India.
• It is upcoming state for the wine in India.
• Temperature varies from 20 C to 40 C.
• Unique Climate of this region attracts the wine makers to produce delicate wine grapes.

Indian Wine Law

The following particulars shall be marked legibly and indelibly on the label/crown/body of the container.
• Name and type of the material;
• Ethyl alcohol contents, in percent, by volume;
• Batch or code number;
• Month and year of Packing;
• Name and address of the producer
• Net volume in ml.
• Geographical region from where the grapes are produced
Organic acids

Organic acids are broadly distributed in nature, and humans have used them in their natural
forms since prehistoric times. Fermentation processes that involve the production of acetic acid
and lactic acid have been used in food preparation for centuries. Industrial production via the
microbiological route was started when the acids were identified as the main product in known
fermentation processes.Organic acids are traditional products of food technology and
biotechnology.

Organic acid is an organic compound that is characterized by weak acidic properties and does
not dissociate completely in the presence of water.

A few common examples include:

1. Lactic acid
2. Acetic acid
3. Formic acid
4. Citric acid
5. Oxalic acid
6. Uric acid
7. Malic acid
8. Tartaric acid
9. Butyric acid
10. Folic acid
One of the most important interventions for the control of microbial growth and the overall
safety of food products is the use of OAs and other chemical treatment for food preservation.

Organic acids such as benzoic acids, parabens, sorbic acid, propionic acid, acetic acid, lactic
acid, medium-size fatty acids, salts (sorbates, benzoates, propionates) and their mixture possess
strong antimicrobial activity and have been used as food preservatives, food contact materials
and sanitizers. and they have been declared as generally recognized as safe (GRAS).

Acetic acid is used as emulsifiers, stabilizers, preservatives, flavour enhancers and firming
agents. Formic acid which is the simplest carboxylic acid with one carbon atom, is used as a
preservative, acidifier in animal feed as well as a flavouring agent at low concentrations.
Propionic acid can be used as a pH control agent, preservative and flavour enhancer while
sorbic acid has found application as preservatives. Dicarboxylic acid includes adipic, fumaric
and succinic acid and have found application in beverage, feed and food preservation. Salts of
adipic acid such as calcium and magnesium adipate are used in food processes as sequestrants,
acidity regulators, baking additives, preservatives and flavour enhancers. Fumaric acid can be
used as pH control agents, flavour enhancers, firming agents and as emulsifiers.

Citric, lactic and malic acid have found application in beverage, food and animal nutrition.
Citric acid and its salts are used as sequestrants, pH regulators, preservatives, flavour
enhancers, and firming agents. Esters of citric acid can also be used as emulsifiers and solvents.
Lactic acid has been used for a long time as acidity regulators, preservatives, baking additives,
and flavor enhancers. Lactic acid can also be used as a humectant due to its hygroscopic
activity. Malic acid on its part can be used as synergists, acidity regulators, preservatives, and
flavour enhancers.

Vinegar Production

Vinegar is an acetic acid solution that is mild, aqueous and flavorful.

In the 6th century, the Babylonians were making & selling vinegar flavoured with fruit, honey,
malt etc. Egyptians records dates back to 300 BC.

Types of Vinegar

Type of Vinegar Raw material

Balsamic Vinegar Trebbiano Grape

Cane Vinegar Sugarcane

Champagne Vinegar Pinot Noir Grapes

Cider Vinegar Apples

Coconut Vinegar Coconut

Distilled Vinegar Grains

Malt Vinegar Barley & Grained Mash

Rice Vinegar Rice

Wine Vinegar White, Red or Rose Wine

Vinegar Production

Composition of Vinegar

Vinegar consists of an aqueous solution of acetic acid, water and a trace amount of other
chemicals. The concentration of acetic acid might vary. By volume, vinegar contains
approximately 5-8% of acetic acid.

Vinegar Production Flow Chart

Fruit extract + Yeast

↓
Alcohol Fermentation
↓
Acetic Acid Fermentation
↓
Ripening
↓
Filtration
↓
Pasteurisation (155° for 30 minutes)
↓
Bottling of Vinegar
(I) Alcohol Fermentation (30- 32⁰C)

There are several different ingredients that can be used to manufacture vinegar, such as grapes,
apples, oats, malted barley, sugar, beer, rice and other ingredients. However, vinegar was
probably originally created from wine as a commercial commodity.

Sugar is present in fruit juice or other liquid, and yeast enzymes use this sugar to produce
alcohol (ethanol) and carbon dioxide (CO2) gas.

(II) Acetic Acid Fermentation (27-37⁰C)

The two well-known acetic acid bacteria
are Gluconobacter and Acetobacter. Acetobacter bacteria are comparatively better acid
producers. By the action of the bacteria Acetobacter, the alcohol produced in the previous
process (ethanol) reacts with atmospheric oxygen to produce acetic acid and water.
Usually, the different flavours and aromas of vinegar are caused by the presence of organic
acids and esters that are derived from the fruit or other source material.
Filtration
Cross flow filtration involves mainly micro-porous hollow fiber membranes.

What is Single Cell Protein -SCP?

Single-cell protein refers to the crude; a refined or edible protein extracted from pure microbial
cultures, dead, or dried cell biomass. Single cell proteins are also called microbial proteins.
They can be used as a protein supplement for both humans and animals.

Microorganisms like algae, fungi, yeast, and bacteria have very high protein content in their
biomass. These microbes can be grown using inexpensive substrates like agricultural waste viz.
wood shavings, sawdust, corn cobs etc.

Sources of Single Cell Protein

A list of the microorganisms used for the production of Single Cell Protein is as follows:

Fungi (30-45%)

• Aspergillus fumigatus

• Aspergillus niger
• Rhizopus cyclopean

Yeast (45-55%)

• Saccharomyces cerevisiae

• Candida tropicalis
• Candida utilis

Algae (40-60 %)
 • Spirulina (spa)
• Chlorella pyrenoidosa

• Chondrus crispus
Bacteria (50-65%)

• Pseudomonas fluorescens
• Lactobacillus

• Bacillus megaterium

Production of Single-Cell Protein

The production is carried out in the following steps:
1. Selection of suitable strain.
2. Fermentation.

3. Harvesting.

4. Post-harvest treatment.

5. SCP processing for food.

1. Selection of suitable strain

The criteria used in choosing appropriate strains for SCP production includes, substrate must
be used as carbon and nitrogen source with high specific growth rates and productivity.
2. Fermentation
SCPs are produced generally by two types of fermentation.
Submerged fermentation•
Semisolid state fermentation•
Submerged fermentation is the one, where the substrate to be fermented is necessarily placed
always in a liquid that containing nutrients needed for the growth. The substrate is held in
fermentor that is operated continuously and simultaneously biomass product is continuously
harvested. The obtained product is further filtered/centrifuged and dried. This process has
higher operating cost. In semisolid fermentation, the substrate preparation is a simple solid
waste (Ex. Cassava waste).

3. Harvesting
When the colonies of microbes are fully developed, they are then harvested. The bulk of cells
are removed from the fermented by decantation.
4. Post harvested treatment
After harvesting the cells are subjected to a variety of processes. Post harvested treatments
include steps like separation by centrifugation, washing, drying.

Applications of Single-Cell Protein

1. Provides instant energy.

2. It is extremely good for healthy eyes and skin.
3. Provides the best protein supplemented food for undernourished children.
4. Serves as a good source of vitamins, amino acids, minerals, crude fibres, etc.
• Used in therapeutic and natural medicines for:
1. Controlling obesity.
2. Lowers blood sugar level in diabetic patients.
3. Reducing body weight, cholesterol and stress.
4. Prevents accumulation of cholesterol in the body.
• Used in Cosmetics products for:
1. Maintaining healthy hair.
2. Production of different herbal beauty products, like- Biolipstics, herbal face cream,
etc.
• Used in Poultry:
1. As it serves as an excellent and convenient source of proteins and other nutrients, it is
widely used for feeding cattle, birds, fishes etc.

Upstream and downstream processing

What is Bioprocessing?
 • Bioprocessing is defined as any process that uses complete living cells or their
components (e.g., bacteria, enzymes, chloroplasts) to obtain desired products.
• Bioprocessing is also essential to the production of renewable biofuels like ethanol and
biodiesel, therapeutic stem cells, gene therapy vectors, and new vaccines, among other
emerging industries and technologies.
• Bioprocessing includes two important processes - Upstream and downstream processes.
Upstream Bioprocessing
• The process of converting raw materials into a form that can be used in a biologic
manufacturing process is known as upstream bioprocessing.
• Harvesting and purifying natural products, culturing cells, and producing recombinant
proteins are all examples of this.
• Upstream bioprocessing's goal is to create a high-quality starting material for
downstream bioprocessing.
• The pre-fermentation stage
I. Isolation
II. Improvement
III. Producing of microorganisms
Downstream Bioprocessing
• The term "downstream bioprocessing" refers to the steps that take place after the initial
bioprocessing steps, which involve the production of a biological agent.
• There are three main steps involved in downstream processing- Purification,
Formulation, and Storage of the agent.
• Product recovery is another term for downstream bioprocessing. Bioprocesses produce
amino acids, antibiotics, organic acids, vitamins, and vaccines, among other
bioproducts. The bioproduct's desired quality is achieved during the downstream
process.
• Downstream processing is a technique used for the recovery and purification of such
as pharmaceutical or recombinant DNA products.

It consists of the following stages:

1. Solid-Liquid Separation- Whole cells and other insoluble substances are separated
from the fermentation broth. This is done using methods like flotation, filtration,
flocculation, centrifugation, gravity settling, etc.
2. Release of Intracellular Products- Several vitamins and enzymes present in the cells
are released in their active state for further purification and isolation. Other cells and
microbes are disrupted using physical, chemical, or enzymatic methods.
3. Concentration- The product that is to be extracted is a very small concentration. The
filtrate that is free of cells and other impurities still contains a large concentration of
water (80-98%) which needs to be removed to obtain the final product concentration.
Some of the methods used are evaporation, precipitation, liquid-liquid extraction,
adsorption, and membrane filtration.
4. Purification by Chromatography- Chromatography is used to separate biological
products having similar properties effectively. Some of the chromatographic
techniques include gel-filtration, ion-exchange, affinity chromatography, and
hydrophobic interaction.
5. Formulation- This step is necessary to maintain the activity of the biological
products during storage. For example, in the case of proteins, stabilizing agents such
as sugars, salts, and polymers are used. they are formulated as dry powders, solutions,
or suspensions.
What are Antibiotics?

Antibiotics are antimicrobial drugs obtained from other organisms (such as moulds, fungus and
some soil bacteria) to combat harmful microorganisms. However, they are not very useful
against certain microbes such as viruses.

Antibiotics are produced on a large scale by the fermentation process. This is a chemical
process which is induced by the microorganisms in a large tank. A large tank contains the
growth medium that provides nutrition for the microorganism’s growth. Optimization of
temperature, pH levels, oxygen and nutrient parameter in the fermentation tank is very crucial
to produce antibiotics. After the completion of antibiotics production, they are extracted and
purified through multiple physical processes. Finally, the purified antibiotics are converted into
crystal form.

Uses of Antibiotics

Today, the name antibiotic is synonymous with drugs that kill bacteria. And because of this,
mortality rates have reduced and lifespan has increased.

Antimicrobial therapy is used to destroy or prevent the microorganism’s growth. However, it

might not stop the damage that is already done. For example, the anthrax-causing bacterium
Bacillus anthracis does not actually kill the host; instead, it is the toxins that are produced that
do the deed. Antibiotics might kill the bacteria, but the toxins remain in the body and continue
to cause damage.

Today, two types of antimicrobial therapies are available:

1. Microbicidal therapy – It is used to kill microorganisms.

2. Microbiostatic therapy – It is used to prevent microorganism growth.

Types of antibiotics

There are hundreds of different types of antibiotics, but most of them can be classified into 6
groups.

Penicillin

Penicillin was first ever “true” antibiotic and it was discovered by a Scottish bacteriologist,
Alexander Fleming in 1928. It was widely used to treat the different types of bacterial infections
that plagued man. Ironically, Penicillin was an accidental discovery, but it was a landmark
discovery in the field of medicine. Penicillin is acquired from a mould called Penicillium
notatum (also known as Penicillium chrysogenum).

Penicillin widely used to treat a variety of infections, including skin infections, chest
infections and urinary tract infections.

There are different types of penicillin, which are grouped based on their effectiveness.

Penicillin VK and Penicillin G is natural penicillin that is produced directly from fermentation
of Penicillium crysogenum.

• Penicillin VK – It is used to fight against bacterial infections. Example: Ear infection.

• Penicillin G – More effective against gram-positive and gram-negative cocci bacterial

infections. Example: susceptible bacterial infections in the stomach.

• Cephalosporins (such as cefalexin) – used to treat a wide range of infections, but some
are also effective for treating more serious infections, such as sepsis (Sepsis is an
infection of the bloodstream that can cause newborn's immune system to attack its
body's own tissues and organs, such as the lungs, intestines or skin.) and meningitis
(Meningitis is the inflammation of the tissues surrounding the brain and spinal cord. It
is usually caused by infection).

• Aminoglycosides (such as gentamicin and tobramycin) – tend to only be used in

hospital to treat very serious illnesses such as sepsis, as they can cause serious side
effects, including hearing loss and kidney damage; they're usually given by injection,
but may be given as drops for some ear or eye infections
 • Tetracyclines (such as tetracycline, doxycycline and lymecycline) – can be used to
treat a wide range of infections, but are commonly used to treat acne and a skin
condition called rosacea

• Macrolides (such as azithromycin, erythromycin and clarithromycin) – can be

particularly useful for treating lung and chest infections, or as an alternative for people
with a penicillin allergy, or to treat penicillin-resistant strains of bacteria

• Fluoroquinolones (such as Ciprofloxacin, Quinolones and levofloxacin) – are broad-

spectrum antibiotics that were once used to treat a wide range of infections, especially
respiratory and urinary tract infections; these antibiotics are no longer used routinely
because of the risk of serious side effects.

Other antibiotics include chloramphenicol (used for eye and ear infections), fusidic acid (used
for skin and eye infections), and nitrofurantoin and trimethoprim (used for urinary tract
infections).
Production of Antibiotics:
Penicillin production involves following steps:
I) Preparation of Medium
II) Fermentation Process
III) Recovery Process

I) Preparation of medium
Carbon source: - Lactose, Glucose, Sucrose, Glycerol, Sorbitol.
Nitrogen source: - Ammonium sulphate, Ammonium acetate.

II) Fermentation
The fermentation conditions are:
Microorganism: Penecillium chrysogenum (Fungi)
pH: 6.5
Temperature: 26-28◦C
Types of fermentation: Aerobic (Fed batch fermentation)
Fermenter used: Stirred tank fermenter
 III) Recovery of Penicillin
A) Separation of mycelium
Mycelium is separated from the medium by employing rotary vacuum filter.
B) Extraction of Penicillin
Extraction of penicillin is carried out by employing the counter-current
extraction method. The liquid is immediately extracted with an organic solvent
such as amyl-acetate or butylacetate. The penicillin is then back extracted into
water from the organic solvent by adding enough potassium or sodium
hydroxide. The resulting aqueous solution is again acidified and re-extracted
with organic solvent.
C) Treatment of crude extract:
The resulted sodium penicillin is treated with charcoal to remove pyrogens (fever
causing substances). It is also, sometimes, sterilized to remove bacteria by using
Seitz filter. Then, the sodium penicillin is prepared in crystalline form by
crystallization.

History of Antibiotics

Alexander Fleming, a Scottish physician and microbiologist, discovered penicillin in 1928 at

St. Mary's Hospital in London. His discovery resulted in the development of antibiotics,
significantly reducing infection-related deaths.

Just before antibiotics were discovered, bacterial infections caused many deaths during the
First World War. Florey and Chain's work in 1938 enabled the industrial production of
antibiotics for medical use. In 1941, Selman Waksman coined the term "antibiotic" to
describe any small molecule a microbe produces that inhibits other microbes' growth.

Antibiotics provided much-needed relief as they helped cure mass infections after they were
mass-produced. They are now widely used to treat bacterial infections and save lives.
Antibiotic use has increased over the last century. Antibiotic production is naturally occurring.

Production of Antibiotics:
The Step-by-Step Manufacturing Process for Antibiotics

Antibiotic production can be divided into natural fermentation, semi-synthetic, and synthetic.
As more bacteria resist currently available antibiotics, research and development of new
antibiotics remain critical.

Antibiotics only treat bacteria-related infections. They do not work on viruses.

Antibiotics are produced in industries by fermenting the source microorganism in large

containers containing a liquid growth medium. They are derived from microorganisms or other
living systems.

Even though most antibiotics are naturally occurring, they are challenging to obtain in
sufficient quantities for industrial production. To accomplish this, it is necessary to isolate the
microorganism during fermentation. This microorganism is combined with culture for growth
in a large vessel. The antibiotic produced by this microorganism is harvested by filtering the
end product.

For a long time, antibiotics have transformed medicine by treating bacterial infections.

1. Start the Culture

Firstly, the microorganism used to produce the antibiotic is isolated and left to grow at a
suitable temperature. It multiplies many times before fermentation can begin.

In the lab, a starter culture is created from a previous sample that was cold-stored earlier. A
sample of the organism is placed on an agar plate. The initial culture is then grown in shake
flasks with food and other nutrients. This results in a suspension that can be moved to seed
tanks for continued growth.
Microorganisms are cultured in large seed tanks. Steel tanks are designed to provide an ideal
environment for microorganism growth.

Warm water and carbohydrate foods like lactose or glucose sugars are added to this seed tank
to help the specific microorganism survive, grow and thrive. Other carbon sources, such as
acetic acid, are added to the tanks, along with alcohol and hydrocarbons.

Nitrogen sources such as ammonia salts, vitamins, amino acids, and minor nutrients comprise
the seed tank's composition. Mixers in the seed tanks keep the growth medium moving, and a
pump delivers sterilised, filtered air.

The material in the seed tanks is transferred to the primary fermentation tanks after a few days.
Hygiene must be maintained, as contamination can spoil the culture and destroy the whole
batch.

2. Fermentation
The fermentation tank is a larger version of the steel seed tank, made of the same growth
medium. The seed tank provides a favourable environment for growth.
A seed tank is a large stainless steel kettle that provides an ideal environment for growing
bacteria.
Microorganisms are allowed to grow and multiply in this environment. During this process,
they excrete large amounts of the desired antibiotic.
The tanks are kept at the appropriate temperature. It is constantly agitated and fed a constant
flow of sterilised air and anti-foaming agents. Acids and bases are added to the tank because
pH control is critical for optimal growth. Fermentation is an essential step in the manufacturing
process.
3. Purification and isolation
Antibiotics are produced in abundance after three to four days. After this, the isolation process
can begin. The fermentation broth is purified by various methods depending on the antibiotics
produced.
Ion exchange could be used to purify water-soluble antibiotic compounds. These methods
separate the compound from the organic waste materials in the broth before passing it through
equipment that separates the other water-soluble compounds from the desired one.
A solvent extraction method is used to isolate an oil-soluble antibiotic like penicillin. In this
method, the broth is treated with organic solvents like butyl acetate or methyl isobutyl ketone,
which can specifically dissolve the antibiotic.
4. Refining
Antibiotic products can be sold as solutions in intravenous bags or syringes, such as pills, gel
capsules, or powders incorporated into topical ointments. After isolation, various refining steps
are taken depending on the antibiotic's final form.
The crystalline antibiotic is dissolved in a solution, placed in an intravenous bag, and then
hermetically sealed. The powdered antibiotic is physically filled into the bottom half of a gel
capsule before the top half is mechanically fitted on top.
Antibiotics are then transported to the final packaging station, stacked and placed in boxes here,
loaded onto trucks and delivered to distributors, hospitals, and pharmacies. The entire
fermentation, recovery, and processing can take five to eight days.
 Fermented Food and their biochemical and microbial analysis
Fermented Food
• Fermented foods are defined as “foods or beverages produced through controlled microbial
growth, and the conversion of food components through enzymatic action”. Many foods
have historically undergone fermentation, including meat and fish, dairy, vegetables,
soybeans, other legumes, cereals and fruits.
• Fermented foods are rich in beneficial probiotics and have been associated with a range of
health benefits from better digestion to stronger immunity.
• Fermentation is a natural process through which microorganisms like yeast and bacteria
convert carbohydrates such as starch and sugar into alcohol or acids.
• The alcohol or acids act as a natural preservative and give fermented foods a distinct zest
and tartness.
• Fermentation also promotes the growth of beneficial bacteria, known as probiotics.
• Probiotics have been shown to improve immune function as well as digestive and heart
health.
Health Benefits of Fermented Foods
Improves Digestive Health
Boosts Your Immune System
The bacteria that live in your gut have a significant impact on your immune system.
Makes Food Easier to Digest
Fermentation helps break down nutrients in food, making them easier to digest than their
unfermented counterparts.
Other Potential Benefits
• Mental health: A few studies have linked the probiotic strains Lactobacillus
helveticus and Bifidobacterium longum to a reduction in symptoms of anxiety and
depression.
• Weight loss: some studies have found links between certain probiotic strains
including Lactobacillus rhamnosus and Lactobacillus gasseri and weight loss and
decreased belly fat.
• Heart health: Fermented foods have been associated with a lower risk of heart disease.
Probiotics may also modestly reduce blood pressure and help lower total “bad” cholesterol.
Common Fermented Foods
There are many different types of fermented foods consumed around the world, including:
Traditional fermented foods
Traditional fermentation is a form of indigenous and spontaneous fermentation which involves
natural growth of microorganisms and their metabolites. Lactic acid bacteria and yeasts are the
major microorganisms involved in these spontaneous fermentations. However, in the
traditional condiments from alkaline fermentation,
Common traditional fermented foods
• Idli
• Sauerkraut
• Dosa
Idli:-
Idli is a very popular fermented breakfast fermented food of Indian origin made mainly of rice and
black gram. Traditionally, idli is steam cooked from the fermented batter of rice (Oryza sativa)
and dehulled black gram (Phaseolus mungo). Idli is white in color, round, soft and porous in
appearance with a mild sour taste. In terms of nutrition, idli offers a balanced amount of
carbohydrates, proteins along with essential amino acids and B-vitamins.

The principal components of idli include rice, black gram, salt and water. Idli preparation process
includes three major steps – soaking of rice and black gram, grounding and fermentation. The
standard idli batter preparation process consists of three major steps – soaking of rice and dehulled
black gram (4 – 6 hours), wet grounding and fermentation (12 – 18 hours at 30 ˚C).
Traditionally, idli preparation is as follows:
i) Rice and black gram were soaked separately,
ii) After draining the water, rice and black gram were grind separately with occasional
addition of water during grinding process,
iii) The rice and black gram batters were then mixed together with addition of a little salt,
iv) The mixture was allowed to ferment overnight at room temperature,
Tthe fermented batter was dispensed in special idli pans and allowed for steaming for
5–8 min. During fermentation of idli batter overnight the naturally occurring
microorganisms
Leuconostoc mesenteroides and Streptoccous thermophilus in grains/legumes/utensils
grow rapidly, outnumbering the initial contaminants and dominating the fermentation.
These microorganisms produce lactic acid and carbon dioxide that make the batter
anaerobic and leaven the product. Several aspects such as effect of raw materials, effect
 of fermentation or processing temperature and microorganisms involved in
biochemical and nutritive changes have been investigated
Sauerkraut:-
Sauerkraut is fermented shredded cabbage with a tangy, salty flavor. Sauerkraut fermented
white cabbage, a vegetable preparation important in the cooking of central Europe. Sauerkraut is
prepared by finely shredding white cabbage and layering the vegetable with salt in a large crock
or wooden tub. The cabbage is covered with a weighted lid and allowed to ferment, preferably at
below 15.5 °C for at least a month. Commercially made sauerkraut is canned or sold in bulk.
The antioxidant activity observed for sauerkraut in all studies was higher than that observed in raw
cabbage.
Requirements
Fresh cabbage, salt, weighing balance, knife, wooden vat or Jar
Procedure
1. Taken fresh cabbage
2. Cleaned, trimmed, and shredded into 2-5mm size.
3. Filled into wooden vats or cement tanks.
4. Salt is added at the rate of 2.25% and mixed thoroughly.
5. The top portion of the vat or tank is covered with plastic
6. Now enough weight is applied in order to make it compact and allow anaerobic
conditions to prevail for fermentation. When weight is applied, the salt dissolves in the sap
which is expressed by the pressure, and by osmosis it comes out from the cells.
7. After complete fermentation which is done for about 30 days or more until 1% lactic acid
is formed.
8. The Sauerkraut is removed from the vat and packed in cans, glass or plastic containers.
In cans, the fermented product is pasteurized at 74◦C for 3 minutes.
9. After that Sodium benzoate or potassium meta-bisulphite is added when the product is
packed unpasteurized. It is stored at + 5◦C.
Dosa:-
In its most simple form, dosa is crispy and crepe-like and is a very popular street food in
India. Dosa is famous for its simple ingredients and savory, slightly bitter flavor. It can be eaten
as a snack, breakfast, or anytime you’re in the mood for a delicious, savory meal. Originating in
southern India, the crispy dosa has been around since the 5th century.
I n g r ed i en t s
Rice flour and black gram flour, salt, water.

Fig. Flow chart preparation of dosa

Soy fermented foods and other foods
• Cheese
• Yogurt
• Miso
• Tempeh
• Soya Sauce
• Beer
• Wine
Cheese
Cheese is made by coagulation or curdling the milk with acid or rennin or both, drawing off the
whey and processing the curd.
Cheese is one of the by-products of the milk. It is processed form either cream of milk or cream
and milk mixed together
Types of cheese

• Cheese is a standard accompaniment to popular foods like burgers, pizza, Mexican

dishes, salad, and sandwiches.

• Alone, it can be a snack or an appetizer. It can be added to sauces, soups, pastries, and
many other dishes.

• There are thousands of varieties of cheese, ranging from mild to mature in flavor, and
low- to high-fat in composition. It can be made from the milk of cows, sheep, goats,
and other animals.

• Whole-milk cheese contains between 6 and 10 grams (g) of fat per 1-ounce (28 g),
serving. Of this, 4 g to 6 g is saturated fat.

• Low-fat or reduced-fat cheese is made with 2 percent milk. Non-fat cheese is made
with 0 percent or skim milk.
 • Fresh cheeses are cheeses that have not been aged, or matured. They usually have a
higher moisture content, softer texture, and milder taste than aged cheeses. Examples
include ricotta, cream cheese, cottage cheese, and mascarpone.

• Aged or mature cheeses are firmer in texture and tend to be aged for 6 months or
longer. The longer the aging process, the more concentrated or sharp the flavor.
Cheddar, Swiss and Parmesan are examples of aged cheeses.

• Processed cheese, such as cheese spread, American cheese, “cheese food” and “cheese
flavored” products cannot be categorized as cheese, and the label must reflect this.
These are shelf-stable products containing added ingredients such as flavor enhancers
and emulsifiers.

• Non-dairy cheeses, such as soy cheese and dairy, are suitable for people who do not
consume dairy products, but they are highly processed.

Health benefits
Dairy products are one of the best dietary sources of calcium and other nutrients. Here are eight
ways in which cheese might be healthful.
1. Bone health
The calcium, protein, magnesium, zinc, and vitamins A, D, and K content of cheese mean it
can contribute Trusted Source to healthy bone development in children and young adults.
2. Dental health
Cheese can enhance dental health. Calcium plays an important role in tooth formation, and cheese
is a good source of calcium.
3. Blood pressure
Statistics show that people who eat more cheese have lower blood pressure, despite some cheeses
being rich in fat and sodium.
• Calcium can help reduce blood pressure. Low-fat, low-sodium cheeses are recommended.
• A suitable low-sodium cheese would be low-fat or reduced-fat natural Swiss cheese.
• Other low-fat cheeses include cottage cheese, ricotta cheese, parmesan, feta, and goat’s
cheese, and low-fat cream cheese.
• Many cheeses are now available in “lite” versions, such as cheddar, brie, havarti, and feta.
 • Due to extreme processing, fat-free cheeses are not recommended as a regular part of the
diet, even for those looking to reduce calories or fat.
4. Healthy blood vessels
• Some cheeses contain high levels of cholesterol and sodium, suggesting that they can lead
to cardiovascular problems.
• However, in 2014, researchers concluded that dairy products could be a good source of
the antioxidant, glutathione. This antioxidant is crucial for brain health and for preventing
age-related neurodegeneration.
6. A Healthy weight
• Studies show that a person with a high body mass index (BMI) is more likely Trusted
Source to have low levels of calcium. Since cheese is a good source of calcium, there may
be benefits for people on a weight-loss diet.
7. Omega-3 fatty acids
• These have been found in some types of cheese, and especially those made from milk
produced by cows that eat Alpine grasses. Omega-3 fatty acids are believed to benefit
Trusted Source the cardiovascular system and the brain.
8. Healthy cells
• Cells need protein for building and repair. One ounce of cheddar cheese can offer 7 g of
protein. The amount of protein recommended for each person depends on their age, size,
and activity level.
General Cheese Processing Steps
• Standardize Milk
• Pasteurize/Heat Treat Milk
• Cool Milk
• Inoculate with Starter & Non-Starter Bacteria and Ripen
• Add Rennet and Form Curd
• Cut Curd and Heat
• Drain Whey
• Texture Curd
• Dry Salt or Brine
• Form Cheese into Blocks
• Store and Age
• Package
The times, temperatures, and target pH values used for cheddar cheese will depend on individual
formulations and the intended end use of the cheese. These conditions can be adjusted to optimize
the properties of Cheddar cheese for shredding, melting, or for cheese that is meant to be aged for
several years.
1. Standardize Milk
Milk is often standardized before cheese making to optimize the protein to fat ratio to make a good
quality cheese with a high yield
2. Pasteurize/Heat Treat Milk
Depending on the desired cheese, the milk may be pasteurized or mildly heat-treated to reduce the
number of spoilage organisms and improve the environment for the starter cultures to grow. Some
varieties of milk are made from raw milk so they are not pasteurized or heat-treated. Raw milk
cheeses must be aged for at least 60 days to reduce the possibility of exposure to disease causing
microorganisms (pathogens) that may be present in the milk.
3. Cool Milk
Milk is cooled after pasteurization or heat treatment to 90°F (32°C) to bring it to the temperature
needed for the starter bacteria to grow. If raw milk is used the milk must be heated to 90°F (32°C).
4. Inoculate with Starter & Non-Starter Bacteria and Ripen
The starter cultures and any non-starter adjunct bacteria are added to the milk and held at 90°F
(32°C) for 30 minutes to ripen. The ripening step allows the bacteria to grow and begin
fermentation, which lowers the pH and develops the flavor of the cheese.
5. Add Rennet and Form Curd
The rennet is the enzyme that acts on the milk proteins to form the curd. After the rennet is added,
the curd is not disturbed for approximately 30 minutes so a firm coagulum forms.
6. Cut Curd and Heat
The curd is allowed to ferment until it reaches pH 6.4. The curd is then cut with cheese knives into
small pieces and heated to 100°F (38°C). The heating step helps to separate the whey from the
curd.
7. Drain whey
The whey is drained from the vat and the curd forms a mat.
8. Texture curd
The curd mats are cut into sections and piled on top of each other and flipped periodically. This
step is called cheddaring. Cheddaring helps to expel more whey, allows the fermentation to
continue until a pH of 5.1 to 5.5 is reached, and allows the mats to "knit" together and form a
tighter matted structure. The curd mats are then milled (cut) into smaller pieces.
9. Dry Salt or Brine
For cheddar cheese, the smaller, milled curd pieces are put back in the vat and salted by sprinkling
dry salt on the curd and mixing in the salt. In some cheese varieties, such as mozzarella, the curd
is formed into loaves and then the loaves are placed in a brine (salt water solution).
10. Form Cheese into Blocks
The salted curd pieces are placed in cheese hoops and pressed into blocks to form the cheese.
11. Store and Age
The cheese is stored in coolers until the desired age is reached. Depending on the variety, cheese
can be aged from several months to several years.
12. Package
Cheese may be cut and packaged into blocks or it may be waxed.

Biochemical and microbial analysis of fermented food products

Biochemical analysis of fermented food products
Biochemical analysis determine by approve methods of AACC applied to determine pH and total
titratable acidity then determine moisture content, protein content by a Kjeldahl method, total fat
content ,ash content, reducing sugar, alcohol and starch content.
Microbial analysis of fermented food products
Microbial analyses of fermented food products analysis determine by using approve methods.
There is the some microbial analysis of fermented food products i.e. Aerobic Plate Count, Yeast
& Mold, Coliforms, Lactic Acid Bacteria, Spore-forming Mesophilic Aerobic Bacteria, Anaerobic
Plate Count, Total plate count.
Upstream and downstream processing
The bioprocess can be explained as a particular process that utilizes complete living cells or
some of their cellular components to get wanted items. The process is used in the creation of
nourishments, pharmaceuticals, flavors, energy, and synthetic concoctions with the guidance
of a biocatalyst. A catalyst in bioprocessing can be microorganisms, plants, or creature cells
in a bioreactor.
Alongside, it includes hereditary designing for the control of animals, plants, and
microorganisms. Downstream handling is required to expel debasements, mass volume
decrease, and concurrent convergence of the ideal item from the bioreactor.
Protein recuperation is touchy to working conditions because their capacity relies upon the
honesty of the sensitive 3D tertiary structure.
What is Bioprocessing?
• Any process that uses complete living cells or their components (e.g., bacteria,
enzymes, chloroplasts) to obtain desired products is referred to as bioprocessing.
• Bioprocessing is also essential to the production of renewable biofuels like ethanol
and biodiesel, therapeutic stem cells, gene therapy vectors, and new vaccines, among
other emerging industries and technologies.
• Bioprocessing includes two important processes - Upstream and downstream
processes.
Cell Treatment Bioprocessing
Cell treatment bioprocessing is an order that combines the fields of cell treatment and
bioprocessing. Cell treatment bioprocessing aims to build up reproducible and sturdy
manufacturing procedures to produce remedial cells. Commercially significant bioprocesses
can:
• Generate items to keep up the entirety of the quality gauges of biopharmaceutical
drugs
• It can also offer clinical and industrial measures of remedial cells all through different
phases of improvement.
• Control the expense of merchandise (CoGs) of the last medication item.
Upstream Bioprocessing
• The process of converting raw materials into a form that can be used in a biologic
manufacturing process is known as upstream bioprocessing.
 • Harvesting and purifying natural products, culturing cells, and producing recombinant
proteins are all examples of this.
• Upstream bioprocessing's goal is to create a high-quality starting material for
downstream bioprocessing.
The upstream process is one of the two main parts of a bioprocess. It includes the initial steps
of a fermentation process. The preparation of microorganisms is the first step of upstream
bioprocessing. The desired microbes should be isolated and selected for the bioprocess. Then
they should be cultured in a suitable growth medium. Medium preparation is the second step.
Optimal conditions, together with the nutrients, are supplied for the growth and multiplication
of microorganisms inside a bioreactor.
Downstream Bioprocessing
• The term "downstream bioprocessing" refers to the steps that take place after the
initial bioprocessing steps, which involve the production of a biological agent.
• There are three main steps involved in downstream processing- Purification,
Formulation, and Storage of the agent.
Downstream bioprocessing refers to the several final steps of a bioprocess that
involve product harvest. The downstream process starts when the product
development is completed. The steps include extraction, purification, and packaging
of the final bioproduct of the bioprocess. Downstream bioprocessing is also known
as product recovery. Amino acids, antibiotics, organic acids, vitamins, and vaccines
are some of the bioproducts resulting from a bioprocess. The desired quality of the
bioproduct is achieved during the downstream process. Volatile products can be
purified using a distillation process. Moreover, biomass separation can be done by
centrifugation. Therefore, the stages involving separation, extraction, purification, and
polishing belong to downstream bio processing.
The downstream process involves the following steps-
a. Elimination of Insolubles - By soaking a sample of the material in a solvent, insoluble
impurities are removed. Insoluble impurities will sink to the bottom of the solvent, while the
rest of the material will dissolve. After that, the solvent can be poured off, leaving behind the
insoluble impurities.
b. Isolation of Product - Product isolation is a chemical engineering technique for isolating a
product from a mixture. The product is extracted using a solvent after the mixture is placed in
a container. The product is then isolated after the solvent has been removed.
c. Product Purification - The removal of impurities from a product is known as purification.
Distillation, chromatography, and crystallization are some of the methods that can be used to
accomplish this. Purification aims to create a product that is free of contaminants and pure.
d. Polishing of Product - The product will be polished after it has been cut to remove any
scratches or blemishes that occurred during the cutting process.

Scale up of Fermentation Processes

Definition: - Scale up means the art for designing of large scale apparatus or full size plant
(prototype) using the data obtained from the laboratory studies

Objectives: - To provide master manufacturing formula

• To identify the critical features of the process

• Evaluation, Validation and Finalization of process

• Guidelines for production and process control

• Review of the processing equipment

• To produce physically and chemically stable products.

Requirements for scale up process

• Personnel Requirements

• Space Requirements

• Administration and Information Process

• Physical Testing Area

• Standard Pilot-plant Equipment Floor Space

• Storage Area

• Review of Formula

• Raw materials

• Equipment
• Production rates

• Process Evaluation

• Preparation of Master Manufacturing Procedures

• Product Stability and Uniformity