Download department of biological sciences

Commercialization of Biotechnology
Products
BIOT-305
Topic: Production of Citric Acid at Commercial
Level
GROUP LEADER
Hafiza Madeha Khaliq
(11-10033)
1
Introduction:
Citric acid is one of the most important organic compounds which are produced by
fermentation. It is widely used to impart a pleasant, taste and flavor to foods and
beverages. It also has a function as an additive in detergents, pharmaceuticals, cosmetics
and toiletries. On average, 64% of citric acid produced is used for food and beverages,
22% for detergents and cleaning products, 10% for pharmaceuticals and nutritional
products, 2% in cosmetics and toiletries while rest of the 2% is used for other different
purposes. Citric acid is the component of bones and it is used in food coloring to balance
the pH value of a normally basic dye because of its acidic nature and buffering capacity.
History:
Wehmer observed the citric acid fermentation for the first time in 1893 as a fungal
product by a culture of Penicillium glaucum on sugar medium. Then he isolated two new
fungal strains having the ability to accumulate citric acid after few years but the
contamination problem and long duration of fermentation made the industrial production
of citric acid unsuccessful. In 1916, Currie found various strains of Aspergillus niger that
had ability to produce significant amount of citric acid. Two other most important
findings were:
1) Aspergillus niger grows well at pH value around 2.5 to 3.5
2) High concentration of sugar favors citric acid production.
As a result, first citric acid fermentations were carried out in surface culture in 1930.
Some units were implanted in England, Soviet Union and Germany for commercial
production.
World Production and Consumption of Citric Acid in
2009:
Pie chart given below shows the world consumption of citric acid.
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Citric acid is produced and consumed through out the world. The data shown above is of
year 2009. Worldwide production of citric acid is around 1,600,000 tones per anum. The
majority of the production and consumption in 2009 was in Western Europe, US and
China. It is estimated that nearly 80% of the global citric acid is consumed by Western
Europe, Eastern Europe, US, China, Central Europe and Asia. The citric acid industry is
continuously influencing by an increase supply form China in an abundant global
capacity.
In Asia, the consumption is highest in India, Indonesia, Thailand and republic of Korea.
More than 50% of the citric acid is produced in China and production of citric acid in
china is mainly exported to United States, Japan, Belgium, Netherlands, India, Spain and
other countries.
Natural Sources of Citric Acid:
Citric acid is found in variety of citrus fruits, vegetables. Lemon and lime have higher
concentration of citric acid which is about 47g/litre in juices of these fruits. Krebs cycle
is the natural process of production of citric acid in all living cells.
Krebs Cycle:
Krebs cycle is a series of enzyme catalyzed reactions that occurs in cytosol of
prokaryotes and mitochondrial matrix of eukaryotes. This metabolic pathway is actually
one of the three major pathways of cellular respiration. The other two are glycolysis and
electron transport chain. It is also called citric acid cycle or tricarboxylic acid cycle
(TCA).
Pyruvate is produced at the end of glycolysis which is an energy rich compound and
plays important role in ATP synthesis. It enters in Krebs cycle through a transition
reaction that connects glycolysis with Krebs cycle. In this reaction pyruvate is converted
into acetyl-coenzyme A and carbon dioxide is released. This is an oxidation reaction.
Electrons are also released form pyruvate. This acetyl-CoA which is a two carbon
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compound joins with a four carbon compound called oxaloacetate. As a result citric acid
is produced in the first step (a six carbon compound) that’s why Krebs cycle is called
citric acid cycle. This cirate is then converted into isocitrate which is then oxidized and
decarboxylated to a five carbon compound called alpha ketoglutarate. During this
reaction, most of the electrons released are accepted by NAD and NADH is formed
which carries electron to the electron transport chain and carbon dioxide is released.
Alpha ketoglutarate is oxidized and decarboxylated to succinyl-CoA. This step produces
NADH and Carbon dioxide. Succinyl-CoA is a high energy compound which has a high
energy bond coupling the Coenzyme A component to the succinate. When this bond
breaks, it releases its high energy to make a high energy phosphate bond which is used to
form ATP by substrate level phosphorylation. Infect, succinyl-CoA reacts with GDP
(guanosine diphosphate) to form GTP (guanosine triphosphate) which in return reacts
with ADP to make ATP. At the end, succinate is formed which is a four carbon
compound. Succinate is then dehydrogenated and electrons are released which are
accepted by FAD to produce FADH which also provide electrons to the electron transport
system. Fumarate is the product of this reaction which is a four carbon compound.
Fumarate is converted into malate (a four carbon compound) in the presence of water.
Malate is oxidized and oxaloacetate is formed and this oxaloacetate serves as a substrate
for the restart of the Krebs cycle.
In Krebs cycle, at each step a different enzyme is used as shown here:
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The input and output of Krebs cycle per glucose molecule as follows
Input
2-acetyl groups
6 NAD+
2FAD
2ADP+ 2Pi
Output
4CO2
6NADH
2FADH2
2ATP
 How Could be Krebs Cycle used for Citric Acid
Production?
As citric acid is one of the intermediates that is produced in TCA cycle so if use the
whole cycle for citric acid production, it increases cost and leads to loss of resources and
time. In order to maximize the citric acid production, we can follow various points;
o Mutation cause by using various mutagens for example gamma radiations in the
organism used can be helpful.
o Inhibiting the free flow cycle by changing the environmental conditions e.g.,
temperature, PH and media composition can be another effective way.
o Removal of the metal ions that act as co-factors for various enzymes used in
unnecessary steps of Krebs cycle cab also be fruitful.
o Using inhibitors for enzymes used in unnecessary steps of the Krebs cycle is
another good option.
Microbial Production of Citric Acid:
There are number of microbes that have been used for citric acid production. Some of the
examples are;
i.
ii.
iii.
iv.
v.
vi.
vii.
viii.
ix.
x.
Arthrobactor paraffines
Bacillus lichenopromis
Corynebacterium ssp.
Aspergillus niger
A. acucleatus
A. Carbonarius
A. awamori
A. foetidus
A. fonsecaeus
A. Phoenicis
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xi.
xii.
xiii.
xiv.
xv.
xvi.
xvii.
Penicillium janthinellum
Candida tropicalis
C. oleophila
C. guilliermondii
C. citroformans
Hansenula anamola
Yarrowia lipolytica
However most of these are unable to produce citric acid commercially.
Microorganisms of choice: “Aspergillus niger”:
Aspergillus Niger is a fungus that is included in Aspergillus subgenus, Circumdati. It is
considered as the organism of choice for the commercial production of citric acid because
of a number of reasons.
i.
ii.
iii.
It produces more citric acid per time unit so it provides high yield of citric acid.
It is easy to handle.
It has ability to ferment a verity of cheap raw materials.
Aspergillus Niger is a fungus that is one of the most common species of genus
Aspergillus. In 2006, the World Health Organization found that Aspergillus Niger derived
organic acids are not only safe to consume but they also have anticarcinoganic and
antiangiogenic (tumor reducing) properties for instance actibind.
 How to grow Aspergillus niger in Laboratory?
The US department of Agriculture’s Agriculture Research Service Culture Collection
maintains several strains of Aspergillus niger. After getting that following steps are
followed;
i.
ii.
iii.
iv.
v.
vi.
vii.
viii.
39 gm of agar is dissolved in 1 liter water in a clear beaker.
Mixture is heated to dissolve the agar completely.
It is stirred with a sterile glass rod.
Mixture is sterilized for 15 minutes at 15psi (pounds per square inch).
Mixture is cooled approximately to 45 Celsius.
It poured it into sterile Petri dishes and allowed to set.
It is inoculated with Aspergillus niger by wiping a swab containing the
culture across the surface of agar.
Inoculated petri dishes are stored upside down at 25 Celsius with the agar
on the upper surface. Substantial colonies take 5 to 7 days to develop.
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 Side effects of using Aspergillus niger:
Aspergillus Niger is a known allergen that causes allergic reactions if inhaled. According
to the environmental protection agency at epa.gov, the antigen that causes lung
hypersensitive and produces these reactions is found in the strains which are used in
industrial applications. In severe cases, A. niger also causes asthma and aveolitis
(inflammation of the lung’s air sac or alveoli). The fungus Aspergillus niger can also take
root inside the human body for example pulmonary cavity where the wind pipe branches
off into lungs. This condition is termed as aspergillosis and can have terrible side effects
for example chronic coughing and coughing of blood (hemoptysis) and it commonly
occurs in individuals suffering from immune system deficiency. This problem can be
solved by using mutant strains of A.niger.
 Improvement of Strains:
The improvement of strains for enhanced citric acid production has been carried by
mutagenesis. Different mutagens are used to induce mutation in parental strains and
mutants of Aspergillus niger are used for commercial production. Hyper producer strains
can be obtained by using ultra violet radiations and chemical mutagens together.
Different methods of fermentation used can produce different yields of citric acid even if
same strain is used. For example, in Institute of Food and Production Biology in Dhaka,
several high citric acid producing strains of Aspergillus niger have been prepared with
the help of gamma radiations. These mutant strains have been found to yield three to five
times more citric acid from molasses then their parent strain (CA16).
Substrates:
Citric acid is produced by fermentation using inexpensive raw materials using crude
natural products such as hydrolysate starch, sugarcane broth and byproducts like
sugarcane and beet molasses. Molasses is preferably used as a source of sugar for
microbial production of citric acid due to its relatively low cost and high sugar content
(40 to 55%). Since it is a byproduct of processing of sugarcane, grapes or sugar beets into
sugar, that’s why its quality varies considerably. Main carbohydrates are glucose,
fructose, raffinose and some other oligo- or polysaccharides. All types of molasses are
not suitable for citric acid production. The composition depends on various factors;
i.
ii.
iii.
iv.
Variety of beet and cane
Method of cultivation
Condition of storage
Transport
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Both beet and cane molasses are suitable for citric acid production, however, beet
molasses is preferred due to its lower content of trace metals and it supplies better
production yield than cane molasses. In case of cane molasses, it contains some metals
such as iron, calcium, magnesium, manganese, zinc etc. which retard citric acid synthesis
and that’s why it requires some pretreatment for the reduction of these metals, which
increase the cost. So beet molasses is preferred and addition of phytate at the beginning
of incubation of beet molasses results in about three fold increase in citric acid
accumulation. Phytate is a stored form of phosphorus in plant tissues especially in brain
and seeds.
 Citric acid production techniques:
Citric acid production by fermentation is most economically and widely used way of
obtaining this product. More then 90% of the citric acid produced in the world is obtained
by fermentation. It has certain advantages:
i.
ii.
iii.
iv.
Operations are simple and stable
Plant is less complicated as it requires less sophisticated control system
Lower energy consumption is needed
Power failures don’t critically effect the functioning of the plant
In this process, organism is grown in liquid medium which is vigorously aerated and
agitated in large tanks called fermenters. This liquid media is a rich nutrient medium
required by A.nigar to grow. The growth conditions maintained are different according
to the organism used i-e. Mutant A.nigar require different conditions to grow as compare
to the natural A.nigar. So in batch fermentation, A.nigar is grown in a know amount of
culture medium for a defined period of time and then cell mass is separated from the
liquid before further processing .While in continuous culture, the mass is separated
depending on rate of product formation and inflow of fresh medium. As batch culture is
used for citric acid production, so it requires 5-15 days depending on the process
conditions.
 Phases:
Citric acid production by fermentation can be divided into three phases:
i.
ii.
iii.
Preparation and incubation of raw material
Fermentation
Recovery of the product
 Fermentation types:
The industrial citric acid production can be carried out in three different ways;
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i.
ii.
iii.
Submerged Fermentation
Surface Fermentation
Solid state Fermentation
 Methods of choice: “Submerged fermentation”
This technique is widely used for citric avid production and about 80% of world
production is obtained by submerged fermentation. Batch or continuous, both can be used
for citric acid production but batch culture is more frequently used.
 Chemical requirements:
i.
Carbon Source :
Presence of carbohydrates is essential for good production of citric acid. Type and
concentration of carbon source is really important because it must be easily
metabolized. Sucrose is more suitable carbohydrate. Glucose, glactose and fructose
can also be used. Cost and need of pretreatment must be taken into account for
choosing carbon source. Molasses has some trace elements which must be pretreated
by Potassium Ferro cyanide.
ii.
Phosphorous Source:
The level of phosphate has positive effects on citric acid production. Phosphorous
level affects the level of enzyme activity. But excess of phosphorous leads to a
decrease in CO2 fixation which in turn increases the sugar acids formation instead of
citric acid. Potassium dehydrogen phosphate is used as phosphorous source.
iii.
Nitrogen Source:
Nitrogen source is also required for citric acid production. Concentration and nature
of the nitrogen source needs to be taken care of.
Preferred nitrogen sources include:
i.
ii.
iii.
iv.
v.
Urea
Ammonium nitrate
Sulphate
Malt extract
Peptone etc.
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Acid ammonium compounds are preferred as they decrease the pH which is important
fir citric acid production. Concentration of nitrogen source requires ranges from 0.10.4g/L. But high nitrogen concentration increases fungal growth and sugar
consumption. As a result, citric acid production is decreased.
iv.
Trace elements:
Presence of trace elements has a significant impact on citric acid accumulation by
A.niger. Divalent metal ions such as zinc, manganese, iron, copper, magnesium etc
increase the citric acid production when used in trace amount. Control of these trace
elements is very important to get maximum yield.
v.
Lower Alcohols:
Lower alcohols e.g. methanol, ethanol, n-propanol, iso-propanol, methyl lactate etc. when
added to the crude carbohydrates enhance the production of citric acid by neutralizing the
negative effects of metals. Alcohols principally act on the membrane permeability in
microorganisms and affect the phospholipids composition by disrupting the ion channels
in plasma membrane. As a result balanced osmotic environment is disturbed.
vi.
Antifoams:
These are the substances which are used to reduce the foam formation during citric acid
production. Oils and fats e.g. linseed oil, tung oil, linolenic acid etc. are used as antifoams
which control the foam formation, maintain the fermentation process and enhance the
citric acid productivity.
 Physical Factors:
i.
pH:
Maintaining the optimum pH is very important to get maximum production of citric acid.
pH of a culture changes in response to the microbial activities. Secretion of organic acids
such as citric acid is one of the most obvious reasons which cause the decrease in pH. So
change in pH kinetics is mainly dependent upon microbes. Nature of the substrate and
production technique also influences the pH. Buffer solutions are used to maintain the
optimum pH (3.5-4.5).
ii.
Aeration:
Citric acid production is an aerobic process, so proper oxygen supply is required for this
purpose. Increased aeration rate enhances the yield and reduces the time period required
for the completion of the process. It is important to maintain the dissolved oxygen
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concentration as its reduction or absence can be highly harmful. Proper aeration devices
are required to fulfill this demand. Oxygen demand is dependent upon the viscosity of
fermentation broth. However, high aeration rate can lead to the high foam formation
during the extensive microbial activity. Antifoaming agents are used to avoid this
problem e.g. oils and fatty acids.
iii.
Temperature:
It is important to maintain the optimum temperature during the fermentation process for
the production of citric acid that ranges from 30-32 Celsius, depending upon the type of
organism used.
 Fermenter Design:
Stainless steel fermenter is used for citric acid production as it does not corrode or affect
the microbes and fermentation products. It can easy be cleaned. All the nutrients and
microbes are put into the fermenter and air is bubbled through to assist the microbial
aerobic respiration. Gas outlet is there which helps to release the pressure build up inside
because of CO2 production. A water or cooling jacket is there to maintain the optimum
temperature as temperature rises because of different microbial activities. Paddle stirrers
are there to provide proper agitation, and to make sure that nutrients are distributed
evenly. Temperature, pH and oxygen are linked to a computer which monitors all the
parameters or conditions inside and makes it easy to maintain them. There is an outlet at
the end from where the product is run off, and it is separated from the microbes and
purified for sale and distribution.
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 Extraction of citric acid
After the fermentation process the next step is the extraction of citric acid from
fermenters. The extraction begins with the filtration of broth until it is cleared. Calcium
citrate is precipitated by the addition of magnesium-free. Since magnesium is more
soluble then calcium, some acids may be lost in solution as magnesium citrate if
magnesium is added. Then in the next step calcium citrate is filtered and the filter cake is
treated with sulfuric acid to precipitate the calcium. The dilute solution containing citric
acid is purified by treatment with activated carbon and passing through iron exchange
beds. The purified dilute acid is evaporated to yield crystals of citric acid.
Further purification may be required according to our own requirement. From here the
citric acid is packed and transported to the different locations
 Applications of Citric acid:
The citric acid is a naturally occurring weak acid. Its weakness makes it extremely
beneficial and useful for all manner of applications as it can be easily handled and
ingested by both people and animals.
 Citric acid is mainly used as flavor enhancer for beverages.
 It helps in tenderization of the meat by breaking its collagens down to more
palatable form.
 It is an important ingredient of confectionaries and other food products.
 It acts as an antioxidant also for the readymade food e.g. tinned meat, vegetables
and fruits to preserve them.
 It is used in the manufacturing of detergents for the cleaning purpose. It attracts
and removes the dirt and many other stuck-on messes from the cloth.
 Citric acid is a good buffer so is used in household cleaners to control PH.
 It can chelate the metal ions so it is also useful in soaps and laundry detergents by
letting them to produce foam and work efficiently.
 I t is also beneficial for the purpose of water softening.
 It is also used for the purpose of drafting and designing e.g. ferric ammonium
citrate is used to produce blueprints. This paper is placed under any drawing,
when paper reacts with light, the ferric ammonium citrate changes to ferrous salt
as a result blue color is produced.
 In photography, the sodium citrate, with the help of other chemicals transforms
the light-responsive areas into photographs. After the photo paper that is coated
with light sensitive chemicals is exposed to light through a negative is placed in a
series of tubs of citric acid that forms the picture on photo paper from negative.
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




It is also an important constituent of pharmaceuticals and especially diet
supplements e.g. vitamin supplements. Citric acid or its salts enable the minerals
in the supplements to be biologically available for the body to use up.
It is very important in the treatment of alcoholics so it can also be used to
minimize the negative effects of alcohol e.g. in perfumes, cough syrups, shaving
lotions etc.
Citric acid prevents blood clotting so, it is very helpful in open-heart surgery to
stop heart contractions and make the surgery easier to perform. Potassium citrate
is used for this purpose.
It also acts as a binding agent in biodegradable detergents.
It is used to mask fowl smelling also.
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Company Profile
 Management Team:
The management of ‘Premium Citrus Company includes;
Dr. Kausar A.Malik
Mr. Syed Ahmad Hashmi
Hafiza Madeha Khaliq
Samraiz Hafeez
Faiza Asghar
Haider Khaliq
Moaaz Aqeel
Alina Rafi
Madiha Mehmood
Sohail
Murtaza
Raafia Ramzan
Sehrish
Process Consultant
Marketing Consultant
Chief Executive Officer (CEO) & Project manager
GM of Sales & Marketing
GM of Production
Recovery Manager
Quality Controller
Auditor
HR Officer
Labour officer
Finance Manager
Sales Representative
Sales Representative
 Compensation and Ownership:
Premium Citrus Company functions as an enterprise. The salaries of the management
team are listed below.














Profit goes to owners.
Process Consultant
Marketing Consultant
Chief Executive Officer (CEO) & Project manager
GM of Sales & Marketing
GM of Production
Recovery Manager
Quality Controller
Auditor
HR Officer
Labour officer
Finance Manager
Sales Representative
Labour
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Rs. 200000
Rs. 200000
Rs. 250000
Rs. 150000
Rs. 150000
Rs. 150000
Rs. 100000
Rs. 75000
Rs. 50000
Rs. 30000
Rs. 75000
Rs. 20000
Rs. 7000-15000
 Board of Directors:
Dr. Kausar Abdullah Malik
Mr. Syed Ahmad Hashmi
Dr. Hamid Saeed
Prof. Athar Azeem
Process Consultant
Marketing Consultant
Environmentalist
Sociologist
 Contracts and Agreements:
Premium Citrus Company Private Ltd. has the following certificates and agreements on
file;











Kohinoor Sugar Mills Limited, Company Lahore.
R&L International (Shanghai)Co.Ltd
CHANDRASHEKHAR Exports PVT. Ltd.
Unique Biotech Ltd. India
Sabro technologies private limited Pakistan.
Pakistan Beverage Ltd.
Popular Food Industries Ltd.
Shan Food Industries. (Pvt.) Ltd.
Abbott Laboratories Pak Ltd.
Pakistan Refinery Ltd.
Analytical Technologies Limited.
(Molasses)
(Fermenters)
(Corn steep liquor)
(Aspergillus niger)
(Electric equipment)
Infrastructure
 Banking:
Pakistan Industrial Credit and Investment Corporation (PICIC) Commercial Bank
Limited. Fees are based on the type of transaction.
 Insurance:
State Life Insurance Corporation of Pakistan.
Premium citrus company had product liability insurance policy of Rs. 3,000,000 through
Pakistan General Insurance Company Limited with a premium of Rs. 30,000 per month.
 Capitalization:
15
Premium Citrus Enterprise relies on National Business Incubation Association (NBIA) to
fund its operations and capital expenditures. The company's future capital requirements
will depend on numerous factors including its own efforts and the efforts of its
collaborative partners to commercialize its product.
Product and Service plan
 Purpose of the Product:
Citric acid of Premium Citrus has an important use in, hospitals, home care units, food,
beverages, chemical industries and other health care facilities. Our basic purpose is to
make Pakistan self sufficient in citric acid production for its progressive economy.
 Unique Features:
Citric acid, product of Premium Citrus Company, is useful in shampoos, soaps and
laundry detergents as a cleaning agent. It is also used in pharmaceutical, food and
beverage industries as a buffering and flavoring agent respectively. It enhances the shelf
life of the products so it is a good preservative. It is an important ingredient of dairy
products e.g. cheese etc. The citric acid in lemon juice can cure heartburn, bloating and
belching, aids digestion and prevents constipation.
 Stage of Development:
Premium Citrus Company Pvt. Ltd. has been founded in 2010 and its corporate offices
are in Pakistan. Its first commercial citric acid has been produced in May 2011. Premium
Citrus Company is a chemical company that manufactures and markets all pure citric acid
by patented fermentation technology. Our customer market includes cosmetic, food,
medicine, detergent and beverage market at national as well as at international level.
 Future Research and Development:
Premium Citrus Pvt. Ltd. believes that its products’ functionality will make them
potential candidates for future development in its manufacturing technology. We have an
access to leading authorities in microbiology, industrial biotechnology, and fermentation
technology and govt. agencies so in future we intend to provide facilities for the
experimentation, analysis and technological innovations at the basic research level.
 Trademark, Patent and License:
Premium Citrus Company uses trade mark, patent and trade secrets to exclude others to
know its proprietary technology. We had filed applications in Initial Public Offering and
got utility patent in Pakistan regarding the composition of medium, and the fermentation
technology that has been used to manufacture citric acid. Premium Citrus Company also
believes that its trademark, trade name is valuable asset and is under protection.
 Product and Service Limitations:
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Contact with dry citric acid or with concentrated solutions can result in skin and eye
irritation, so protective clothing should be worn when handling these materials. Products
of citric acid may not be for extensive or regular use. They can give some people
heartburn, nausea, vomiting, and stomach pain and can also damage the enamel on their
teeth. In some cases, there may be confusion, anxiety, weakness, irritability and mood
changes. Customers should consult a physician before consuming such products
extensively.
 Production Steps:
Production of citric acid by Premium Citrus Company follows the steps given below;
 Procurement of raw materials
 Growing A. niger in Laboratory
 Citric acid Production by batch submerged fermentation
 Harvesting of Product
 Filtration
 Disposal of wastes
 Purification of product
 Recovery
 Drying or crystallization of citric acid
 Final formulation
 Testing (3-5 days) and Quality control (7-10 days)
Note: The Quality Control Department has the authority to release or discard the
product.
 Packaging and Labeling
 Shipping after release from quality control or Storage.
 Advertisement and Marketing
 Facilities:
Premium Citrus Company believes that all of its manufacturing plants, laboratory
facilities, and laboratory equipment are in satisfactory condition and are appropriate for
the purposes for which they are under use.
 Suppliers:
Premium Citrus Company has annual purchase contracts for raw materials and
manufacturing equipments. Fermenters are imported from U.K Company R&L
International (Shanghai) Co. Ltd. Aspergillus niger kit is obtained from Indian company
named as Unique Biotech Ltd. Premium Citrus Company has got raw molasses from
Kohinoor Sugar Mills Limited, Company Lahore. Plastic ware and glassware are obtained from PIN TECH INSTRUEMENTS Pakistan and Analytical Technologies
17
Limited. All these supplies are based not only on the lowest price but also on timely
shipping.
 Environmental Factors:
Citric acid is recognized as safe for use in food by all major national and international
regulatory agencies as it is naturally present in almost all forms of life and its excess
amount is readily metabolized and eliminated from the body. All waste materials
produced at the end have check and balance system. Waste materials released in the
environment have values lower than those of NEQ values so our production plant is
environmental friendly.
Market Plan
 Current size:
Premium Citrus Company covers an area of 3 acres, currently.
 Growth potential:
Premium Citrus Company is going to be the first production company for citric acid in
Pakistan. All local and industries including food, detergent, beverages, and wherever the
citric acid is used depend upon foreign countries for its supply. We believe that our
revenue opportunity would be huge especially at national level. Premium Citrus
Company also aims to export its high quality citric acid in the foreign market. The
company also hopes to have marketing arrangements with other well-placed companies
by the end of 2011.
 Geographical Locations:
Premium Citrus Company is located near located in industrial area present near the EME
housing society, Lahore and product is supplied primarily to the domestic and foreign
distributors as well as industries.
 Trends:
As consumption of citric acid in Foods, personal care products and especially in
beverages is increasing day by day, so opportunities are arising for citric acid
manufacturers too to increase its supply for more profit.
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 Seasonality factors:
Citric acid is mostly used in beverage industries and beverage industry production rate
increases in summer season so citric acid demand also increases in summer. Rise and fall
graph of demand and supply of citric acid also reveals the same thing.
 Profit:
The sale of citric acid directly to the beverage industries carries the highest profit margin.
Citric acid sold to the domestic beverage, food and pharmaceutical companies carries an
average profit of 20% while 35- 40% average profit has been estimated on the foreign
export.
 Distribution:
Premium Citrus Company has made an exclusive distribution agreement covering all
companies in Pakistan.
 Competitors:
As Premium Citrus Company is the leading provider of citric acid in Pakistan, so recently
it’s not facing any competitor from inside but competition is intense at the International
level. The major competitors of Premium Citrus Company include Asian countries, India,
China, U.S., U.K. and Western Europe in terms of product quality and sale in Pakistan.

Customers:
Every person and every industry that is willing to use our product are our customers.
 Target Market:
Target Market of Premium Citrus Company includes beverage, pharmaceutical, food
industries, cosmetic and chemical companies and detergent manufacturing industries.
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 Pricing Profile:
Premium Citrus Company also provides discount for annual contracts. Introductory offers
are available during the first year of the product being marketed.
Price of the product=Rs. 200 /kg
Market Penetration
 Sales Representatives & Direct Sales Force:
Premium Citrus Company has a marketing department with a general manager and
twenty sales representatives who promote the product sale using a number of direct and
indirect ways.
 Direct mail/Telemarketing:
Premium Citrus Company has employed telemarketers to reach that market where reach
of direct sales force is not possible.
 Advertisement:
Premium Citrus Company is using the internet facility to market its product. The website
is www.pakcitricacid.com.
 Packaging and Labeling:
Premium Citrus Company has a trademark of muscular orange with name of the company
“Premium Citrus” on its label written in its unique style. Labeling depends on type of the
material being used for the packing. Citric acid is packed in plastic packets and
containers. Packaging stuff is environment friendly. Each packet or container has
complete information of proper use of the product, its limitations, company contact
information, and supporting detail.
 Services and Warranties:
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Premium Citrus Company will fully refund when the unused product is returned with a
written description. Toll free number is also available for both technical information and
customer service assistance.
 Receiving Orders:
In our company orders are taken by Phone, fax, e-mail and mail-in orders are received in
the Customer Service Department. Then these orders are entered into computer for
immediate processing. Once the product has been produced it is checked by quality
control department to maintain the best quality and to avoid complains.
 Billing the Customers:
Customers are billed net 30 days with a 1% discount for orders paid within 10 days. A
credit check is done on all new customer accounts.
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Paying the Suppliers:
We pay the suppliers 20% before the supply and 80% after one month until we don’t
check the quality of raw material. Because our whole process of fermentation depend
upon the quality of raw material provided. If the raw material is not according to our
requirement our whole process will become useless.
 Reporting to Management:
Weekly staff meetings are held by the General Manager of each section of the company,
such as research & development, administration, and sales and marketing. Employees are
to report all incidents of concern to their respective GM. It is then the responsibility of
that GM to inform the CEO to determine if any action is to be taken.
 Handling Warranties and Returns:
The company maintains both Technical Service and Customer Service Departments.
Performance-based customer concerns are handled by the Technical Service Department.
The Customer Service Department handles non-performance customer concerns, such as
product leakage, mishandled orders, frozen or overheated product, short orders, etc.
 Security Systems:
Sensitive documents are stored on site in a fireproof locked cabinet. Back-up tapes of
computer Information are made nightly and duplicates are stored off site. Petty cash is
stored in the company safe. An alarm system with password entry has been installed to
avoid any type of inconvenience.
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 Product Development:
New product development occurs when requested by customers. We can use different
strains of A.nigar to have better yield and good quality, we can also cause mutations in
A.nigar e.g. by exposing it to radiations, we can also insert new gene in this organism by
genetic engineering which may help in increasing the yield.

Manufacturing:
The formation of product takes 5- 15 days. Once the product is made, it is taken to the
Quality Control Department for sterility and performance testing.
 Planning Chart: “Market penetration”:
The GM of sales & marketing in collaboration with the CEO set the strategy for market
penetration. This strategy will include, but will not be limited to, the decision to use a
network of distribution, establishment of sales territories, sales-force training and a
schedule detailing the timing of events.
 Risk analysis:
There can be no assurance that our product will continue to be successfully
commercialized and accepted for use by people. Because majority of Pakistanis are crazy
about products imported from foreign countries. Moreover, there can be no assurance that
the interests and motivations of any corporate partner, distributor or licensee would be or
remain consistent with those of New Age BIOTECH, or that such partner, distributors or
licensees would successfully perform the necessary technology transfer, clinical
development, regulatory compliance, manufacturing, marketing or other obligations.
 Support to the economy:
The most important purpose of establishing a company in Pakistan is to provide people
with reasonable jobs. It will definitely reduce unemployment and raise our economy.
Pakistan will also become independent in production of quality citric acid.
 Cost Analysis:
Profit goes to owners
Pays:
Process Consultant
Marketing Consultant
Whole profit
Rs. 200000
Rs. 200000
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Chief Executive Officer (CEO) &
Project manager
GM of Sales & Marketing
GM of Production
Recovery Manager
Quality Controller
Auditor
HR Officer
Labour officer
Finance Manager
Sales Representative
Sales Representative
Labour
Materials:
Molasses
Potassium dehydrogen phosphate
Corn steep liquor
Stainless steel fomenters
Land
Building construction
Rs. 250000
Rs. 150000
Rs. 150000
Rs. 150000
Rs. 100000
Rs. 75000
Rs. 50000
Rs. 30000
Rs. 75000
Rs. 20000
Rs. 20000
Rs. 7000-15000
Rs. 340000/ ton
Rs. 85975/ ton
Rs. 9350/ ton
Rs. 2,550,000/ Per
fermenter (25 fermentors)
Rs. 240,000,000
Rs. 50,000,000
Total Fixed Cost=Rs. 353750000
Variable cost=Rs. 355667325
Dead stock value of production=10 tons/day
Price of the Product= Rs. 200 /kg
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Biosafety Regulations
There are certain laws for the safety of environment. It’s important for industries to obey
them. These laws are for the prevention of environmental hazards. In order to save the
environment and keeping the environment healthy, laws were made as the
Environmental Protection Act 1997.
As the country progresses, the industries become more populated with the
production. And as a result of this production, there is a mass increase in the pollution
and hazards to the environment also increase.
We the owners of Premium citrus Pvt. Ltd obey these laws.
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Our industry is located on the Multan road-18 km far from Multan road which is
an industrial area. The habitation in the surrounding of the factory is very low and
no one is directly affected or disturbed by the company.
The machinery like fermenters is noise proof so there is zero noise pollution
because of the factory.
The working of the machinery is totally environmental friendly.
The waste produced as a result is majorly in the form of liquid which is released
in the environment after passing through a proper treatment channel of the
factory.
Composters are present in the factory area where the solid waste is treated.
The gases and smoke that are released in the environment have the toxicity value
less than the NEQ values. Similar is the case with noise and liquid waste
produced.
The microorganism that is used for the fermentation process is “Aspergillus
niger” which is a mutant organism that produces 3-5 times more citric acid than
the ordinary A.niger.
There are biosafety laws regarding genetically modified organisms; they can
undergo changes in their genetic makeup and as a result they can produce
secondary metabolites harmful for the environment. The organism we are using is
GRAS (generally regarded as safe) which means it does not have any hazardous
environmental effects.
Our staff and management are completely aware of the rules required for working
in a laboratory and obeying them as well by fulfilling all the biosafety.
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