single cell protein2

SINGLE CELL PROTEINSITS PRODUCTION

From Chlorella and Spirulina

A Powerpoint presentationBy: Mariadoss

SINGLE CELL PROTEIN• The dried cells of microorganisms(algae,bacteria,fungi)used as food

or feed are collectively known as “microbial protein”.

• The term ‘microbial protein’ was replaced by a new term ‘single cell protein’ during the first international conference on microbial protein held in 1967 at the Massachusetts Institute of Technology(MIT),Cambridge.

• The term single cell protein (scp)refers to mixed protein extracted from pure or mixed culture of algae, yeasts, fungi, or bacteria used as a substitute for protein rich food in humans and animal feeds.

• The term SCP was coined in 1966 by Carol L. Wilson at MIT

SPIRULINA

• Is a microscopic blue green algae in the shape of a spiral coil.

• Found in fresh waters, sea, marine waters, inland saline lakes

• Common name for the human and animal food

• Produced by two species of cyanobacteria:• Arthrospira platenis and A.maxima

Spirulina maxima

s.fusiformis

CONSTITUENTS OF SPIRULINA SCP

A.MAJOR constituent• Crude proteins 65%• Carbohydrate 16%• Lipids 6.7%• Nucleic acids 4.2%

B.VITAMINS• Biotin 0.2%• Cyanocobalamin 66.0mg• Folic acid 18.0mg• Riboflavin 1.8mg• Thiamin 0.12mg• Tocopheol 0.81IU• β-carotene 320,000IU

MINERALS• Calcium 6.6mg• Phosphorus 0.9gm• Iron 4mg• Sodium 0.8mg• Potassium 1.3mg

ESSENTIAL AMINO ACIDS• Lysine 2.99%• Cystine 0.47%• Methionine 1.38%• Phenylalanine 2.87%• Threonine 3.04%

MASS CULTURE OF SPIRULINA

Spirulina is cultured in large scales in artificial ponds or tanks and oxidation ponds.

• Open Culture1. open circulating system2. oxidation pond system• Closed Culture1. Photobioreactor2. Greenhouse

1.OPEN CIRCULATING SYSTEM

• An open circulating system is a man made open tank or shallow pond.

• It may be circular or rectangular in shape.• The depth of the tank should be in between

25 and 30 cms.• Size of the tank may be 500m2-500m2.• It is built with brick or concrete and the interior is

lined with a sheet of polyvinyl chloride(PVC),

Circular tank

• In a circular tanks a stirrer with a rotating arm is kept at the centre to provide enough stirring for the culture.

• Usually the culture tanks are kept open while functioning.

• Sometimes, they may be covered with a transparent glass or plastic sheet to prevent contamination.

Economic media are used to culture spirulina

Liquid effluents taken from well digested human excreta,

modified sea water and Zarrouk medium are used for this purpose

Zarrouk’s Medium for Indoor Cultures

Macronutrients (g/l)

• Sodium bicarbonate (18)• Potassium hydrogen Phosphate (0.5)• Sodium carbonate (4)• Sodium chloride (1)• Sodium nitrate (2.5)• Magnesium sulphate (0.2)• Ferrous sulphate (0.01)• Potassium sulphate (1)• Calcium chloride (0.04)• EDTA (0.08)

Micronutrients(In Traces)• Magnesium Chloride• Sodium Molybdate• Zinc Sulphate• Sodium tungstate• Titanium Sulphate• Cobalt Nitrate

• Human excreta is diluted and digested in a digester at 55˚c for 10 days.

• Liquid effluent is then taken from the digester and filtered through a fine cloth to get a filtrate.

• The filtrate is again filterd through a sand bed filter to remove the contaminants.

• The final filtrate is used as a medium for spirulina culture.

• Sea water is an another economic medium.• The sea water is treated with a NaHCO3 to

precipitate out excess calcium and magnesium ions.

• To this water, K2HPO4, FeSO4 and urea are added to make it a culture medium.

• The pH of the culture media is adjusted to 8.5.

• A medium is filled in the open circulating tank and the tank is inoculated with a small volume of pure spirulina culture.

• The culture is stirred continuously for giving aeration to it.

• The culture is illuminated with a low intensity light, if it is inside the room.the temperature is maintained at 35-40˚c.

• Spirulina grows rapidly and forms a bloom in the culture.

OXIDATION POND SYSTEM

• In this method all solid wastes and suspended particles are removed from the sewage by primary and secondary treatments.

• The sewage water is allowed to flow into an oxidation pond.

• A few liters of culture is inoculated into the pond as a starter culture .

• Spirulina grows in the natural system and produces a dense mat on the surface of the sewage water

Open Pond

Texcoco lake

Natural Open Pond

Texcoco lake

2. Closed SystemPhotobioreactor

• A photobioreactor is a bioreactor which incorporates some type of light source. Virtually any translucent container could be called a photobioreactor, however the term is more commonly used to define a closed system, as opposed to an open tank or pond.

• A pond covered with a greenhouse could be considered a photobioreactor.

• A photobioreactor can be operated in "batch mode" but it is also possible to introduce a continuous stream of sterilized water containing nutrients, air, and carbon dioxide.

Various models ofPhotobioreactor

Green-house

Indoor Culture

Requirements for Growth of Spirulina

(i) Algal tanks. Generally, circular or rectangular cemented tanks are constructed. The circular tanks are more preferred over the rectangular one because of ease in handling. Size may be according to convenience and yield needed. Depth should be about 25cm.

Open tanks are suitable for tropical and subtropical regions.

(ii) Light. Low light intensity is required at the beginning to avoid photolysis. Spirulina exposed to high light intensity is lysed.

(iii) Temperature. Temperature for optimum growth should be between 35-40°C.

(iv) pH. Spirulina grows at high pH ranging from 8.5 to 10.5. Initially, culture should be maintained at pH 8.5 which automatically is elevated to 10.5.

Factors Affecting Biomass Production

• (i) Illumination time;• (ii) Light intensity;• (iii) Supply of CO2. Concentrations of CO2 differ in different

conditions, for example, an alkaline lake. Lake Texcoco in Mexico, has high concentration of sodium carbonate. On the other hand, algal growth is limited as a result of liberation of CO2 and ammonia by bacterial activity;

• (iv) Nitrogen sources (ammonium salts or nitrates are the suitable nitrogen sources which increase biomass yield);

• (v) Agitation of growing cells to maintain cells in suspension.

Harvest of Spirulina Biomass

• Spirulina filaments develops gas vacuoles in the cells and hence they float on the surface of culture medium,

• As the density increases, a dense bluish green mat develops on the surface of the culture,

• The biomass is harvested by filteration using a fine mesh or cloth.

• In this way 12-20gm of spirulina/m2/day can be harvested from a pond.

DRYING AND PACKING• The harvested biomass is washed with an acid water(ph4) to

remove toxic pollutants.• Then, it is washed with tap water to remove the acidity of the acid

water.• The washed biomass is spread on a polythene sheet in the

sunlight.• On drying thin flakes of biomass develops on the plastic sheet.• The dried flakes are collected and made into powder.• The dried biomass is packed in aluminium lined bags or sealed

bottles for marketing.• Spray drying is followed in the dark, if the biomass has to be used

to make pills along with a vit A and C.

USES OF SPIRULINA SCP

SPIRULINA AS HEALTH FOOD Rich source of proteins, vitamins, and

minerals. Its digestibility is a high as 84%. Eating 20gm of spirulina scp can fulfil the

essential amino acids, vitamins, and minerals

• SPIRULINA AS THE THERAPEUTIC AGENT

• To reduce Body weight,Choloestrol.Blood sugar level in diabetics.• Promotes wound healing.• Taking a 3gm of spirulina in the previous night reduces pre-

menstrual stress in women.• Phycocyanin present stimulates the immune system to provide

resistance to the body against pathogenic diseases.• Act as a anticancer agent to prevent cancer risks(beta carotene)• Increases lactation in nourishing mothers• Increases the eye power via synthesis of vitamin A

• It is a rich source of Gamma-Linolenic Acid (GLA) a rare essential fatty acid.

• It is world's richest natural source of Vitamin B-12 and Pro-Vitamin 'A'.

• It is best natural source of anti oxidants, a natural anti -ageing drug.

• it contains a whole spectrum of natural mixed carotene and xanthophyll phytopigments.

• Spirulina is 100% natural, nontoxic and cholesterol-free. There are No known side effects of spirulina.

• It is consumed in its most natural form as no artificial flavours or colours are added in Spirulina products.

• Researchers at NASA found it to be an excellent, compact space food for astronauts, as it is believed that 1 kg. of Spirulina is equivalent to 1000 kgs of assorted vegetables.

• International health organizations have also summoned Spirulina as one of the "Greatest Super foods on Earth".

• Spirulina is being developed as the "food of the future" because of its amazing ability to synthesize high-quality concentrated food more efficiently than any other algae.

It repairs body tissues and provides immunity and thus strengthens body's defense mechanism.

It enhances the body's ability to generate new blood cells.

In cosmetics

• Spirulina contains high quality of proteins and vitamin A and B. These play a key role in maintaining healthy hair.

• Many herbal cosmeticians are making efforts to develop a variety of beauty products. Phycocyanin pigment has helped in formulating biolipstics and herbal face cream in Japan. These products can replace the present coaltar-dye based cosmetics which are known as carcinogenic.

Spirulina capsules

Spirulina flakes

Spirulina tablets

Spirulina powder

spirulina DOSAGE

• Spirulina dosage depends largely on the form being taken. Spirulina products come in the form of spirulina capsules, tablets, powder and flakes. Spirulina extract is also available for those who prefer this type of spirulina diet.

Spirulina may also be found in combination with other natural products such as chlorella, wheat or barley grams.

• About ten (to twenty) 500 milligram spirulina tablets a day for adults and five (to ten) for children under 12 are usually recommended.

• The dose for spirulina capsules and tablets can vary from 250 milligrams to 5 grams a day as well. About a spoonful of spirulina extract or spirulina powder (about 3 to 5 grams) is enough to last a day.

Spirulina OVERDOSE

• Spirulina is a completely natural food source. Being natural, it is not expected to give any problems to the body.

• Theoretically speaking, even if too much is taken, there should be no harm done to the body.

ADVERSE EFFECTS OF Spirulina

• Some people have reported to experience some minor symptoms after taking spirulina. These spirulina side effects include:

• Slight fever. This can be normal since the body needs to burn extra protein found in spirulina.

• Slight dizziness and nausea. Thirst and constipation. When taking spirulina, you should increase your water intake. At least half a liter should be taken for the body to absorb the spirulina herb well.

• Stomach ache and other gastrointestinal symptoms Slight body risk or skin itching

Danger of Spirulina

• Spirulina poses no dangers when taken in the recommended dosage.

• Taking less than 50 grams per day should be relatively safe with no notable side effects. However, as in most dietary supplements, taking exceedingly large doses can cause unwanted spirulina side effects.

• Because of the high nucleic acid content of spirulina, it can give you increased levels of uric acid, which can consequently cause kidney problems.

• Due to the high content of vitamins and minerals, impaired liver function might also occur.

Single cell proteins from Chlorella

• Chlorella is general nature and single-cell green algae

• It is a single-cell and freshwater algae. Its spherical diameter is 3 to 8 micron.

• It is a plant with high efficient photosynthesis and has a wide range of distribution.

• Its growth depends on photosynthesis. • its cells are rich in chlorophyll, so its photosynthesis

is very strong. Its rich contents of protein, vitamins, minerals, dietary fiber, nucleic acid.

The mass culture of chlorella

• Types of culture: 1. Closed Sterile Culture 2. Open semi-sterile cultures 3 .Open phycomycete polyculture

1. Closed Sterile Culture

• using reagent grade chemicals, purified water, added EDTA and vitamins, and other nutrients to produce senior medical products, fine chemicals, and advanced oils.

2. Open semi-sterile cultures

• using pure chemicals and purified water to produce algae powder, health food, pigment and pharmaceutical products.

3 .Open phycomycete polyculture

• using Vegetables, meat processing and the clean organic liquid waste of fermentation and industrial distillation as culture medium to produce algae powder, health food, pigment and pharmaceutical products

conditions of Chlorella large-scale cultivating

1.Climate is a top priority issue to be considered for large-scale cultivation.

The growth of large-scale cultivation has nothing to do with culture approach, but depends on the illumination area and the size of earth's surface of the solar radiation.

Using artificial light and heating measures could control climatic conditions. Because of high costs, the practical application is uneconomical in the economy.

Tropical regions have the ideal temperature.

2 .The addition of CO2• According to the research that Chlorella

accumulating 1 mg of dry matter needed 1 mol (1.8mg) CO2 by photosynthesis.

• The use of CO2 (0.04 mol / L) dissolved in water is far from being able to meet the needs of Chlorella Growth.

• The addition of CO2 is an effective way to increase production for large-scale cultivation.

3. stirring:• Due to the variety of factors, the algal cells are often

vulnerable to be impact by the precipitation and concentration in the large-scale cultivation, which seriously affects the reproduction and growth rate. As a result, the growth of algae cells may appears no balance.

• Therefore, it is necessary to design stirring in large-scale cultivation.

• Stirring four to five times a day can increase air and carbon dioxide contacting with the algal cells.

Harvest of algae cells

⑴Precipitation method: using alum or Limewater to make the algal cells agglutinate and precipitate, but the approach seriously affect the quality and food value of the products;

⑵Filtration method: as the Chlorella cells is only 1 to 5

microns, it is not suitable for filtration; ⑶Centrifugation method: This method is applicable to the

separation of Chlorella. The quality of product is good, but the cost is too high.

.

Drying of algal cells

Dry powder is the primary product of chlorella of large-scale cultivation.

the time of long drying leads to chlorophyll losing and affects the product quality. The following are the usable methods:

⑴Solar drying: the method is simple. Products are good quality. But the drying quantity is less and impact by the weather;

⑵Drum drying: as Chlorella with serious visco-wall, this method is not suitable;

⑶Spray drying: high yield, high digestibility of algae powder, good quality;

⑷Freeze-drying: applying to the production of medical supplies and bioactive preparation.

Factors affecting the growth of Chlorella

1. Light: Light energy conversion rate of converting Chlorella into cell substances is 2.6 ~ 7.1%,which is equivalent to 1.9 to 1.9～ 4.9 g*m-2*d-1 of dry matter cells biomass.

• Illumination intensity relates to the density of culture in large-scale cultivation.

• If the density of culture increases, the light intensity decreases, as a result the growth will drop.

• For large-scale cultivation, the temperature difference between the day and night is one of the necessary factors to maintain the high yield.

2 .magnetic fields: Different magnetic induction intensity has different impacts of Chlorella growth and nutrients. Weak magnetic induction treatment stimulates the growth of Chlorella. Strong magnetic induction intensity more than 0.05 T restricts its growth.

• After weak Magnetic dose treatment, the sugar and protein content of chlorella increase, but its fat content slightly decreases;

• after strong magnetic dose treatment, the sugar and protein content of chlorella decrease, but its fat content increases significantly.

• Magnetic treatment changes the composition of amino acids. The amino acid content of chlorella has increased after magnetic treatment.

Cultivation of algae in india

• Spirulina platensis is grown at Delhi (Indian Agricultural Research Institute) and

• Nagpur (National Environmental Engineering Research Institute),

• Chlorella is being grown at Pondicherry (Auroville Centre for Environmental Studies).

Spirulina Production Unit at the Indian Agricultural Research Institute, New Delhi

Conclusion

At present, production of SCP by mass culture of micro-organisms is in its infancy. It needs much boost to solve the problem of starvation in coming decades.

One of the ways to enhance productivity and quality of SCP product is the genetic improvement of micro-organisms.

Moreover,transfer and expression of beneficial genes in the micro-organisms have opened a new era for the organisms of algal proteins and other compounds to used in food and feed .

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