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SINGLE CELL PROTEINS

ITS PRODUCTION
From Chlorella and Spirulina

A Powerpoint presentation
By: 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 B.VITAMINS
• Crude proteins 65% • Biotin 0.2%
• Carbohydrate 16% • Cyanocobalamin 66.0mg
• Lipids 6.7% • Folic acid 18.0mg
• Nucleic acids 4.2% • Riboflavin 1.8mg
• Thiamin 0.12mg
• Tocopheol 0.81IU
• β-carotene 320,000IU
MINERALS ESSENTIAL AMINO ACIDS
• Calcium 6.6mg • Lysine 2.99%
• Phosphorus 0.9gm • Cystine 0.47%
• Iron 4mg • Methionine 1.38%
• Sodium 0.8mg • Phenylalanine 2.87%
• Potassium 1.3mg • Threonine 3.04%
MASS CULTURE OF SPIRULINA
Spirulina is cultured in large scales in artificial
ponds or tanks and oxidation ponds.
• Open Culture
1. open circulating system
2. oxidation pond system
• Closed Culture
1. Photobioreactor
2. 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) Micronutrients(In Traces)


• Magnesium Chloride
• Sodium bicarbonate (18) • Sodium Molybdate
• Potassium hydrogen Phosphate (0.5)

• Zinc Sulphate
Sodium carbonate (4)
• Sodium chloride (1) • Sodium tungstate
• Sodium nitrate (2.5) • Titanium Sulphate
• Magnesium sulphate (0.2) • Cobalt Nitrate
• Ferrous sulphate (0.01)
• Potassium sulphate (1)
• Calcium chloride (0.04)
• EDTA (0.08)
• 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
Natural Open Pond

Texcoco lake
Texcoco lake
2. Closed System
Photobioreactor
• 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 of
Photobioreactor
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 .
THANK YOU

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