SINGLE-CELL

PROTEIN
SINGLE-CELL
PROTIEN
 What is Protein ???
Any of a group of complex organic macromolecules that contain carbon,
hydrogen, oxygen, nitrogen, and usually sulphur and are composed of one
or more chains of amino acids.
Proteins are fundamental components of all living cells and include many
substances, such as enzymes, hormones, and antibodies, that are
necessary for the proper functioning of an organism.
They are essential in the diet of animals for the growth and repair of
tissue and can be obtained from foods such as meat, fish, eggs, milk, and
legumes.
 Protein requirement:
In inactive lifestyle, the recommended dietary allowance (RDA) for a sedentary
individual is
about 0.8 grams per kilogram of body .

So, if weight of body is 90 kg then protein requirement

90 x 0.8 = 72g per day protein is required.

If lifestyle is active or hard working then protein required is 1.4g/kg to 1.8 g/kg
of body weight daily.
 Daily requirements (g) of essential aminoacids for the
human adult Data retrieved from FAO ([Link]

Essentials amino FAO

acids recommendation Minimum
Phenylalanine 2.2 1.1
Methionine 2.2 1.1
Leucine 2.2 1.1
Valine 1.6 0.8
Lysine 1.6 0.8
Isoleucine 1.4 0.7
Threonine 1.0 0.5
Tryptophan 0.5 0.25
Total 12.7 6.35
 WHY DO WE NEED ALTERNATIVE SOURCES OF
FOOD?

 About 50 years ago (1934-1938) the less developed areas of

the world were the main exporters of grain to the developed
world.

 Since 1948 the food flow has reversed, from the developed
world to the less developed, mainly due to the rate of growth of
the world's population which was much higher in the less
developed countries.

 This means that during the 35-year period (1980-2015) we

must produce as much food as we have since the dawn of
agriculture about 12000 years ago.
 REALITY
Death from starvation, malnutrition and related
diseases is a reality in many countries today.
The World Health Organisation (WHO) estimates
that 12,000,000 people die of hunger and starvation
related diseases every year. Half are children under
the age of 5.
• The increasing world deficiency of protein is becoming a main problem of
humankind.
• Since the early fifties, intense efforts have been made to explore new,
alternate and unconventional protein.
• For this reason, in 1996, new sources mainly yeast, fungi, bacteria and algae
named Single Cell Protein (SCP) as coined to describe the protein production
from biomass, originating from different microbial sources.
• Microbial biomass has been considered an alternative to conventional
sources of food or feed.
 Large-scale processes for SCP production show interesting features,
including:
• The wide variety of methodologies, raw materials and microorganisms
that can be used for this purpose
• High efficiency in substrate conversion
• High productivity, derived from the fast growth rate of microorganisms
• Independence of seasonal factors
 History
A survey of the history of the use of microorganisms for human
consumption indicates three major trends:

(a) microbes as a source of enzymes in the food industry (baking,

brewing, distilling, wine making, cheese production) has at
present a new aspect-the use of immobilized enzymes of microbial
origin,
(b) microbes as producers of nutritive substances (amino acids,
nucleotides, vitamins, organic acids, sugars, aromatizers) and
(c) direct utilization of microbial biomass as foodstuff.
 HISTORY
• Used to named as Microbial Proteins.
• 1966: M.I.T Professor Carroll [Link] renamed it as “Single Cell
Proteins”
• Transportation of food was common in the past but with the
increase in population, energy crisis has encounter the world.
• In 60s’, idea that the dried cells of micro-organisms can become
an ultimate part to solve this problem.
• Thus gained research interest among the scientists & industries
(specifically oil industry).
• In 20th Century, the SCP technology for the
production of protein-rich contents from
the microbes on the large scale was
established.

• 1950s: Food-from-oil.

• 1960s: British Petroleum Industry

developed technique named as “Protein-
from-oil Process” using yeast fed on waxy
paraffin, a product of oil refineries.
ADVANTAGES OF USING MICROORGANISMS

1. MO grow at very fast rate under optimal conditions

2. Quality and quantity is better than higher plants and
animals
3. Wide range of raw materials can be used
4. Culture and fermentation conditions are simple
5. MO can be genetically manipulated
Advantages of SCP over conventional protein sources
are:
It has high protein and low fat content.
It is good source of vitamins particularly B-complex. e.g. Yeasts
It can be produced through-out the year.
Waste materials are used as substrate for the production of these
proteins.
It reduces the environmental pollution and helps in recycling
of materials.
SCP organisms grow faster and produce large quantities of SCP from
relatively small area of land and time.
These have proteins with required amino acids that can be easily
selected by genetic engineering.
During the production of SCP biomass, some organisms produce useful
by products such as organic acids and fats.
It can be genetically controlled.
It causes less pollution.
Algal culture can be done in space which is normally unused.
Factors that impairs the usefulness of Unicellular biomass

All single-celled microorganisms of interest from the industrial

point of view have a nondigestible envelope, which makes protein
assimilation difficult.
The content of nucleic acids in the unicellular biomass is higher
than the permissible level and may cause disorders of purine
metabolism in the human body.
The biomasses of some unicellular microorganisms have an
unpleasant color (algae), taste, and smell, which make them
unsuitable even for animal consumption.
Food grade production of SCP is more expensive than other
sources of proteins, as it depends on the raw [Link]
for human consumption is 10-12 times more expensive than
SCP for animal feed.
Digestion of microbial cells is rather slow, and is frequently
associated with indigestion and allergy reactions.
SINGLE CELL PROTEINS
SINGLE CELL PROTEINS

• The term “Single Cell Protein” refers to the total

protein extracted from the pure cultures of
microorganisms (e.g. yeast, algae, filamentous
fungi, bacteria) and can be used as a protein-rich
food supplements by humans and animals.

• Also known as ”Microbial Protein”

• Single-cell proteins develop when microbes ferment waste materials
(including wood, straw, cannery, and food-processing wastes,
residues from alcohol production, hydrocarbons, or human and
animal excreta).

• 60-80% dry cell weight; contains nucleic acids, fats, CHO, vitamins and
minerals Rich in essential amino acids (Lys-Met)

• The problem with extracting single-cell proteins from the wastes is

the dilution and cost.

• Found in very low concentrations, usually less than 5% .

• Engineers have developed ways to increase the concentrations

including centrifugation, flotation, precipitation, coagulation, and
filtration, or the use of semi-permeable membranes.
 Microorganisms

Bacteria
Methylophilus methylotrophus
Pseudomonas sp.
Brevibacterium sp.
Mushrooms
Yeasts Agaricus
Lactobacillus bulgaricus Morchella
Candida lipolytica Vovariella
Bakers yeast
Kluyveromyces fragilis

Fungi
Trichoderma viridae
Aspergillus niger

Actinomycetes
Nocardia sp
Thermomonospora fusca
Algae
Chlorella and
Spirulina
 “Possible Substrates for SCP”

They can be subdivided into three categories:

 High energy sources (natural gas, n-alkanes, gas-
oil, methanol, ethanol, acetic acid);
 Different wastes (molasses, sulfite waste liquor,
milk, whey, fruit wastes); and
Renewable plant resources (sugar, starch,
cellulose).
A wide range of substrates can be used to grow
microbial proteins

• whey, orange peel residue, sweet orange residue, sugarcane bagasse,

paper mill waste, rice husks, wheat straw residue, cassava waste, sugar
beet pulp, coconut waste, yam waste, banana pulp, mango waste,
grape waste, sweet potato
Average composition of the main groups of micro-organisms
(% dry weight)

TABLE : 1

Fungi Algae Yeasts Bacteria

Protein 30-45 40-60 45-55 50-65
Fat 2-8 7-20 2-6 1.5-3.0
Ash 9-14 8-10 5-9.5 3-7
Nucleic 7-10 3-8 6-12 8-12
Acids
1. Yeasts and Fungi
Filamentous Fungi used for SCP production are Chaetomium celluloliticum,
Fusarium graminearum, Paecilomyces varioti which grows on cellulose waste,
starch, and sulphite waste liquor respectively and content about 30 – 55 %
protein.

SCP is produced from yeasts viz. Candida utilis, Candida lipolytica, Saccharomyces
cervicea.
Torula yeast (which grows on Ethanol) as a food is obtained through fermentation
using molasses as substrate and it has high protein–carbohydrate ratio than
forages. It is rich in lysine but poor in methionine and cysteine. Saccharomyces
consists of high protein with good balance of amino acids and rich in B–complex
vitamins. It is more suitable as poultry feed.
Yeast are higher in lysine content.
Strict aseptic conditions are required when using Yeast as a SCP production.

Disadvantages:
High nucleic acid content.
Slow growth is observed in Fungi vis-à-vis than yeast & bacteria.
Contamination risk.
Mycotoxins are also produced.
2. BACTERIA
• They have more than 60% protein but are poor in sulphur containing
amino acids.
• Brevibacterium uses hydrocarbons while Methylophilus methylitropous
uses methanol as a substrate.
Disadvantages:
• It has high nucleic acid content.
• Recovering the cells is a bit problematic.
• Endotoxin production should be carefully tested.
3. ALGAE:
• Chlorella, Scenedesmus acutus and Spirulina maxima are grown for
SCP.
• These have about 60% protein with good amino acid composition but
less in sulphur containing amino acids.
• Chlorella and Spirulina are used for commercial scale production in
Taiwan, Thailand, Japan, Israel, Mexico and USA.
• It is spray dried and sold as pills and powders.

Disadvantages:
• As they are rich in Chlorophyll, it is not advised for human
consumption (except Spirulina).
• It has low density.
• There is lot of risk of contamination during growth.
Microorganism Substrate Used as Used commercially
Algae
Chlorella sp. CO2 + sunlight Feed Yes (Japan and Taiwan)
Scenedesmus acutus CO2 + sunlight -
Spirulina maxima CO2 + sunlight Feed Yes (Mexico)
Yeasts
Candida utilis (Torula Yeast) 1. Confectionery - Yes (U.K.), Symba process
2. Ethanol Feed Yes (USA)
3. Sulphite liquor - Yes (Europe, USA, Russia)
C. intermedia Whey - Yes; Vienna process
C. krusei (+ Lactobacillus
Whey - Yes; Kiel process
bulgarius)
n-alkanes (C10 - C23) +
C. lipolytica - Yes (Russia)
ammonia
Kluyveromyces fragilis Whey Food Yes (France); Le Bel process
Saccharomyces cerevisiae Molasses (Food)* Yes
Fungi
Chaetomium cellulolyticum Cellulosic wastes - Promising
Fusarium graminearum Starch hydrolysate Yes (U.K)
Paecilomyces varioti Sulphite liquor - Yes (Finland); Pekilo process
Bacteria
Brevibacterium sp. C1 - C4 hydrocarbons - Process developed
Methylophilus
Methanol Feed Yes (U.K.),
methylotrophus
Properties of SCP
One of the main advantages of SCP compared to other types of protein
is the small doubling time of cells (td) as shown in Table

TABLE : 2

Organism Mass Doubling

Bacteria and yeast 10-120 min

Mold and Algae 2-6h

Grass and some plants 1-2wk

Chickens 2-4 wk

Pigs 4-6 wk

Cattle 1-2 mo

People 0.2-0.5 yrs

Due to this property, the productivity of protein from micro-organisms is greater
than that of traditional proteins

Efficiency of protein production of several protein sources in 24 hours ( 16)

TABLE : 3

Organism Amount of Protein

(1,000 kg)
Beef Cattle 1.0 kg
Soybeans 10.0 kg
Yeast 100.0 tn
Bacteria 100x10,000,000 tn
Average composition of the main groups of micro-organisms
(% dry weight)

TABLE : 4

Fungi Algae Yeasts Bacteria

Protein 30-45 40-60 45-55 50-65
Fat 2-8 7-20 2-6 1.5-3.0
Ash 9-14 8-10 5-9.5 3-7
Nucleic 7-10 3-8 6-12 8-12
Acids
 Bacterial protein is similar to fish protein, yeast's protein
resembles soya and the fungi protein is somewhat lower than
the yeast's. Of course microbiological proteins are deficient
in the sulphur amino acids cysteine and methionine and
require supplementation, while they exhibit better levels of
lysine.
 Essential amino acid content of the cell protein in
TABLE : 4 comparison with several reference proteins
(grams of amino acid per 100 grams of protein)

Spirulina maxima
Amino Acids

Saccharomyces
Cellulomonas

PeniciIllium
cerevicia

notatum

Wheat

Cow
milk
Egg
Lysine 7.6 7.7 4.6 3.9 2.8 6.3 7.8

Threonine 5.4 4.8 4.6 - 2.9 5.0 4.6

Methionine 2.0 1.7 1.4 1 1.5 3.2 2.4

Cysteine - - 0.4 - 2.5 2.4 -

Tryptophan - 1.0 1.4 1.25 1.1 1.6 -

Isoleucine 5.3 4.6 6.0 3.2 3.3 6.8 6.4

Valine 6.5 3.9 5.4 3.9 4.4 7.4 6.9

 The vitamins of micro-organisms are primarily of the B type, B12 occurs mostly in
bacteria, while vitamin A is usually found in algae. Table shows the vitamin content
of various food MO; Vitamin content of various food micro-organisms (mg/100 g
dry weight)
Other nutritional parameters which evaluate the quality of a given SCP are:
- the digestibility (D)
- the biological value (BV)
- the protein efficiency ratio (PER)
- the net protein utilisation (NPU)
 The quality of SCP is an important factor for commercial
production.
1. First parameter which reflects the quality of a protein, is
the is the percentage of the total nitrogen Digestibility
Coefficient (DC) consumed which is absorbed from the
digestive tract.
2. Estimation of the Biological Value (BV) is a measure of
nitrogen retained for growth or maintenance.
3. An accurate method to evaluate the quality of proteins
is the determination of the Protein Efficiency Ratio (PER),
expressed in terms of weight gain per unit of protein
consumed by the test animal in short-term feeding trials.
4. Finally, the Net Protein Utilization (NPU) -equivalent to
the calculation BVxDC-is a measure of the digestibility of
the protein and the biological value of the amino acids
absorbed from the food.
Nitrogen and Protein contents of microbial cells compared
with selected foods of animal and plant origin
TABLE : 5

Source Nitrogen Crude Protein

(%) (%)
Filamentous fungi 5-8 31-50
Algae 7.5-10 47-63
Yeast 7.5-8.5 47-53
Bacteria 11.5-12.5 72-78
Milk 3.5-4.0 22-25
Beef 13-14.4 81-90
Egg 5.6 35
Rice 1.2-1.4 7.5-9.0
Wheat Flour 1.6-2.2 9.8-13.5
Corn meal 1.1-1.5 7.0-9.4
 Problem of Nucleic Acids
About 70-80% of the total cell nitrogen is represented
by amino acids while the rest occurs in nucleic acids.
This concentration of nucleic acids is higher than other
conventional proteins and is characteristic of all fast
growing organisms.
The problem which occurs from the consumption of
proteins with high concentration of nucleic acids (78
g/100 g protein dry weight) is the high level of uric acid in
the blood, sometimes resulting in the disease gout.
Uric acid is a product of purine metabolism.
 Problem of Nucleic Acids
 Most mammals, reptiles and molluscs possess the
enzyme uricase, and the end product of purine metabolism
is allantoin.
 Man, birds and some reptiles lack the enzyme uricase
and the end product of purine degradation is uric acid.
 The removal or reduction of nucleic acid content of
various SCP's is achieved with one of the following
treatments: chemical treatment with NaOH; treatment of
cells with 10% NaCl; thermal shock.
 These methods aim to reduce the RNA content from
about 7% to 1% which is considered within acceptable
levels.
Production Of SCP:

•Production of SCP involves following steps:

1. Selection of Strain of microbe and Substrate
2. Fermentation
3. Harvesting
4. Post harvest treatment
5. Processing of SCP
Basic Steps of SCP production:
Selection of Strain of Microbe
& Substrate:
• Very Crucial step.
• Microbe selected shouldn’t produce toxicity
in its biomass.
• It should not be harmful for a consumer to
consume.
• Selected microbe should produce a large
quantity of protein.
• Substrate should be cheap, effective, allow
favorable growth and ease of isolation.
Fermentation:

• Is done in a large chamber either of glass or

stainless steel called “Fermentor”.
• Fermentation should be done under
sterilized conditions.
• Controlled conditions as necessary e.g.
Temperature, Pressure, pH, Humidity etc.
• Usually fed-batch cultures are used for the
fermentation of microbes.
General process for production of SCP
 Harvesting
• For the producing and harvesting of microbial proteins cost is a major
problem.
• There are many methods available for concentrating the solutions like
filtration, precipitation, centrifugation and the use of semi-permeable
membranes.
• The equipment used for these methods of de-watering is expensive and
so would not be suitable for small scale productions and operations.
• Single cell proteins need to be dried to 10% moisture or they can be
condensed and denatured to prevent spoilage.
 Biomass Harvesting
• The microbial biomass can be harvested by a variety of methods.
• Single cell organisms like yeast and bacteria are normally
recovered by centrifugation, flocculation and floatation.
• Filamentous organisms are recovered by filtration. It is important
to recover as much water as possible prior to final drying.
• The whole operation is to be done under clean and hygenic
conditions to keep the product and the broth that leaves the
plant free of bacterial contamination.
• In some cases, an after-treatment of the biomass is desirable to
reduce the unwanted compounds in the product or to isolate the
protein.
• One of the important tasks is to reduce nucleic acid content,
which is high in microorganisms (4-6% in algae, 10-16% in
bacteria, 6-10% in yeasts and 2.5-6% in fungi) and can be
hazardous to health.
Post-harvesting Fermentation:
• Isolated microbial colonies are subjected to various differential
techniques.
• E.g. Centrifugation, Washing, Drying etc.
• Produced protein contain impurities in it e.g. carbohydrates,
nucleic acids, lipid contents, salts etc
• Pure protein isolation can be done by disrupting the cell wall
through crushing, crumbling, cycles of freezing & thawing,
grinding & thermal shocks.
• Nucleic acid can be remove by:
1. By treatment with Nacl 10%
2. By Chemicals e.g. NaoH
3. Thermal shocks
4. Enzymes Treatment e.g. ribonucleases
Raw material Organism Scale Product Organization

Food yeast Universal Foods

Cheese whey Kluyveromyces fragilis 5000 tons/year fermentation nutrient Corporation,
Juneau, Wisconsin

10,000 Torula yeast, food Pure culture

Ethanol Candida utilis tons/year ingredient products, Hutchinson,
Minnesota

20,000 to All Union Research

Institute of Protein
N-paraffins Candida guillienmondis 40,000 Food yeast Biosynthesis,
tons/year USSR

Rhinelender paper
Sulfite waste Candida utilis 15 tons/year Torula yeast corporation
liquor Rhinelender
Wisconsin
 Raw material Organism Scale Product Organization

Glucose (Food 50-100 Mycoprotein Ranks Hovis Mc Dougall,

Grade) Fusarium graminearium tons/year (human food) High Wycombe,UK

Cheese whey Penicillium cyclopium 300 tons/year Animal feed Heurty , S.A.,France

Coffee waste Trichoderma harzianum 40,000 lit Animal feed ICAITI,Guatemala and El
Salvador

10,000 Animal Tampela and Finnish Pulp

Sulfite waste and paper Research
liquor Paecilomyces varioti tons/year feed(Pekilo Institute,Jamsankoski,
protein) Finland

1 tons/day Animal Envirocon Ltd,Vancouver,

Pulp mill wastes Chaetomium cellulyticum feed(Waterloo BC, Canada;University of
process) Waterloo,Ontario
 Organism Raw material Production Producer or developer

2 metric Taiwan Chlorella

Chlorella sp. CO2 tons/day Manufacture [Link], Taipei

20 Central Food Technological

Scenedesmus CO2,Urea grams/ Research
acutus square Institute,Mysore,India
meter/day

CO2, or
Spirulina NaHCO3, 320 metric Sosa Texococo,SA Mexico
maxima Na2CO3 tons/year City
Economic parameters : The Scope of SCP production
i. Products obtained via microbiological synthesis must be
competitive with traditional food sources.
ii. When estimating costs involved in SCP production, such major
factors as the biomass yield, cooling, and oxygen requirements
should be taken into consideration.
iii. They depend not only on the choice of substrate, but on the choice
of the microorganism as well. All this determines the cost of
production and economic feasibility.
iv. It is obvious that one of the major factors limiting the use of
hydrocarbon yeast is the residual hydrocarbon content.
v. Demands of the country or its separate regions for protein of a
particular type
vi. Expenditures for the delivery of finished products to the places of
their consumption
[Link] of by products.
 ACCEPTABILITY OF SCP AS A HUMAN FOOD AND ITS
TOXICOLOGY CONCERN

• Raw materials used in production of SCP are the main safety hazard.
•The acceptability of SCP when presented as a human food does not
depend only on its safety and nutritional value but also the mind
frame of people to consume material derived from microbes which is
concerned to social and ethical issues like psychological, sociological
and religious implications.
•A more intensive clinical and toxicology testing including short-term
acute toxicity testing (animal species) and followed by extensive and
detailed long term studies.
•And in return incurs a huge scientific and financial investment.
Biolipsticks and herbal face creams are produced in Japan
from the phycocyanin pigment of Spirulina.

Formulation
of SCP
 APPLICATIONS
1. As protein supplemented food-
• Also source of vitamins, amino acids,
minerals, crude fibers, etc.
• Supplemented food for undernourished
children.

2. As health food-
• Controls obesity
• Provides instant energy .
• Example- Spirulina- part of diet of US
Olympic team.
3. In therapeutic and natural
medicines-
• Reduce body weight, cholesterol,
stress.
• Lowers blood sugar level in
diabetic(due to presence of B -
linolenic acid)
• Prevents accumulation of cholesterol
in body.
• Healthy eyes and skin (beta carotene)
• Beta carotene ( anti cancer substance-
UN National Cancer Research
Institute)
• Increase lactation.
 MEDICINAL USES OF SPIRULINA
• Strengthen and improve immune system
• Phycocyanins build blood cells
• Increase antiviral activity
• Exhibits anti cancer activity
• The US Olympic teams take spirulina tablet as a source
of instant energy.
 Studies showed that spirulina consumption of 4 weeks
reduced serum cholestrol level in human beings by 4.5%
and significantly reduced body weight by 1.4±0.4 kg
after 4 weeks.
 There is no changes in clinical parameters (Blood
Pressure) or in biochemical variables (haemoglobin,
blood cells, sedimentation rate) and absence of adverse
4. In cosmetics-
• Important role in maintaining healthy
hair (vitamin A and B).
• Many herbal beauty products.
• Biolipstics and herbal face
cream(Phycocyanin).
• Capable of replacing coal tar dye based
cosmetics.

5. Poultry and cattle feed-

• Excellent, convenient source of protein
and other nutrients.
• Used to feed cattle, fishes etc.

6. In the technical field as:

• Paper processing, leather processing and
as foam stabilizers .
CONCLUSION

• At present SCP production is in its infancy. One of the ways to

enhance productivity and quality is genetic improvements of
micro-organisms.
• Using microbial biomass as a food source deserves serious
consideration because of insufficient world food supply and high
protein content of most micro-organisms.
 For future
success of SCP
First,
food technology problems have to be
solved in order to make it similar to
familiar foods and
Second,
the production should compare
favourably with other protein sources.
Sample of the single cell protein
biosolids after the drum dryer.
 THANK YOU
-PHARMA STREET