Single Cell Protein 1

Single Cell Protein

(SCP)

MBB 603: Advances in Microbial Biotechnology

Submitted to:
Dr. Chintan Kapadiya
Assistant professor
Dept. of Plant Mol. & Biotech.
A.C.H.F., N.A.U.
Navsari-396 450
Submitted by:
Patel Hiren K.
Ph.D. (Sem-6)
Dept. of Plant Mol. & Biotech.
N.M.C.A., N.A.U.
Navsari-396 450

Hiren K. Patel 2014

 Single Cell Protein 2

Single-cell protein (SCP)

Definition:
Single-cell protein (SCP) typically refers to sources of mixed protein extracted from pure or
mixed cultures of algae, yeasts, fungi or bacteria (grown on agricultural wastes) used as a
substitute for protein-rich foods, in human and animal feeds.

Introduction:
Most of the developing countries of the world are facing a major problem of malnutrition.
Due to rapid growth in the population deficiency of protein and nutrients are seen in human
food and as well as animal feed. It has been estimated that if necessary measures are not
taken the malnutrition condition will lead to some major crisis in the developing countries.
Therefore it is very important to increase protein production and also its availability to the
population by utilizing all the available ways and also methods. The increased world demand
for food and in particular feed protein spurred the search for non-conventional protein
sources to supplement the available protein source.

The need for large scale production of single cell protein is the need for more protein in the
diet of humans. Single cell protein typically refers to proteins extracted from pure culture or
mixed culture of microorganisms such as algae, yeasts, fungi or bacteria. These extracted
protein are used as a substitute for protein-rich food in humans and as well as animal feeds.
The term single cell protein was first coined by a group of scientists at Massachusetts
Institute of Technology (MIT) during 1996.

On an average microbial biomass contains around 45 to 55% of protein, although in some

types or strains of bacteria protein content is as high as 80%. The microbial biomass also
contains other types of essential nutrients required in the human diet as well as animal feed.

History:
In the 1960s, researchers at British Petroleum developed what they called "proteins-from-oil
process": a technology for producing single-cell protein by yeast fed by waxy n-paraffins, a
product produced by oil refineries. Initial research work was done by Alfred Champagnat at
BP's Lavera Oil Refinery in France; a small pilot plant there started operations in March in
1963, and the same construction of the second pilot plant, at Grangemouth Oil Refinery in
Britain, was authorized. The term SCP was coined in 1966 by Carroll L. Wilson of MIT. The
"food from oil" idea became quite popular by the 1970s, with Champagnat being awarded the
UNESCO Science Prize in 1976, and paraffin-fed yeast facilities being built in a number of
countries. The primary use of the product was as poultry and cattle feed. The Soviets were
particularly enthusiastic, opening large "BVK" (belkovo-vitaminny kontsentrat, i.e., "protein-
vitamin concentrate") plants next to their oil refineries in Kstovo (1973) and Kirishi (1974).
The Soviet Ministry of Microbiological Industry had eight plants of this kind by 1989, when,

Hiren K. Patel 2014

 Single Cell Protein 3

pressured by the environmentalist movements, the government decided to close them down,
or convert to some other microbiological processes.

Advantages of using microorganisms

 MO grow at very fast rate under optimal conditions
 Quality and quantity is better than higher plants and animals
 Wide range of raw materials can be used
 Culture and fermentation conditions are simple
 MO can be genetically manipulated
 Microbes as Single Cell Protein Source
List of microbes used for SCP
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
Yes (U.K.), Symba
Candida utilis (TorulaYeast) 1. Confectionery -
process
2. Ethanol Feed Yes (USA)
Yes (Europe, USA,
3. Sulphite liquor -
Russia)
C. intermedia Whey - Yes; Vienna process
C. krusei (+
Whey - Yes; Kiel process
Lactobacillusbulgarius)
n-alkanes (C10 -
C. lipolytica - Yes (Russia)
C23) + ammonia
Yes (France); Le Bel
Kluyveromyces fragilis Whey Food
process
Saccharomyces cerevisiae Molasses (Food)* Yes
Fungi
Chaetomium cellulolyticum Cellulosic wastes - Promising
Fusarium graminearum Starch hydrolysate Yes (U.K)
Yes
Paecilomyces varioti Sulphite liquor -
(Finland); Pekiloprocess
Bacteria
C1 - C4
Brevibacterium sp. - Process developed
hydrocarbons
Methylophilusmethylotrophus Methanol Feed Yes (U.K.),

Hiren K. Patel 2014

 Single Cell Protein 4

Average composition of the main groups of micro-organisms (%

dry weight)

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 Acids 7-10 3-8 6-12 8-12

SCP from different substrate

SCP from N. Alkanes

In the late 1950's, British Petroleum (BP) became interested in the growth of a micro-
organism in C12-C20 alkanes. This constitutes the wax fraction of gas oils for treating. Some
crude oils contain up to 15% in wax, and these waxes must be removed since they make oil
more viscous, precipitate out at low temperatures, block tubes etc. BP uses two yeasts,
Candidor lipolytica and C. tropicals and built a 16,000 tons/year plant in Cap Lavera, France,
and a 4,000 tons/year plant in England. The product produced was called "TOPRINA". In the
UK the product "TOPRINA G" was a purer product while the one in France was not
separated from alkanes.

Both processes employed NH3 as N-source and Mg ions to increase yields. No other carbon
source was used. For 12 years TOPRINA was tested for toxicity and carcinogenecity and was
marketed as a replacement for fish meal in high protein feeds and as a replacement for
skimmed milk powder in milk replacers. There were no signs at all for toxicity or
carcinogenicity. In spite of this, people were concerned that aromatic hydrocarbons may be
carried over to SCP. The main opposition came from Japan, where environmental groups and
university professors condemned SCP as dangerous, and the matter became political. In 1972
a specialised committee decided that SCP was only for animal feeding but later, Japan was
the first country to ban petrochemical protein. Meanwhile BP and an Italian company
constructed a 100,000 tn/year plant in Sardinia.

Japanese attack on SCP, there has been great concern about and opposition to the use of SCP
from environmental groups in government. The Italian government ordered further studies
which showed that there was no hazard or carcinogenesis due to SCP. Pigs fed on 30%

Hiren K. Patel 2014

 Single Cell Protein 5

TOPRINA in their diets showed less n-paraffins in their fat tissue than those fed on pasture.
Based on this evidence the Italian government agreed to the use of TOPRINA in limited
amounts and only for export. In 1977 Italy stopped the SCP production for alkanes altogether
due to the increase in oil prices. The price of soya was more competitive. Now there is no
factory which produces any petrochemical protein.

SCP from Methane

Methane is cheap, abundant and without the toxicity problems of alkanes. It is a constituent
of North Sea Gas and is also produced during anaerobic digestion. Methane contains the most
highly reduced form of carbon and consequently gives high cell yields relative to the amount
of gas consumed. The general Methylomonas and Methylococcus have been recognised as
utilising methane as a carbon source. The species which has been extensively studied is
Methylomonas methanica. Nitrates or ammonium salts can serve as N-source. Perhaps the
most important work in this field was carried out by Shell in England. The process involves
methane oxidation by stable mixed cultures. These were

1. a methane utilising G(-) rod;

2. a Hyphomicrobium;
3. two g(-) rods; Acinetobacter and Flavobacterium

This mixed culture was one of the best examples of symbiosis. The process began in 1970 in
a 300 e pilot plant at Sittingbourne, UK. In 1974, Shell announced plans for a construction of
a larger pilot-plant in the same area and a development program in Amsterdam with a goal of
producing 100,000 tn/year. In spring 1976, Shell stopped commercialisation and its
development plans were indefinitely postponed. This decision was based on 3 factors:

1. the low price of soybeans & maize;

2. the potential of many countries for expanding existing protein sources;
3. the difficulty in applying Shell's sophisticated process in underdeveloped countries.

SCP from Methanol

The technology of SCP from methanol has been well studied and the most advanced process
belongs to ICI. The fermentation was carried out in a big airlift fermentor with the bacterium.
Methylophilus methylotropha. This organism was selected among other methanol utilises
after screening tests for pathogenicity and toxicity. As a nitrogen source ammonia was used.
The product was named "PRUTEEN". Pruteen contained 72% crude protein and was
marketed for feed as a source of energy, vitamins and minerals as well as a highly balanced
protein source. The methionine and lysine content of Pruteen compared very favourably with
white fish meal. ICI has commissioned a 60,000 tn/year plant utilising the single largest
fermentor in the world (2 x 10,000,000 l).

Unfortunately Pruteen now cannot compete with soya and fish meal. ICI hopes to be able to
sell their technology, because they have given up the idea of making money out of Pruteen.
So today Pruteen although a major engineering success is not economical to run.

Hiren K. Patel 2014

 Single Cell Protein 6

SCP from Ethanol

Ethanol although expensive as a substrate has been used for SCP. The process comes from
the Amoco Company in the US utilising a food grade yeast: "Torula". The product is sold by
the name "TORUTEIN" and government clearances have been obtained to market Torutein in
Canada and Sweden. The yeast is about 52% protein and due to its relatively low Methionine
level has a PER of about 1.7. The PER of wheat from 1.1 to 2.0. Torutein is being marketed
as a flavour enhancer of high nutritional value, and a replacement for meat, milk and egg
protein.

However it is not very successful in the United States since soya which is plentiful and cheap
can serve as an alternative or substitute to meat and egg diets.

Mycoprotein

This is a development of Ranks Hovis McDougall and is the only mycoprotein (except edible
mushrooms) that has been cleared for human consumption. It uses a Fusarium graminearum
growing in molasse, or glucose. The medium contains NH3 for nitrogen source and pH
control. The product is heat treated for RNA reduction. The mycelium is separated by
vacuum filtration, and can be technologically treated to match food texture. In the UK it is
marketed as pies and is considered a success since having less fat than meat, it can be sold at
a premium price.

SCP from Lignocellulose

The lignocellulosic wastes, mainly from agriculture, constitute the most abundant substrate
for SCP which is also renewable. The world annual production of straw for example reaches
600 million tons every year. In Greece the straw from wheat and rye, the two most important
cereals, is an estimated 1.5 million tons per year. For the utilisation of lignocellulose, a pre-
treatment is usually necessary. Many pre-treatment methods have been reported which vary
from alkali or acid treatment, steam explotion or even x-ray radiation.

To the present time the only economical utilisation of lignocellulosic wastes is in mushroom
production. Besides our well know cultivated mushroom Agaricus bisporus there are many
important ones which contain lignocellulolytic enzymes and are cultivated for food mainly in
Asia and Africa. Some are of great economic significance and are cultivated on an industrial
scale. Examples of important ones include the following species: Volvariella sp., Lentinus
edodes and Pleurotus sp.

Single Cell Protein Production Process

Single cell Protein Production process follows these steps

1. Microbial Screening
2. Choice of Raw Materials
3. Process Engineering and Process Optimization
4. Technology Development

Hiren K. Patel 2014

 Single Cell Protein 7

5. Economic Consideration / Process Feasibility

6. Safety Concerns

1. Microbial screening:

Microbial Screening is the first step in Production process, suitable microbe which yields
good amount of protein need to be selected. microbial strains are collected from various
habitats like soil, water, air and or from other biological materials. Microbes are selected by
various studies including mutagenisis and other genetic methods, some times wild types are
also used.

2. Choice of Raw Material:

This part is little cumbersome and one need to focus on the correct composition of carbon
suppliment which yields higher biomass production in lesser time need to be analyzed.
various carbon sources are like wood waste, straw, other food processing wastes are also can
be tried to optimize higher biomass production. Substrates for Single Cell Protien Production
can be subdivided into three categories:

 High energy sources (natural gas, n-alkanes, gas-oil, methanol, ethanol, acetic acid);
 Various wastes (molasses, sulfite waste liquor, milk, whey, fruit wastes); and
 Renewable plant resources (sugar, starch, cellulose).

3. Process engineering:

The technical conditions of cultivation for the optimized strains are done and all metabolic
pathways and cell structures will be determined.

4. Technology Development:

Technology development is the next step where the adoption of the technical performance of
the process in order to make the production ready for use on the large technical scale.

5. Economic factors:

Energy consumption, cost of production are the important factor while going for large scale
production phase, this need to be thoroughly analyzed and an energy efficient process need to
be developed or else it will end up with loss.

6. Safety demands & Environmental protection:

Since the SCP produced is for human consumption or for feeding animals safety of the
product need to be tested. certain microbes produces toxic compounds which can have
determinal effect on humans and also for the environment, so the whole process should be
monitored properly.

The Problem of Nucleic Acids

Hiren K. Patel 2014

 Single Cell Protein 8

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 (78-25 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. 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:

1. chemical treatment with NaOH;

2. treatment of cells with 10% NaCl;
3. thermal shock.

These methods aim to reduce the RNA content from about 7% to 1% which is considered
within acceptable levels.

Advantages of Single Cell Protein

Advantages of using microbes for Large scale production of Single cell proteins are

1. Single cell protein high protein and low fat content.

2. Single cell protiens are good source of vitamin.
3. It can be produced through out the year.
4. Generation time of microbes are less, ie, they multiply rapidly building up the
biomass, more the biomass more the protein source.
5. Protein content is very high in dried biomass upto 85%
6. During the production of SCP biomass, certain microbes produce usseful byproducts
such as organic acids.
7. Waste (wood waste, food processing waste, hydrocarbons, etc) can be used as a
source for carbon for growing Microbes there by having advantage of environmental
clean up also.
8. Doesn't require sophisticated lab setup for algae and certain other microbes.
9. High efficiency substrate conversion.

Disadvantages of Single Cell Protein

Even though it single cell proteins have the above mentioned advantages, it has some
disadvantages also, the major problem associated with the use of single cell proteins are

1. Many microbes produce various toxic compounds, so consumption of such toxic can
have serious effect on health of humans (Food Grade SCP), or in animals (Feed).
2. Single Cell Protein diet suppliments can pose allergic reaction.

Hiren K. Patel 2014

 Single Cell Protein 9

3. Consuming SCP, in-taking higher amount of Nucleic acids which can lead to
gastrointestinal problems.
4. Food grade SCP production are expensive due to the need to maintain high level
sterility conditions in the production facility.

Sources of SCP and its disadvantages

1. Algae:
 Chlorella, Scenedesmus acutus and Sprulina 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 Thaiwan,
Thailand, Japan, Israel, Mexico and USA.
 It is spray dried and sold as pills and powders.
 Spirulina grown on sewage water is free of pathogenic microorganisms.
Disadvantages:
 These are not suitable for human consumption because they are rich in Chlorophyll.
(Except Spirulina)
 It has low density i.e. 1-2 gm dry weight/litre of substrate.
 There is lot of risk of contamination during growth.
2. Yeasts and Fungi
 The filamentous fungi such as Chaetomium celluloliticum – grows on cellulose waste,
Fusarium graminearum – grows on starch and Paecilomyces varioti – grows on
sulphur liquar are used for the production of SCP. These have about 50 – 55 %
protein.
 Yeasts such as Candida utilis (Torula yeast), Candida lipolytica – grow on Eathanol
and Saccharomyces cervicea – grows on Molasses are used for SCP production.
 Torula yeast as a food is obtained through fermentation using molasses as substrate. 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.
 Several species of Mushrooms are used as protein rich food
Disadvantages:
 These have high nucleic acid content.
 Filamentous fungi show slow growth rate than yeasts and bacteria.
 There is contamination risk.
 Some strains produce mycotoxins and hence they should be screened.
3. Bacteria
 These have more than 80% protein. They are poor in sulphur containing amino acids.
 Brevibacterium uses hydrocarbons as substratum and Methylophilus methylitropous
uses methanol.
 It has high nucleic acid content
Disadvantages:
 It has high RNA content.
 Risk of contamination is very high during the production process.
 Recovering the cells is a bit problematic.
 Endotoxin production should be carefully tested.

Hiren K. Patel 2014

 Single Cell Protein 10

Problems with SCP

 Despite the very attractive features of SCP as a nutrient for humans there are many
problems that deter its adoption on a global basis. Such problems include high
concentration of nucleic acids (6-10) % which elevates serum uric acid levels and
results in kidney stone formation.
 About 70 to 80% of the total Nitrogen is represented by amino acids while the rest
occur in nucleic acids. This concentration of nucleic acid is higher than other
conventional protein and it is characteristics of all fast growing organisms.
 The problem which occurs from the consumption of protein with high nucleic acid
concentration (18-25g/100g protein dry weight) is the production of high
concentration of uric acid in the blood causing health disorders such as gout and
kidney stone.
 It has also been noted that the cell wall of the microorganisms may be non digestible,
there may be unacceptable color and flavors (especially in algae and yeast), their cells
should be killed before consumption, there may also be possible skin reactions from
consumption of foreign protein and gastrointestinal reactions may occur resulting in
nausea and vomiting.
 SCP from algae may not be suitable for human consumption because they are rich in
chlorophyll, (except Spirulina), also it has low density i.e. 1-2 gm dry weight/litre of
substrate and there is lot of risk of contamination during growth.
 SCP from yeast and fungi has high nucleic acid content, the filamentous fungi show
slow growth rate than yeasts and bacteria there is high contamination risk and some
strains produce mycotoxins and hence they should be well screened before
consumption. All these detrimental factors affect the acceptability of SCP as global
food.

Hiren K. Patel 2014