Single Cell Protein: Production and Process
Single-cell proteins are the dried cells of microorganism, which are used as protein supplement in human foods or animal feeds. Microorganisms like algae, fungi, yeast and bacteria, utilize inexpensive feedstock and wastes as sources of carbon and energy for growth to produce biomass, protein concentrate or amino acids. Since protein accounts for the quantitatively important part of the microbial cells, these microorganisms, also called single cell protein as natural protein concentrate. With increase in population and worldwide protein shortage the use of microbial biomass as food and feed is more highlighted. Although single cell protein has high nutritive value due to higher protein, vitamin, essential amino acids and lipid content, there is a doubt to be replaced to the conventional protein sources due to their high nucleic acid content and slower in digestibility. They also may be considered as foreign material by body, which may subsequently results in allergic reactions.
April 12, 2010; Accepted: April 28, 2010;
Published: July 06, 2010
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,
||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
Yeast was the first microorganism whose importance as animal feed supplement
was recognized almost a century ago. During World War I, Germany replaced half
of imported protein sources by yeast. Pruteen was the first commercial single
cell protein used as animal feed additive. From a nutritional viewpoint, Nucleic
Acids (NA) content in SCP is one of the main factors hindering its utilization
as food. Excessive intakes of NA lead to uric acid precipitation, causing health
disorders, such as gout or kidney stone formation.
So, in human their NA contents must be reduced below 2%. Several technologies
have been reported to reduce the NA content of microbial cells, including both
chemical and enzymatic procedures. Chemical and enzymatic methods have disadvantages
(Parajo et al., 1995). Various microorganisms
used for the production of SCP are bacteria (Cellulomonas, Alcaligenes,
etc.), algae (Spirulina, Chlorella, etc.), molds (Trichoderma,
Fusarium, Rhizopus, etc.) and yeast (Candida, Saccharomyces,
etc.). Microorganisms can utilize a variety of substrate like agricultural wastes
and effluents, industrial wastes, natural gas like methane, etc. that also help
in decomposing pollutants (Huang and Kinsella, 1986).
SINGLE CELL PROTEIN DEFINITION
A variety of microorganisms and substrate are used to produce single cell proteins.
Yeast is suitable for single cell protein production because of its superior
nutritional quality (Table 1). The supplementation cereals
with single cell proteins, especially yeast, make them as good as animal proteins
(Huang and Kinsella, 1986). The necessary factor considered
for use of SCP is the demonstration of the absence of toxic and carcinogenic
compounds originated from the substrates, biosynthesized by the microorganisms
or formed during processing. High nucleic acid content and low cell wall digestibility
are two of the most important factors limiting nutritional and toxicological
value of yeast for animal or human consumption (Alvarez
and Enriquez, 1988). As constituents of nucleic acid, purine compounds in
human diet mostly metabolized to yield uric acid whose high concentration may
lead to gout or renal stones. However, nucleic acid is not a toxic component
and it causes only physiological effects at higher levels like any other essential
dietary ingredients taken in larger amounts. It has been calculated that 100
lbs of yeast will produce 250 tons of proteins in 24 h. Algae grown in ponds
can produce 20 tons (dry weight) of protein, per acre, per year. Bacteria are
usually high in protein (50 to 80%) and have a rapid growth rate. The principal
disadvantages are as follows:
||Bacterial cells have small size and low density, which makes
harvesting from the fermented medium difficult and costly
||Bacterial cells have high nucleic acid content relative to
yeast and fungi. To decrease the nucleic acid level additional processing
step has to be introduced and this increases the cost
||The general public thinking is that all bacteria are harmful
and produce disease. An extensive education program is required to remove
this misconception and to make the public accept bacterial protein
Yeasts have advantages such as their larger size (easier to harvest), lower
nucleic add content, high lysine content and ability to grow at acidic pH. However,
the most important advantage is familiarity and acceptability because of the
long history of its use in traditional fermentations. Disadvantages include
lower growth rates, lower protein content (45 to 65%) and lower methionine content
than in bacteria. Filamentous fungi have advantages in ease of harvesting, but
have their limitations in lower growth rates, lower protein content and acceptability.
Algae have disadvantages of having cellulosic cell walls which are not digested
by human beings. Secondly, they also concentrate heavy metals. In the case of
algae it has to be stressed that, due to technical and economical reasons, it
is not the general intention to isolate and utilize the sole protein, but to
propagate the whole algal biomass. So, the term SCP is not quite correct, because
the micro-algal material is definitely more than just protein. To date, world-wide
various sophisticated technologies are employed for mass production and processing
of photoautotrophic microalgae. The annual world production of all microalgae
species is estimated to about 10,000 tons year-1 (Becker,
1994, 2007; Richmond, 2004).
The algal biomass as sun dried or in compressed form as pastilles is the predominant
product in microalgal biotechnology (Radmer, 1996).
More than 75% of the annual microalgal biomass production is used for the manufacture
of powders, tablets, capsules, or pastilles. This biomass is harvested from
natural waters or artificial ponds or photobioreactors (PBR) and subsequently
separated from the growth media followed by drying (Renaud
et al., 1994). The two major species cultivated for this purpose
are the unicellular green alga, Chlorella and more recently, filamentous
blue-green alga (Cyanobacterium), Spirulina (Raja
et al., 2008). The production of SCP from various microbes, particularly
from fungi and bacteria has received considerable attention, in contrast, only
a few studies have dealt with the feasibility of using SCP from microalgae (Mahasneh,
1997). Algal proteins are of high quality and comparable to conventional
vegetable proteins. However, due to high production costs as well as technical
difficulties, cultivation of algae as protein is still in evaluation (Rasoul-Amini
et al., 2009). The celluloid cell wall, which represents about 10%
of the algal dry matter, poses a serious problem in digesting/utilizing the
algal biomass, since it is not digestible for humans and other non-ruminants.
Hence, effective treatments are necessary to disrupt the cell wall to make the
protein and other constituents accessible for digestive enzymes. Several authors
have studied the effect of different post-harvesting treatments on the digestibility
of various algal species (Becker, 2007). Different species
of algae, fungi, yeasts and bacteria are used as single cell protein and produced
at commercial scale (Table 2). These organisms are grown on
different carbon sources. Although, microalgae with some other supplements have
been used as an essential food for the larval stages of fish and shellfish.
Yeast cells have been considered as a substitute because of their small particle
size, high protein content as SCP and relatively low production costs (Kim
et al., 1998). However, poor digestibility may be an important constraint
in the use of this SCP as a food source in seed production of aquacultural organisms,
since yeast has a complex and thick cell envelope. The external mannoprotein
layer of the yeast cell wall is probably the major barrier to digestion (Rumsey
et al., 2007; Kim and Chung, 2001). Several
methods have been developed to improve the digestibility of SCP products: mechanical
disruption, autolysis and enzymatic treatment (Curran et
al., 1990). For maintenance of a stock collection of microorganisms
preservation by drying or freeze-drying for long periods of time was reported
to be good for yeast or algae (Cordero and Voltolina, 1997;
Kim and Chung, 2001). It has been agreed that the criteria
used to evaluate SCP production are growth yield, total protein (39-73%) and
nucleic acid contents (1-11%). Marine yeasts are better candidates for marifeed
production according to their easy cultivation in the fermenter, high cell density
and high content of essential amino acids. The substrates which have been used
for SCP production by yeasts so far include sorghum hydrolysate, sulfate waste
liquor, pawn-shell wastes, dairy wastes, methanol, molasses, starch and plant
origin liquid waste. Several fungi like Fusarium oxyporum var., lini
and Chetomium cellulolyticum, algae like Chlorella and Spirulina,
yeast like Candida lipolytica and Saccharomyces sereviciae and
phototrophic bacteria like Rodospirillum sp., had been explored for SCP.
It is also said that Scytalidium acidophilum compares favorably with
the values for the well-known single-cell protein producer Candida utilis.
Kluyveromyces marxianus produces enzyme that can be used to modify whey
for the production of single cell protein (Chandrani-Wijeyaratne
and Tayathilake, 2000). Spirulina maxima is grown on a commercial scale
in Lake Texcoco in Mexico. The quality of SCP is an important factor for commercial
production. 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. Estimation
of the Biological Value (BV) is a measure of nitrogen retained for growth or
maintenance. Another parameter, which reflects the quality of a protein, is
the Digestibility Coefficient (DC). 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.
The SCP generally values in terms of Kjeldhal nitrogenx6.25 (standard factor
relating amino nitrogen to protein content). However, about 10-15% of the total
nitrogen in fungi and yeasts is in the form of nucleic acids (Riviere,
The worldwide, large-scale development of SCP processes has contributed greatly
to the advancement of present day biotechnology. Research and development of
SCP processes has involved work in the fields of microbiology, biochemistry,
genetics, chemical and process engineering, food technology, agriculture, animal
nutrition, ecology, toxicology, medicine and veterinary science and economics.
In developing SCP processes new technical solutions for other related technologies
in waste water treatment, production of alcohol, enzyme technology and nutritional
science also improves. The future of SCP will be heavily dependent on reducing
production costs and improving quality by fermentation, downstream processing
and improvement in the producer organisms as a result of conventional applied
genetics together with recombinant DNA technologies (Omar
and Sabry, 1991). Single cell proteins have application in animal nutrition
as: fattening calves, poultry, pigs and fish breading in the foodstuffs area
as: aroma carriers, vitamin carrier, emulsifying aids and to improve the nutritive
value of baked products, in soups, in ready-to-serve meals, in diet recipes
and in the technical field as: paper processing, leather processing and as foam
stabilizers. The production of single cell protein takes place in a fermentation
process. This is done by selected strains of microorganisms which are multiplied
on suitable raw materials in technical cultivation process directed to the growth
of the culture and the cell mass followed by separation processes. Process development
begins with microbial screening, in which suitable production strains are obtained
from samples of soil, water, air or from swabs of inorganic or biological materials
and are subsequently optimized by selection, mutation, or other genetic methods.
Then the technical conditions of cultivation for the optimized strains are done
and all metabolic pathways and cell structures will be determined. Besides,
process engineering and apparatus technology adapt the technical performance
of the process in order to make the production ready for use on the large technical
scale. Here is where the economic factors (energy, cost) come into play. Safety
demands and environmental protection is also considered in the production of
SCP in relation both to the process and to the product. Finally, safety and
the protection of innovation throw up legal and controlled aspects, namely operating
licenses, product authorizations for particular applications and the legal protection
of new process and strains of microorganisms (Steinkraus,
The classical raw materials are substances containing mono and disaccharides,
since almost all microorgansims can digest glucose, other hexose and pentose
sugars and disaccharides. These materials also are utilized in other branches
of industry with a high price level, which puts the economic aspect of the production
of microbial biomass in doubt (Oura, 1983). The choice
of substrates that are normally abundant has determined the design and strategy
of SCP processes. The most widespread and commonly used substrates for SCP production
have been those where the carbon and energy source is derived from. Many companies
producing SCP including BP (UK), Kanegafuichi (Japan) and Liquichimica (Italy)
appeared on the scene. In the United States less than 15% of the plants making
SCP were said to rely on hydrocarbons as the source of carbons and energy for
the micro-organisms. Other potential substrates for SCP include bagasse, citrus
wastes, sulphite waste liquor, molasses, animal manure, whey, starch, sewage,
SCP derived from high energy sources: Materials with high commercial
value as energy sources or derivatives like gas oil, methane, methanol and n-alkanes
are of interest in SCP production. The microbes involved are mostly bacteria
and yeast and several processes are now in operation. The wisdom of using such
high-energy potential compounds for food production has been questioned by many
scientists. British Petroleum uses two yeasts, Candida lipolytica and
C. tropicalis and C12-C20 alkanes as substrate
which is of the wax fraction of gas oils for treating. Some crude oils contain
up to 15% in wax. The product produced was called TOPRINA. 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. After a while the main opposition came from Japan. In 1972
a specialised committee decided that SCP was only for animal feeding but later,
Japan was the first country to ban petrochemical protein. In 1977 Italy stopped
the SCP production from alkanes due to the increase in oil prices. The price
of soya was more competitive. Now there is no factory which produces any petrochemical
protein. Methane as a SCP source has been extensively researched. The species
which has been extensively studied is Methylomonas methanica and nitrates
or ammonium salts can serve as N-source. The technology used for this purpose
faced too many technical difficulties to optimize the production. In contrast,
methanol offers great economic SCP interest. A large-scale fermentation plant
for producing the methanol-utilizing bacterium Methylophilus methtlotrophus
was constructed by ICI (Imperial Chemical Industries Company), UK. The ICI SCP
protein was used exclusively for animal feeding. Methanol as a carbon source
for SCP has many inherent advantages over n-paraffins, methane gas and even
carbohydrates composition is independent of seasonal fluctuations. There are
no possible sources of toxicity in methanol and it dissolves easily in the aqueous
phase in all concentrations. The ICI Pruteen (72% protein) plant (by Pseudomonas
methylotrophus) was the only process of its kind in the Western world but
could not operate economically at present methanol prices and cannot compete
with soya and fish meal so has ceased production. Methanol represents approximately
50% of the costs of the product. In the USA the cost of SCP derived from methanol
is two-to five-folds the cost of fishmeal. In the Middle East the low cost of
methanol and higher costs of fishmeal coupled with a need to produce more animal
products could make SCP an attractive proposition. Ethanol is a particular suitable
source if the SCP is intended for human consumption. The process comes from
the Amoco Company in the US utilizing food grade yeast: Torula. The product
is sold by the name "Torutein" even in Canada and Sweden. The yeast is about
52% protein. Torutein is being marketed as a flavor 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. In the foreseeable
future the comparative status of ethanol SCP will depend on local factors: over-capacity
in ethylene crackers, agricultural carbohydrates surpluses and political decisions
about regional economic independence and foreign trade balances (Trinci,
1992). The use of n-alkanes as substrate for SCP has been extensively studied
many countries and represents a very complex biotechnological process. However,
most of these processes have now ceased operation because of suspected health
hazards resulting from the presence of carcinogens in the SCP. The massive technology
developed in this field in Japan and other Eastern countries has been turned
over to the study of alcohol- based SCP and SCP form organic wastes. Many genera
of yeasts and molds utilize aliphatic hydrocarbons (alkanes and alkenes) for
growth and for SCP production. These microorganisms secrete emulsifying substances
during fermentation which increase the solubility of alkanes and alkenes. These
substrates are transported by passive diffusion into the cell. The higher lipid
content in these microorganisms facilitates the hydrocarbon transport. For large-scale
production of SCP, gas oil and C10-C17 alkanes were used
as substrate. Gas oil fermentation has been carried out in airlift fermenter
using Candida tropicalis and C. lipolytica (Riviere,
SCP from wastes: The amount of agricultural and some industrial wastes
used for SCP production can be locally very high and may contribute to a significant
level of pollution in watercourses. Thus, the utilization of such materials
in SCP processes serves two functions as reduction in pollution and creation
of edible protein. Cellulose from agriculture and forestry sources constitutes
the most abundant renewable resource in the planet as potential substrates for
SCP production. Cellulose has emerged as an attractive substrate for SCP production
but in nature it is usually found with lignin, hemicellulose, starch, etc.,
in a complex form. Therefore, if cellulose is to be used as substrate it must
be pretreated chemically (acid hydrolysis) or enzymatically (cellulases) to
remove cellulose as fermentable sugars (Callihan and Clemmer,
1979). For the utilization of lignocellulose, a pre-treatment is usually
necessary. Many pre-treatment methods have been reported which vary from alkali
or acid treatment, steam explosion or even x-ray radiation. To the present time
the only economical utilization of lignocellulosic wastes is in mushroom production.
Besides our well known 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
Volvariella sp., Lentinus edodes and Pleurotus sp. In the
manufacture of industrial cellulose for paper and tissue production the cost
of these steps has prevented the generalized production of SCP from cellulose
(Callihan and Clemmer, 1979). Wood can be also cooked
in a medium containing calcium sulfite with excess free sulfur dioxide. Lignin
is thus converted to lignosulfonates and hemicellulose is hydrolysed to monosaccharides
and may be further broken down to furfurols. The amount of free sugars in the
spent liquor is variable with the type of procedure chosen, as various cellulose
fibers may be obtained with different degrees of degradation. Spent sulfite
liquor has been used as a substrate for fermentations since 1909 in Sweden and
later in many other parts of the world. The first organism to be used was Saccharomyces
cerevisiae, although this organism is unable to metabolise pentoses which
are found in considerable amounts in this waste product. Later, other organisms
better suited for the assimilation of all the sugar monomers were chosen, namely
Candida tropicalis and Candida utilis. Yeast produced from sulfite
liquor has been used for feeding at war periods, but lost favour in peace time.
However, experiences of bakers yeast produced from sulfite liquor exist in
Finland by Peliko process. The protein content of the fungus Paecilomyces
variotii exceeds 55% (w/w) and has been officially approved as a food in
Finland. In 1983, the projected biomass production of the process was estimated
to be around 7000 tons per year (Oura, 1983). Nowadays,
extracellular cellulases are commercially used in cellulose separating process.
Cellulase is a complex of three enzymes (endocellulase, cellobiohydrolase, cellobiase).
The cost of production of these extracellulases is about $ 0.011 L-1
of crude enzyme and $ 0.11 L-1 of purified enzyme. The cost of extraction
of fermentable sugar using cellulase is about $ 0.50 kg-1 sugar.
A number of efficient cellulase producers have been reported but Trichoderma
viride continued to be well known high cellulase-producing organism. Chaetomium
cellulolyticum is another cellulolytic fungus which grows faster and forms
80% more biomass-protein than Trichoderma. This means that C. cellulolyticum
is suitable for SCP production while T. viride is a hyper producer
of extracellular cellulases. The amino acid composition of C. cellulolyticum
is generally better than that of T. viride and similar to alfalfa
and soya meal protein (Bhalla et al., 2007). A
cheaper, more amenable SCP substrate of carbohydrate origin is starch. This
very abundant carbohydrate may be obtained from rice, maize and cereals. In
tropical countries, cassava has been proposed as a good source of starch for
SCP processes. The Symba process developed in Sweden utilized starchy wastes
combining two yeasts in sequential mixed culture: the amylase producing Endomycopsis
fibuligira and the fast growing Candida utilis. The process consists
of three phases: The incoming starch waste is fed through heat exchangers and
sterilized. The medium is then fed to a first bioreactor where the starch hydrolyzing
yeast grows and hydrolyses starch. The hydrolyzed solution is then fed to a
second reactor where culture conditions favor the proliferation of C. utilis.
Whey traditionally originates from the curding process in cheese production,
but can now be obtained after ultra filtration procedures for the production
of spreading cheeses, where the protein fraction corresponding to lactalbumins
and lactoglobulins is incorporated to the casein fraction and all the proteins
are in native form with the principal component as lactose (4-6%) (w/v). Although
other nutrients can be find in significant amounts (Moebus
and Teuber, 1983). Whey has been presented as an extremely suitable substrate
for the production of SCP. In 1956 The French dairy company Fromageries Bel
pioneered a project to produce yeast from whey, using lactose assimilating Kluyveromyces
marxianus (formerly K. fragilis) (Oura, 1983;
Moulin et al., 1983). Molasses is a by-product
of the sugar manufacturing process. Biomass production from molasses requires
supplementation with a suitable nitrogen source, as well as phosphorus (Callihan
and Clemmer, 1979). The Ranks Hovis McDougall mycoprotein (except edible
mushrooms) 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. Coffee-pressing wastes contain soluble carbohydrates
and have a high chemical oxygen demand (COD) and soluble solid contents. In
Guatemala, Trichoderma sp. is used to produce SCP on this substrate.
Cellulomonas grows on bagasse and Thermoactinomyces on fermented
livestock wastes (Humphrey, 1975).
Fermentation process: The fermentation process requires a pure culture
of the chosen organism that is in the correct physiological state, sterilization
of the growth medium which is used for the organism, a production fermenter
which is the equipment used for drawing the culture medium in the steady state,
cell separation, collection of cell free supernatant, product purification and
effluent treatment. A fermenter is the instrument, which is set up to carry
out the process of fermentation mainly the mass culture of plant or animal cells.
Fermenters can vary in size from laboratory experimental models of one or two
litres capacity, to industrial models of several hundred litres capacity. A
bioreactor is different from a fermenter as it used for the mass culture of
microorganisms. The chemical compounds synthesised by these cultured cells such
as therapeutic agents can be extracted easily from the cell biomass. The design
engineering and operational parameters of both fermenters and bioreactors are
identical. Fermenters and bioreactors are also equipped with an aerator, which
supplies oxygen to aerobic processes also a stirrer is used to keep the concentration
of the medium the same. A thermostat is used to regulate temperature and a pH
detector and some other control devices, which keep all the different parameters
needed for growth constant (Ferrianti and Fiechter, 1983;
Sinclair and Cantero, 1990). For the producing and harvesting
of microbial proteins cost is a major problem. Such a production even in high
rate causes dilute solutions usually less than 5% solids. 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. The removal of the amount of water that is necessary to make
the material stable for mass storage is not economically viable. Single cell
proteins need to be dried to 10% moisture or they can be condensed and denatured
to prevent spoilage (Sinclair and Cantero, 1990). The
physiological features of microbial organisms recommend the control of the carbon
source concentrations, as a limiting substrate, as well as an adequate supply
of oxygen for the maintenance of balanced growth under an oxidative metabolic
pattern. However, since microbial growth is a time dependent process, it exerts
continuous modifications on all process parameters which influence physiology,
but most dramatically, over substrate concentration. Therefore, an adequate
technology which maintains appropriate growth conditions for a prolonged period
of time must be implemented specifically for the purpose of obtaining high yield
and productivity values. Batch fermentations are clearly inadequate for the
purpose of biomass production, since the conditions in the reaction medium change
with time (Oura, 1983). Fed-batch fermentations are better
suited for the purpose of biomass production, since they involve the control
of the carbon source supply through feeding rates. However, as the biomass concentration
increases, the oxygen demand of the culture reaches a level which cannot be
met in engineering or economic terms. Fed-batch culture is still in use for
bakers yeast production using well established and proven models (Steinkraus,
1986). However, they have not been favored for the production of SCP at
a large industrial scale. Prolonging a microbial culture by continuous addition
of fresh medium with the simultaneous harvesting of product has been implemented
successfully in industrial fermentations destined to biomass production. The
most commonly used principle has been the chemostat: a perfectly mixed suspension
of biomass into which medium is fed at a constant rate and the culture is harvested
at the same rate so that the culture volume remains constant. The technical
implications of chemostat culture are various and extremely relevant (Cooney,
1986). Production periods as long as six weeks have been implemented in
many fungal and yeast (Forage and Righelato, 1979). A
common problem of industrial fermentations is the profuse appearance of foam
on the head space of the reactor, causing reactor pressurization, spillages
and contamination hazard. Among the various designs which have been put to effect,
the deep-jet fermenter and the air-lift fermenter have been the most successfully
applied (Stanbury et al., 2000). Air-lift has enjoyed
the greatest success as the configuration of choice for continuous SCP production.
This is presently used in the production of myco-protein which is the basis
for Quorn products. The control of key process variables is a critical
element of SCP production, from oxygen transfer, substrate and product concentration,
to the appearance of minimal amounts of toxic compounds through undesired metabolic
processes, which may compromise the quality of the final product. The biomass
from yeast fermentation processes is harvested normally by continuous centrifugation.
Filamentous fungi are harvested by filtration (Solomons,
1985). The biomass is then treated for RNA reduction and dried in steam
drums of spray driers. Drying is expensive, but results in stabilized product
with shelf lives of years. Solid State Fermentation (SSF) is growth of microorganisms
on predominantly insoluble substrate where there is no free liquid. Generally,
under combined conditions of low water activity and presence of intractable
solid substrate, fungi show luxuriant growth. Hence, proper growth of fungi
in SSF gives much higher concentration of the biomass and higher yield when
compared to submerged fermentation. The advantage in SSF process is the unique
possibility of efficient utilization of waste as the substrate to produce commercially
viable products. The process does not need elaborate prearrangements for media
preparation. The process of SSF initially concentrated on enzyme production.
But presently, there is worldwide interest for (SCP) production due to the dwindling
conventional food resources (Zadrazil and Puniya, 1995).
For SCP production large-scale fermenters are required. So with high biomass
production, high oxygen transfer rates and high respiration rates which in turn
increase metabolic heat production and the need of an efficient cooling system
ensued. In such a continuous operation for SCP production the economics of this
production must be strongly taken into account. The Economics factors that should
be taken into account during this fermentation period are: Investment, Energy,
Operating costs, Waste, Safety and the Global market.
Substrate costs: The substrate costs are the largest single cost factor.
Simplifying the manufacture and purification of raw material can save costs.
Moreover the manufacture of raw materials is more economical in larger plants.
Factors involved in the raw materials costs are site, raw material production,
process capacity of the plant and substrate yield.
Utilities: The energy for compressing air, cooling, sterilizing and
drying forms the next most important cost factor. Sites with cheaply available
thermal, electrical, fossil or process derived energy are to be preferred.
Capital load: The capital dependent costs are determined, by the cost
of the apparatus for the process, the capacity of a plant and the capacity conditions.
The main variable here is the size of the plant. Small plants can be profitable
only if they include simplifications of processes and material to a considerable
degree. The greater expenditure on apparatus in processes with cheap, simple
and unpurified raw materials usually does not pay in comparison with more expensive
pure substrates with simpler technology. High productivities in fermentation
are compensated by the greater expenditure on energy to achieve these productivities,
so that optimum can be determined.
Product-specific variables: The process costs arising are covered only
by the product produced. The absolute value of the product is governed by the
amount of product referred to the costs involved and by the quality of the product.
The upgrading of the product may consist of purification and separation of the
ACCEPTABILITY AND TOXICOLOGY OF SCP
The name of the raw materials used in SCP processes represents the main safety
hazard. Toxicology testing of the final product must include short-term acute
toxicity testing with several different laboratory animal species, followed
by extensive and detailed long term studies. It represents a major scientific
and financial investment. The acceptability of SCP when presented as a human
food does not depend only on its safety and nutritional value. In addition to
the general reluctance of people to consume material derived from microbes,
the eating of food has many subtle psychological, sociological and religious
SCP PROCESSING FOR FOOD
The effective use of microbial protein for human food requires:
||Liberation of cell proteins by destruction of indigestible
||Reduction of nucleic acid content
Methods of cell wall destruction: The use of microorganism for refined
SCP requires not only an adequate amount of specific organism but also an efficient
means of disrupting the cell wall (Baldwin and Robinson,
1994). Mechanical integration of cell wall can be carried out either by
crushing, crumbling, grinding, pressure homogenization or ultra sonification.
Various enzymes or combination of enzymes can be used to digest and disrupt
cell wall, either partially or completely. Enzymatic hydrolysis of cell wall
is attractive in terms of its delicacy and specificity for only the cell wall
structure. It may be used as an alternative to the mechanical disruption, especially
for materials that can be inactivated during the mechanical process and it can
be performed by endogenous or exogenous enzyme from other microorganisms. However,
extensive enzymatic lysis of cells is a very slow process compared to mechanical
disruptions. It is possible to use two or more methods for cell disruptions.
Combined mechanical and enzymatic degradation of yeast cell wall was tested
by Asenjo and Dunnill (1981). In case of yeast cells
they first can be mechanically broken and then incubated with a lytic enzyme
(Damodaran and Kinsella, 1983). This resulted in the
release of a substantial amount of protein mostly from organelles and cell walls.
Baldwin and Robinson (1994) reported enhanced disruption
of Candida utilis by enzymatic pretreatment and high-pressure homogenization.
Other methods employed for yeast cell breakage include: autolysis followed by
enzymatic or alkali treatments (Benaiges et al.,
1989), NaCl induced autolysis at different temperature, chemical disintegration
using detergent such as sodium dodecyl sulfate or Triton X-100, acid or solvent
(Lee, 1996). The digestibility of yeast and microalgae
can be greatly increased by drying at high temperature under certain conditions.
However, the heat treatment needed to increase the digestibility of the cells
also affects the protein quality and other valuable cell components (Hedenskog
and Morgen, 1973). Here are the methods for cell disruption:
||Chemical treatment: acid, base, solvent, detergent
||Enzyme analysis: lytic enzymes, phage infection, autolysis
||Physical treatment: freeze-thaw, osmotic shock, heating and
||High pressure homogenization
||Pressure extrusion: french press, freeze pressing
||Decompression (pressure chamber)
Removal of nucleic acids: Several methods have been proposed to reduce
nucleic acid levels in SCP. These methods involve chemical and enzymatic treatments.
Each has disadvantages both in terms of cost and potential nutritional concern.
In 1977, the extraction of nucleic acid by acidified alcohol, salt, acid and
alkalis has been proposed. Alkaline extraction of microbial biomass at elevated
temperature was also used in 1970. The process resulted in high protein yield
with low nucleic acid. However, alkaline hydrolysis of nucleic acid at high
temperature causes the formation of potentially toxic compounds such as lysinoalanine.
It is an unusual amino acid involved in cross-linking of alkaline protein. Lysinoalanine
has been shown to reduce digestion and induce kidneys changes in rats (Damodaran
and Kinsella, 1983). It also implicated in skin allergy in some persons
consuming treated protein (Scrimshaw and Dillen, 1977).
Chemical modification of yeast nucleoproteins with anhydrides has been used
to reduce the nucleic acid levels. Yeast contains considerable amounts of endogenous
ribonulease activity that is used to hydrolyzed yeast RNA and that cause reduction
of nucleic acid level in yeast protein. At the optimum conditions of ribonuclease
activity, significant activation of endogenous protease also occurs. This result
in proteolytic degradation of protein and thereby, decreases the yield of protein.
Alternatively, nuclease has been added exogenously to reduce the nucleic acid
content of SCP. Pancreatic ribonuclease (RNase A) and a fungal ribonuclease
of Aspergillus candidus strain M16 has been used as the source of exogenous
nuclease for the reduction of nucleic acid in the cells of yeast species allowing
a substantial reduction of NA (Maul et al., 1970;
Kunhi and Rao, 1995). Bacterial or pancreatic nucleases
have been also studied for NA removal from yeast cells. Hydrolysis of
NA has also been performed by using immobilized enzymes (Parajo
et al., 1995).
Single celled protein (SCP) production, referring to the fact that most of
the micro-organisms used as producers grow as single or filamentous individuals
rather than as complex multi-cellular organism such as plants or animals. Use
of microbes in the production of proteins gives many advantages over the conventional
method methods. Microbes have shorter generation time, allow easy transformation,
utilize many substrates, have no requirements in arable land or any particular
season to grow and have the possibility of continuous production in any part
of the world. The cell yield varies according to the substrate and type of microorganism.
Alvarez, R. and A. Enriquez, 1988. Nucleic acid reduction in yeast. Applied Microbiol. Biotechnol., 29: 208-210.
CrossRef | Direct Link |
Asenjo, J.A. and P. Dunnill, 1981. The isolation of lytic enzymes from Cytophaga sp. and their application to the rupture of yeast cells. Biotechnol. Bioeng., 23: 1045-1056.
Baldwin, C.V. and C.W. Robinson, 1994. Enhanced disruption of Candida utilis using enzimatic pretreatment and high pressure homogenization. Biotechnol. Bioeng., 43: 46-56.
PubMed | Direct Link |
Becker, E.W., 1994. Microalgae: Biotechnology and Microbiology. Cambridge University Press, Cambridge, UK.
Becker, E.W., 2007. Micro-algae as a source of protein. Biotechnol. Adv., 25: 207-210.
Benaiges, M.D., J. Lopez-Santin and C. Sola, 1989. Partial purification of 5-phosphodiesterase activity from barley rootlets. Enzyme Microbiol. Technol., 11: 444-451.
Bhalla, T.C., N.N. Sharma and M. Sharma, 2007. Production of Metabolites, Industrial Enzymes, Amino Acid, Organic Acids, Antibiotics, Vitamins and Single Cell Proteins. National Science Digital Library, India.
Callihan, C.D. and J.E. Clemmer, 1979. Biomass from Cellulosic Materials. In: Microbial Biomass-economic Microbiology, Rose, A.H. (Ed.). Academic Press, London, pp: 271-273.
Chandrani-Wijeyaratne, S. and A.N. Tayathilake, 2000. Characteristics of two yeast strain (Candida tropicalis) isolated from Caryota urens (Khitul) toddy for single cell protein production. J. Natl. Sci. Found. Sri Lanka, 28: 79-86.
Cooney, C.L., 1986. Continuous Culture: A Tool for Research, Development and Production. In: Perspectives in Biotechnology and Applied Microbiology, Alanl, D.I. and M. Moo-Young (Eds.). Elsevier Applied Science, London, pp: 271-276.
Cordero, B. and D. Voltolina, 1997. Viability of mass algal cultures preserved by freezing and freeze-drying. Aquac. Eng., 16: 205-211.
Curran, D.A., B.J. Tepper and T.J. Montville, 1990. Use of bicarbonates for microbial control and improved water-binding capacity in cod fillets. J. Food Sci., 55: 1564-1566.
Direct Link |
Damodaran, S. and J.E. Kinsella, 1983. The use of chaotropic salts for separation of ribonucleic acids and proteins from yeast nucleo-proteins. Biotechnol. Bioeng., 25: 761-770.
PubMed | Direct Link |
Engler, C.R., 1985. Disruption of Microbial Cells. In: Comprehensive Biotechnology, Moo-Young, M. and C.L. Cooney (Eds.). Pegamon Press, Oxford, UK., pp: 305-324.
Ferrianti, M.P. and A. Fiechter, 1983. Production and Feeding of Single Cell Protein. Applied Science Publications, London, ISBN: 085334231, pp: 201.
Forage, A.J. and R.C. Righelato, 1979. Biomass from Carbohydrates. In: Microbial Biomass-economic Microbiology, Rose, A.H. (Ed.). Academic Press, New York, pp: 289-292.
Hedenskog, G. and H. Morgen, 1973. Some methods for processing of single cell protein. Biotechnol. Bioeng., 15: 129-142.
PubMed | Direct Link |
Huang, Y.T. and J.E. Kinsella, 1986. Functional properties of phosphorylated yeast protein: Solubility, water-holding capacity and viscosity. J. Agric. Food Chem., 344: 670-674.
CrossRef | Direct Link |
Humphrey, A.E., 1975. Product Outlook and Technical Feasibility of SCP. In: Single Cell Protein II, Steven, R., S.R. Tannenbaum and D.I.C. Wang (Eds.). MIT Press, Cambridge, MA., pp: 1-23.
Kim, J.K. and H.Y. Chung, 2001. Preservation of manipulated yeast diet. Aquac. Int., 9: 171-181.
CrossRef | Direct Link |
Kim, J.K., K.T. Tak and J.H. Moon, 1998. A continuous fermentation of Kluyveromyces fragilis for the production of a highly nutritious protein diet. Aquac. Eng., 18: 41-49.
Kunhi, A.A.M. and M.R.R. Rao, 1995. The utility of a fungal ribonuclease for reducing the nucleic acid content of permeabilized yeast cells. Food Biotechnol., 9: 13-28.
Direct Link |
Lee, B.H., 1996. Fundamentals of Food Biotechnolory. Wiley-VCH, New York.
Mahasneh, I.A., 1997. Production of single cell protein from five strains of the microalga Chlorella sp. (Chlorophyta). Cytobiosciences, 90: 153-161.
Maul, S.B., A.J. Sinskey and S.R. Tannenbaum, 1970. New process for reducing the nucleic acid content of yeast. Nature, 228: 181-181.
CrossRef | Direct Link |
Middelberg, A.P.J., 1995. Process-scale disruption of microorganisms. Biotechnol. Adv., 13: 491-551.
Miller, B.M. and W. Litsky, 1976. Single Cell Protein in Industrial Microbiology. McGrow-Hill Book Co., New York.
Moebus, O. and M. Teuber, 1983. General Aspects of Production of Biomass by Yeast Fermentation from Whey and Permeate. In: Production and Feeding of Single Cell Protein, Ferranti, M.P. and A. Fiechter (Eds.). Applied Science Publishers, London, pp: 102-107.
Moulin, G., B. Malige and P. Galzy, 1983. Balanced flora of an industrial fermenter: Production of yeast from whey. J. Dairy Sci., 66: 21-28.
Direct Link |
Omar, S. and S. Sabry, 1991. Microbial biomass and protein production from whey. J. Islamic World Acad. Sci., 4: 170-172.
Direct Link |
Oura, E., 1983. Biomass from Carbohydrates. In: Biotechnology, Dellweg, H. (Ed.). Verlag Chemie GmbH, Weinheim, Germany, pp: 3-42.
Parajó, J.C., V. Santos, H. Domínguez and M. Vazquez, 1995. NH4OH-based pretreatment for improving the nutritional quality of single cell protein. Applied Biochem. Biotechnol., 55: 133-149.
CrossRef | Direct Link |
Radmer, R.J., 1996. Algal diversity and commercial algal products. Bioscience, 46: 263-270.
Direct Link |
Raja, R., S. Hemaiswarya, N. A. Kumar, S. Sridhar and R. Rengasamy, 2008. A perspective on the biotechnological potential of microalgae. Cr. Rev. Microbiol., 34: 77-88.
Direct Link |
Rasoul-Amini, S., Y. Ghasemi, M.H. Morowvat and A. Mohagheghzadeh, 2009. PCR amplification of 18S rRNA, single cell protein production and fatty acid evaluation of some naturally isolated microalgae. Food Chem., 116: 129-136.
Renaud, S.M., D.L. Parry and L.V. Thinh, 1994. Microalgae for use in tropical aquaculture I: Gross chemical and fatty acid composition of twelve species of microalgae from the Northern Territory, Australia. J. Applied Phycol., 6: 337-345.
CrossRef | Direct Link |
Richmond, A., 2004. Handbook of Microalgal Culture: Biotechnology and Applied Phycology. Blackwell, Oxford, UK.
Rivière, J., 1977. Microbial Proteins. In: Industrial Applications of Microbiology, Moss, M.O. and J.E. Smith (Eds.). Surrey University Press, London, pp: 105-149.
Roth, F.X., 1980. Micro-organisms as a source of protein for animal nutrition. Anim. Res. Dev., 12: 7-19.
Rumsey, G.L., S.G. Hughes and J.L. Kinsella, 1990. Use of dietary yeast Saccharomyces cerevisiae nitrogen by lake trout. J. World Aquac. Soc., 21: 205-209.
CrossRef | Direct Link |
Scrimshaw, N.S. and J.C. Dillen, 1977. Single Cell Protein as Food and Feed. In: Single Cell Protein-safety for Animal and Human Feeding, Garattini, S., S. Paglialunga and N.S. Scrimshaw (Eds.). Pergamon Press, Oxford, UK., pp: 171-173.
Sinclair, C.G. and D. Cantero, 1990. Fermentation Modelling. In: Fermentation: A Practical Approach, McNeil, B. and L.M. Harvey (Eds.). IRL Press, Oxford, UK., pp: 65-113.
Solomons, G.L., 1985. Production of Biomass by Filamentous Fungi. In: Comprehensive Biotechnology, Blanch, H.W., S. Drew and D.I.C Wand (Eds.). Pergamon Press, Oxford, UK., pp: 483-505.
Stanbury, P.F., A. Whitaker and S.J. Hall, 2000. Principles of Fermentation Technology. 2nd Edn., Elsevier Applied Science Publishers, New York, USA.
Steinkraus, K.H., 1986. Microbial Biomass Protein Grown on Edible Substrates: The Indigenous Fermented Foods. In: Microbial Biomass Protein, Moo-Young, M. and K.F. Gregory (Eds.). Elsevier Applied Science, London, pp: 35-45.
Trinci, A.P.J., 1992. Myco-protein: A twenty-year overnight success story. Mycol. Res., 96: 1-13.
Zadrazil, F. and A.K. Puniya, 1995. Studies on effect of particle size on solid state fermentation of sugar cane bagasse into animal feed using white rot fungi. Bioresour. Technol., 54: 85-87.
Direct Link |