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SINGLE CELL PROTEIN (SCP)

Single cell protein (SCP) refers to the microbial cells or total protein extracted from pure microbial cell culture which can be used as protein supplement for humans or animals. The word SCP is considered to be appropriate, since most of the microorganisms grow as single or filamentous individuals. If the SCP is suitable for human consumption, it is considered as food grade. SCP is regarded as feed grade, when it is used as animal feed supplement, but not suitable for human consumption. SCP broadly refers to the microbial biomass or protein extract used as food or feed additive. Besides high protein content (about 60-80% of dry cell weight), SCP also contains fats, carbohydrates, nucleic acids, vitamins and minerals. Another advantage with SCP is that it is rich in certain essential amino acids (lysine, methionine) which are usually limiting in most plant and animal foods. Thus, SCP is of high nutritional value for human or animal consumption.

Micro-organisms used for SCP production:

A large number of different species of algae, fungi, yeasts and bacteria are used for SCP production. The micro organisms are selected on the basis of following criteria:

Ability to utilize a range of carbon and nitrogen sources for SCP production. Moderate growth conditions, tolerance to pH, temperature and mineral concentration.

Reasonable protein yield. Resistance against bacteriophages. Non-toxicity and non-pathogenicity. Easy separability of cell mass in the final stages. Acceptable nutritive value of the cell biomass, etc.

Microbial biomass for food and feed:

ALGAE:
Members of the genera Chlorella, Scenedesmus and Spirulina are generally grown in ponds or tanks. They use CO2 and sunlight as substrate, which are without any cost. Generally, the limiting factor in their large scale production is illumination. Algal SCP has about 60% crude protein, which is generally good in amino acid composition except for some deficiency in sulphur containing amino acids. They are suitable for animal feed as protein rich supplements.

Disadvantages: 1) they are rich in chlorophyll content, which is not desirable for
human use. 2) They produce only low cell density, e.g., 1-2 g dry weight/l. 3) There is a serious risk of contamination. 4) Costly recovery methods have to be used for unicellular algae.

YEASTS:
Members of Saccharomyces, Candida and Torulopsis have been widely stidied for SCP production and those of the first two genera are used for some commercial processes using various substrates. E.g., Saccharomyces cerevisiae, S. lipolytica, S. fibuligera, methylotrophic yeast Pichia pastoris. The SCP has 55-60% crude protein, which has good amino acid balance, except for a deficiency in S-containing amino acids. It is usually very rich in B-group of vitamins. The SCP is used both for human food and animal feed supplementation. The risk of bacterial contamination is low and recovery by continuous centrifugation is easy.

Disadvantages: 1) slower growth rates than the fastest growing bacteria.

2) High nucleic acid content (up to 15%), which needs to be reduced. 3) Deficiency of sulphur containing amino acids.

Filamentous fungi:
Filamentous fungi have been used to produce SCP mainly from polysaccharide hydrolysates, e.g., starch hydrolysates, sulphite liquor from wood pulp industries, etc. Different fungi are Aspergillus, Fusarium, Rhizopus, Trichoderma. They have a wide range of substrate utilization, simple nutritional requirements and the ability to withstand variable pH, temperature and mineral concentrations. These are usually grown as submerged cultures in which they grow as yeastlike cells, in filamentous form or in pellets. They have crude protein content of 50-55%; the protein is low in Scontaining amino acids, but otherwise is excellent in amino acid composition. The recovery of filamentous and pellet forms is rather easy by filtration.

Disadvantages:

1) they have slower growth rates than bacteria and yeast, but

some microfungi may be comparable to yeast, 2) Contamination by yeast may be frequent if sterility is not maintained. 3) They have high nucleic acid content (up to 15% RNA), which must be reduced. 4) The strains have to be thoroughly evaluated for mycotoxins production.

BACTERIA:
A large number of bacterial species have been evaluated for SCP production. E.g., Methylophilus, Bacillus, Lactobacillus, Pseudomonas, Aeromonas. Bacteria too can utilize a wide range of carbon and nitrogen sources and have a very high growth rate. SCP has very high crude protein (over 80%) of good amino acid composition, although in some cases a small deficit of sulphur-containing amino acids may be encountered.

Disadvantages: 1) high nucleic acid content, especially, RNA content, which must
be reduced. 2) Risk of contamination by pathogenic bacteria is considerable. 3) Recovery of cells is problematic. 4) Careful evaluation for endotoxin production is essential particularly when gramnegative bacteria are used.

Nucleic acids and their removal:

Degradation of nucleic acids produces uric acid, which may accumulate to damaging levels in humans since they do not posses uricase activity.

Methods of removal/reduction:
1) Activation of endogenous RNases by, usually, a brief heat treatment, e.g., 20 min at 64C reduces RNA from 10% to 1% of dry weight in Fusarium graminearum SCP. 2) Alkaline hydrolysis. 3) Chemical extraction.

Production of SCP:
The process of SCP production from any microorganism or substrate would have the following basic steps: Provision of a carbon source; it may need physical and chemical pretreatments. In addition to the carbon source, sources of nitrogen, phosphorus and other nutrients needed to support optimal growth of the selected microorganism. Prevention of contamination by maintaining sterile conditions. The medium components may be heated or sterilized by filtration and fermentation equipments may be sterilized. The selected microorganism is inoculated in a pure state. SCP processes are highly aerobic. Therefore, adequate aeration must be provided. In addition, cooling is necessary as considerable heat is generated. The microbial biomass is recovered from the medium. Processing of the biomass for enhancing its usefulness and storability.

Biomass production is ordinarily carried out in the continuous mode to maximize yields and economic returns.

Factors affecting SCP production:

SCP production is definitely a complex and costly process. The production is greatly affected by the kind of micro-organisms chosen and the facility available for harnessing the final product. The factors like pH, temperature, humidity, O2 requirement, C & N sources and above all the fermenter design play an important role in SCP production. A problem associated with biomass production is the heat generation during the microbial growth of thick cultures which requires cooling equipments, adding to the cost of the product. The use of thermophilic

microorganisms in SCP production is recommended due to their ability to grow effectively at elevated temperature. The efficiency of chemical process increases at high temperatures. Normally, the optimum pH for SCP production is between 6.0-7.2 but the yeasts and fungi grow best in the pH range of 4.5-7.2. SCP production is greatly affected by the type and quantity of carbohydrate or organic nitrogen substrates used in the production medium.

Use of SCP as food, feed or as an ingredient:

Usually the SCPs produced on natural substrates are considered safer than those produced on hydrocarbons and related substrates.

v SCP has potential application in animal feed, human food and as an ingredient e.g. Protein concentrates and isolates are used in supplementing the human food. v Some of the commercial SCPs TOPARINA, LIQUIPRON, PRUTEEN have been considered safe for their use in animal feed, after undergoing extensive nutritional and toxicological testing. v The products from yeasts are used as protein and vitamin supplement in canned refreshments, bakery products, cheese, sausages, beverages etc. v Microbial protein is also used as flavour enhancer and in improving the texture of the baked foods.

Advantages of SCP:
The SCP is rich in high quality protein and is rather poor in fats, which is rather desirable. It can be produced all the year round and is not dependent on the climate. The microbes are very fast growing and produce large quantities of SCP from relatively very small area of land. They use low cost substrates and, in some cases, such substrates, which are being wasted and causing pollution to the environment. When the substrate used for SCP process is a source of pollution, SCP production helps reduce pollution. Strains having high biomass yields and a desirable amino acid composition can be easily selected or produced by genetic engineering. Some SCPs are good sources of vitamins, particularly B-group of vitamins, as well, e.g., yeasts and mushrooms. At present, SCP appears to be the only feasible approach to bridge the gap between requirement and supply of proteins. The culture conditions and the fermentation processes are very simple. Micro-organisms manipulations. can be easily handled, and subjected to genetic

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Limitations of SCP:
1. The nucleic acid content of microbial biomass is very high (4-6% in algae; 10-15% in bacteria; 5-10% in yeast). This is highly hazardous, since humans have limited capacity to degrade nucleic acids. 2. The presence of carcinogenic and other toxic substances is often observed in association with SCP. These include the hydrocarbons, heavy metals, mycotoxins and some contaminants. 3. There is a possibility of contamination of pathogenic micro-organisms in the SCP. 4. The digestion of microbial cells is rather slow. This is frequently associated with indigestion and allergic reactions in individuals. 5. Food grade production of SCP is more expensive than some other sources of proteins e.g. soy meal. This mainly depends on the cost of raw materials.

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Biomass recovery:
The general approaches for the recovery of biomass are as follows: (1) bacteria-flocculation and floatation combined with centrifugation, (2) yeastcentrifugation, (3) filamentous organisms-filtration.

It is important to reduce water as much as possible to reduce drying costs. Sun-drying is cheap but it reduces the quality of SCP. Since recovery processes are not done under sterile conditions, adequate hygiene must be maintained and care should be taken to prevent growth of undesirable micro-organisms. Heat treatments are used during the final stages of harvesting to inactivate heat-sensitive organisms and to reduce RNA content. The cell walls must be broken to enhance the nutritional value of SCP. The biomass may be further processed or even the protein may be isolated and purified.

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