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Library of Congress Cataloging-in-Publication Data
ISBN: 978-1-119-46026-8
Ritika Joshi, Vinay Sharma and Arindam Kuila*
Bioscience & Biotechnology Department, Banasthali University, Rajasthan, India
*Corresponding author: arindammcb@gmail.com
This chapter deals with the current status and future prospects of the fermentation technology (FT). It discusses the different types of fermentation processes (solid-state and submerged fermentation) as well as the different types of enzyme and antibiotics production by FT. In addition, various industrial applications (enzyme production, organic acid production, biofuel production, etc.) of solid-state fermentation are also discussed. Also discussed are the future prospects of FT with regard to enhanced value product development.
Keywords: Fermentation technology, solid-state fermentation, enzyme production, biofuel production
Fermentation technology is defined as field that involves the use of microbial enzymes for production of compounds that have application within the energy production, material, pharmaceutical industries, chemical, and food industries [1].
It appears naturally in various foods. The human beings are using it from the ancient times for preservation and organoleptic properties of food. It is a well-established technology of the ancient time used for food preservation, production of bread, beer, vinegar, yogurt, cheese, and wine. From time to time, it has got refined and diversified [2].
It is the biological process in which various microorganisms such as yeast, bacteria, and fungi are involved in the conversion of complex substrate into simple compounds which are useful to humans (enzymes production, metabolites, biomass, recombinant technology, and biotransformation product) on industrial scale. Organic acid and alcohol are the main products of fermentation. In this process, there is liberation of secondary metabolites like antibiotics, enzymes, and growth factors [3, 4].
They acquire biological activity so they are also known as bioactive compounds. These compounds contain plant and food constituents in small amount which are very nutritional. Various bioactive compounds consist of secondary metabolites, for example phenolic compounds, growth factors, food pigments, antibiotics, mycotoxins, and alkaloids [5, 6]. The constituent of phenolic compounds are flavonoids, tannins, and phenolic acids. Flavanones, flavonols, flavones, anthocyanidins, and isoflavones are some major classes of flavonoids. Flavonoid comprises largest collection of plant phenolics where most of them are naturally occurring compounds [7].
According to their diverse perspectives, food and beverage are used in modern industrial fermentation processes. On the bases on different parameters such as environmental parameters and organisms required for fermentation, these techniques have become more advanced.
Generally, bioreactor is required in the middle of this process which can be arranged on the basis of their feeding of the batch, continuous and fed-batch fermentation, immobilization process. In the presence of the available amount of oxygen, mixing of substrate take place in single and mixed culture in submerged fermentation (SmF) [8].
Solid-state (or substrate) fermentation (SSF) are define as fermentation that place in solid supporting, non-specific, natural state, and low moisture content. In this process, substrates such as nutrient rich waste can be reused. Bran, bagasses, and paper pulp are the solid substrates used in SSF. Since the process is slow the fermentation of substrate takes long time. So, the discharge of the nutrients is in controlled manner. It requires less moisture content so it is the best fermentation technology used for fungi and microorganism. However, this process is not applicable for bacteria because this fermentation cannot be used for organism that requires high water condition [9].
In SmF, microorganism required a controlled atmosphere for proficient manufacture of good quality end products; attain optimum productivity and high yield.
Batch, fed-batch, or continuous modes are used in industrial bioreactors for the production of different type of microorganism in broad range [8].
For the manufacture of alcoholic beverages (whisky, beer, brandy, rum, and wine), preservatives or acidifiers (lactic acids, citric, and vinegar) are used in food industry and for flavor enhancers (monosodium glutamate) or sweeteners (aspartate) amino acid are used in submerged batch cultivation.
In this part, there are different ways of submerged cultivation using microorganisms in bioreactors. Here we have discussed briefly about typical features and advantages and faults of each fermentation methods are displayed. Lastly, the production of microorganism in liquid medium in various type of food industrial product has been determined as the most important application for continuous, batch, and fed-batch cultivation.
Batch culture is a closed system which works under aseptic condition. In these cultivations, inoculums, nutrients, and medium are mixed in the bioreactor in which the volume of the culture broth remains constant.
It is very important to select a good substrate as the product of fermentation extremely varies. This technique is used for optimization of every substrate. This is mainly due to the cause that microorganism reacts in different way in every substrate.
The rate of consumption of different nutrient vary in every substrate, and so that their productivity. Some commonly used substrates in SSF are rice straw, vegetable waste, wheat bran, fruit bagasse, synthetic media, and paper pulp. Liquid media, molasses, waste water, vegetable juices, and soluble sugar are common substrates used in SmF to extract bioactive compounds.
Enzymes [10], antioxidants [11], antibiotics [12], biosurfactants [13], and pigments [14] are variety of bioactive compounds which are extracted using fermentation.
Enzyme cultivation is the most important technique for the manufacturing of different enzymes.
When fermentation on appropriate substrates is done, both fungus and bacterial microbes are required for the precious collection of enzyme. Enzyme production can be together performed by submerged and SSF. Bacterial enzyme production commonly implies SmF method because it requires high water potential [15]. In fungus, where less water potential is required, SSF method is applied [16].
In the world, 75% of the industries are using SmF for the production of enzymes. The major reason of using SSF is that it does not support genetically modified organisms (gmo) to the extent to which SmF does, so we prefer SmF rather than SSF.
One more reason of using SmF is that it has lack of paraphernalia as related to the cultivation of variety of enzymes using SSF. The microorganism is dissimilar in SmF and SSF by the detailed metabolism display that’s way this is highly critical process. Here, influx of nutrients and efflux of waste substance is carried out in different metabolic parameters of cultivation. Some small variation from the particular parameters will affect the undesirable product.
Cellulose, amylase, xylanase, and L-asparaginase are some well know enzymes produced from bacteria. Previously we have thought that SmF is one of the best ways to produce enzyme from bacteria. Current studies have shown that for bacterial enzyme production SSF is more capable than SmF. The most important explanation can be given by metabolic differences. In SmF system, lowering of enzyme activity and production efficiency is done by gathering of different intermediate metabolites.
Numerous genus of fungus, Aspergillus, has been isolated from this process which is industrially important for the production of enzyme. This fungus has been a well-known model of microorganism for the production of fungus enzyme [17]. Aspergillus is one of the largest sources of fungal enzyme. The common difference between SSF and SmF are straight lying on the productivity of the fungus [17]. Using SmF, phytase is extracted from Thermoascusauranticus [18].
The most important extract from microorganism using fermentation technology is antibiotics. It is a bioactive compound. Penicillin from Penicillium notatum is the first antibiotic produced from fermentation. It was completed in 1940s using SSF and SmF but today P. chrysogenum isolates are higher yielding producers [19]. Aminocillins, Carbapencins, Monobactams, Cephalosporins and Penicillins together they are known as β-lactam antibiotics [19]. Some other antibiotics like Tetracyclin, Streptomycin, Cyclosporin, Cephalosporin and Surfactin are manufactured from this process. Streptomyces clavuligerus, Nocardialactamdurans, and Streptomyces cattleya produces Cephamycin C from sunflower cake and cotton-de-oiled cake in which wheat raw is supplemented in SSF system as substrates for manufacturing Cephamycin C. In SSF, penicillin was produced by actinomycetes and fungi in mixed cultures.
In current time, the growth of proper substrates has led to the widespread use of SSF more than SmF. On the other hand, some results show that several microbial stains are extra suitable to SSF and others are more suitable for SmF. Thus, this technology is determined on the bases of microorganism that is being used for production. Recently, it has been studied that several antibiotics produced through SSF are more constant and high in quantity than SmF.
This is associated to minor production of bioactive substance that are intermediary compounds in SSF. However, the characteristics of the substrate material and their quality make SSF implementation limited. Due to this property, it is compulsory to check the production ability of different substrates earlier than optimization of the fermentation process.
Typically, in the beginning of batch cultivation, the bioreactors are filled with sterilized medium and the quantity of viable cell is known which is inoculated in the bioreactor. It is beneficial for the construction of biomass (Baker’s yeasts) and primary metabolites (lactic acid, citric acid, acetic acid or ethanol production).
In food industries, organic acids used as preservatives or acidifiers(lactic acids, citric acids, and acetic acids), alcoholic beverages (wine, beer, and distilled spirits i.e. brandy, whisky, and rum) and sweeteners (e.g., aspartate) or amino acids used as flavoring agents (e.g., monosodium glutamate)are the various product manufactured by submerged batch cultivation.
Fermentation of whisky is taken as a good example, the manufacturing of distilled spirits are made from wood or stainless steel and it is made in simple cylindrical vessels known as wash backs.
Even it is very difficult to clean it but they used it, mainly in malt whisky distilleries. In this process, wort is pumped and cooled to 20 °C and inoculated with the yeast cells.
It has been found that manufacturingof citric acid has reached 1.8 × 106 tons in 2010 and about 90% of this is synthesized by the fungus Aspergillus niger from sugar containing material like sugarcane, corn, and sugar beet and food industry consumed 60% of it. We can follow surface liquid fermentation, SSF, and submerged liquid fermentation for the production of citric acid in industrial scale, however, the end predominates [24].
In fed batch cultivation, one or more nutrients are added aseptically, it is a semi-open system and the culture is supplemented step-by-step into the bioreactor at the same time the volume of the liquid culture in the bioreactor increase within this time.
The increase in productivity, enhanced yield by controlled sequential addition of nutrients, ability to achieve higher cell densities, and prolonged product synthesis are the main advantages of fed-batch over batch cultures.
Immobilized Cell Technology Active Biocatalyst also known as enzyme or microbial cell has increased the productivity of bioprocesses and it is managed through controlled contact with high concentration. Through cell immobilization or recycling by feeding strategies in high density cultures [20]. Cell immobilization mostly studied in the food and gas-liquid mass. It is done in three phase bioreactor; it requires all three phases in competent mass transfer. These bioreactor aims in the region where main process amplification can be managed through the improvement of gas-liquid mass transfer [21].
Fundamental difference between SmF and SSF
| Submerged fermentation | Solid-state (substrate) fermentation |
|---|---|
| Water cultivation medium (~95%). | Water cultivation medium is low (40–80%). |
| Liquid–gas are the two phase of the system. | Solid–liquid–gas are three phase of system. |
| Homogeneous. | Heterogeneous. |
| Low nutrient content, water soluble. | High nutrient content, water insoluble. |
| Oxygen transfer: gas–liquid. | Oxygen transfer: liquid–solid and gas–liquid. |
| Microorganism growth: liquid medium. | Microorganism growth: medium surface. |
| Only oxygen is transfer, process is not limited. | Oxygen, heat, and nutrient transfer is limited. |
| Product: soluble in the liquid phase. | Product: high concentration. |
In SSF, agriculture industrial substrates are considered the most excellent for enzyme production.
The expenditure of enzyme production by SmF is high as compared to SSF.
Approximately, all well-known microbial enzymes are produced through this process. According to research study, large amount of work has been done on the enzyme production of industrial importance like cellulases, lipase, proteases, glucoamylases, amylases, ligninases, xylanases, pectinases, and peroxidases. Thermostable enzyme xylanase by thermophilic Bacillus licheniformis has been produced from this process. Enzymes produced from this process are more thermo-stable than SmF process. It has 22- folds higher in SSF system than in SmF system.
The bacterial strain extracted from open xylan agar plate are characterized as xylanase produced from Bacillus pumilus from both the processes (submerged and SSF fermentation) [22]. Rhizopus oligosporus is used to produce acid protease from rice bran and during its production no toxin effect occurred in SSF.
Gallic acid, citric acid, fumaric acid, kojic acid, and lactic acid are various acid produced by SSF. Wheat bran, de-oiled rice bran, sugarcane, carob pods, coffee husk, kiwi fruit peels, pineapple wastes, grape pomace, and apple are some agro-industrial wastes which are very resourceful substrates for production of citric acid in SSF. For the production of citric acid from Aspergillus, pine apple waste was used as substrate [23]. Sugarcane bagasse impregnated with glucose and CaCO3 for the production of lactic acid from Rhizopus oryzaeis used.
Fungus produce secondary metabolite, gibberellic acid, in its stationary phase. Gibberellic acid production increases in SSF system. Gathering of gibberellic acid was 1.626 times greater in SSF than SmF using Gibberellafujikuroi in the production of gibberellic acid in which wheat bran is used as substrate.
Cephamycin C, Cyclosporin A, Penicillin, Neomycin, Iturin, and Cephalosporins are some common antibiotics produced from SSF. Penicillin is produced from Penicillum chrysogenum in which wheat bran and sugarcane bagasse are used as substrate under high moisture content (70%). Nocardia lactamdurans, Streptomycesclavuligerus, and Streptomyces cattleya produces Cephamycin C. In SSF, antibiotic penicillin is produced from Actinomycetes and fungi through mixed cultures.
Today, ethanol is the most extensively used biofuel. Even though it is very easier to produce ethanol using SmF, it is preferred because of low water requirement, little volumes of fermentation mash, end product protection is inhibited and less liquid water disposal, it decreases pollution problem and it is most commonly used for ethanol production because of abundant availability. Saccharomyces cerevisiae is used for ethanol production in SSF of apple pomace supplemented with ammonium sulfate in controlled fermentation. Sweet potato, rice starch, wheat flour, potato starch, and sweet sorghum are commonly used substrate.
On the bases of different mode of action, fungal agent has greater potential to act as biocontrol agents. To control mosquitoes Liagenidium giganteum is used as fungal agent. It works by encysting on their larvae. Here they use larvae as a substrate for growth.
Nicotinic acid, vitamin B12, thiamine, riboflavin, and vitamins B6 are the water soluble enzyme produced on SSF with the help of different species of Rhizophus and Klebsiella, which is well-known producer of vitamin B12.
In food industries, processing microbial enzymes are extensively used as gift to fermentation technology. Yet, it is essential to make this kind of enzyme for the future development. In recent years, various new industrial and analytical applications have been drawn out for the manufacture of new products.
Fermentation technology needs evolution and enhancement for the food and beverage industries. It aim is to humanizing higher yield and production amount by means of construction, new models, bacterial strain, and process monitoring. In these areas, they have developed some modern ideas that could show the mode of cost-effectively attractive solutions.
In SSF, the area of modern instrumentation and sensor development is commendation of process monitoring is very important.
The modern technology characterized so far include different sensor of technologies like infrared spectrometry, magnetic resonance imaging, x-rays, image analysis, and respirometry. The chief drawback is high cost, so for large-scale applications this technique is unsuitable. Algae and micro/macro algae derived food production is one of the best bioreactor design for development of large-scale photo-bioreactors and phytocultures (seaweed). The use of properly controlled ultra-sonication in bioprocesses is another potential approach to enhance the metabolic productivity.
Sono-bioreactor performance (mass transfer enhancement), their function (e.g., cross-membrane ion fluxes, stimulated sterol synthesis, altered cell morphology, and increased enzyme activity) and biocatalysts (cells and enzymes) are advantageous effects of ultrasound which can be exploited.
Its prospective in the field of food fermentation for genetic engineering is indisputable. On the basis of understanding of their diet and human gastrointestinal microbiota, food fermentation has improved the nutritional status by the balanced choice of food-fermenting microbes. In this respect, food fermentation has attributed beneficial towards health and regarded as an extension of the food digestion.
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