Applications of Biotech PDF

Application of Biotechnology Microbial Applications Partha Guha Lecturer Dept. of Biochemistry & Biotechnology Ref:...

Application of Biotechnology Microbial Applications Partha Guha Lecturer Dept. of Biochemistry & Biotechnology Ref: A Textbook of Biotechnology by R.C. Dubey Chapter-17: Microbial products (primary and secondary metabolites). \\Vitamin and Organic acid part. Chapter-18: Single cell proteins and mycoproteins.  Importance of Microorganisms In recent years, a revolutionary success has been made for the production of alcohols, vitamins, organic acids, enzymes, antibiotics etc. on large scale. All of this is possible by controlling the metabolic activity of different microorganisms. It is the inherited ability of the microbes that they utilize a variety of organic substances and result in a wide range of products through metabolic processes. Importance of Microorganisms The microbial cell culture acts as a bioreactor or biofactory and converts the raw materials (different substrates) into various products during specific time. Owing to presence of biochemical diversity, microorganisms have been commercially exploited by fermentation industry. Metabolites Intracellular Extracellular Substrate biochemical macromolecules reaction Biomass Requirements for Microbial Culture Microorganisms need optimum conditions for growing in the culture media: pH Temperature Humidity Atmospheric pressure Solvent The microbes utilize the nutrients at optimum conditions and convert into desired products. Utilizing a major amount of substrate the microbes also increase the biomass and growth multiplication. Fermentation Process of Microbes Fermentation is a metabolic process in which microorganisms like bacteria, yeast, or fungi convert organic compounds, typically carbohydrates, into simpler compounds such as acids, gases, or alcohols under anaerobic conditions (without oxygen). The fermentation is completely an anaerobic process. Here organisms generate ATP (energy) through substrate-level phosphorylation during glycolysis. Depending on the type of fermentation and organism involved the common end products include: ethanol, lactic acid, acetic acid, and gases like carbon dioxide. Fermentation Process of Microbes  Vitamin Production All phototropic microorganisms are capable of synthesizing vitamins and other growth stimulating compounds for their vegetative growth by using the chemical constituents from the culture medium. When synthesis of this compounds exceed beyond their requirement, it accumulates in the culture; there from it is recovered. Now-a-days, commercial exploitation of such microbes is being done which synthesize vitamins on large scale under different culture condition. Vitamin-B12 (Cobalamin) Production This water-soluble vitamin is produced through microbial fermentation. It's used in the food, feed, healthcare, and medical industries. In nature, Vit-B12 is synthesized by microbes. For industrial production of vitamin B12 a number of bacteria like streptomycetes are used. The amount of B12 produced by them has been estimated about 20mg/L Organic Acid Production A microorganism grows in broth of the fermenter and during its growth, it produce organic acids. The organic acids are produced through the metabolism of carbohydrates. The organic acid accumulate in broth of fermenter, wherefrom they are separated and purified. Citric Acid Production Citric acid is an organic compound. It is a colorless weak organic acid. It occurs naturally in citrus fruits. Chemically citric acid was synthesized from the glycerol. Microbes can also produce citric acid for their metabolic activity. Commercially Aspergillus niger is used for producing citric acid. Citric Acid Production Culture medium: Carbohydrates, KH2PO4, MgSO4. 7H20,n Cu, Fe, pH:1.2-2.0 Addition of Aspergillus niger (A. niger) Fermentation 27-35⁰c over night to 4-16 days Fermentation Broth Add lime (CaOH2) Calcium citrate Add H2SO4 Calcium sulphate+ Citric acid Citric acid Filter Anhydrous crystalline Citric acid chemical Vacuum evaporator Crystals (commercially available) Of Citric acid Citric acid crystals + Citric acid Centrifuge  Citric Acid Production The culture medium contains carbohydrate, KH2PO4, MgSO4. 7H20 and few other trace elements. Solution of carbs are high test cane syrup. After culture medium preparation, A. niger strains (commercial) were added or cultured in the culture medium. After fermentation process the fermentation broth is collected because it acts as a source of citric acid. Lime (CaOH2) is added to the broth to allow precipitation of citric acid in the form of Calcium citrate. Again the precipitate is treated with H2SO4 to precipitate insoluble calcium sulfate. Then it is filtered and the filtrate contain citric acids. The citric acid is purified by passing through column of carbon granules (Treated with heat and washed with HCl). The citric acid is concentrated under vacuum to form crystals. The crystals are further separated by centrifugation and made commercially available in the form of anhydrous crystalline salt.  Single Cell Protein (SCP) The dried cells of microorganisms like algae, bacteria, fungi used as food or feed or “Microbial protein”. Since the ancient times a number of microorganisms have been used as a part of diet. Like, yeast for bread, fermented milk and cheese produced by lactic acid bacteria. The term “Microbial protein” was replaced by a new term “Single cell protein” or SCP. Advantages of producing Single Cell Protein (SCP) 1. Rapid succession of generations (Algae: 2-6hrs; yeast: 1-3hrs; bacteria: 0.5-2hrs). 2. Easily modified by genetic engineering. 3. High protein content of 43-85% in total dry mass. 4. Broad spectrum of raw material used for production which also include waste product. 5. Consistent quality of SCPs, not dependent on climate in determinable amount, low land requirement, ecologically beneficial. Single Cell Protein (SCP) Substrate used for SCP Production A variety of substrate can be used for SCP production. However availability of necessary substrate, its biological and economic importance are needed to be considered for the production of SCPs. Sugar: Sugarcane, sugarbeet Starch: Grains, Potatoes Lignocellulose from woody plants Organic wastes from certain industries and are rich in aromatic compounds or hydrocarbons. SCP Product of Spirulina Spirulina is a blue-green algae that's a popular source of SCP. It's a filamentous, photosynthetic bacteria that grows in alkaline lakes and salt or fresh water. Spirulina is often used in supplementary diets for malnourished children, sportsmen's diets, and baby foods. It can also be sprinkled over soft, moist food. Spirulina is rich in nutrients, including protein, vitamins, minerals, and essential fatty acids. It's also known for its health benefits, such as boosting the immune system, reducing inflammation, and providing antioxidant benefits. Benefits of Spirulina Being a filamentous algae, Spirulina can be harvested by simple and less expensive method. Filaments of Spirulina float on water surface due to the presence of gas vacuoles. No problem in harvesting. There is least chance of contamination in a growth tank as it grows at high alkaline pH:8-11. Heat drying is enough for spirulina at it has a thin cell wall. Spirulina is highly digestive (85-95%) due to thin cell wall and low nucleic content. It contains high level of digestive proteins (62-72%), vitamins, amino acids and other nutrients. Bacterial Biomass Production Microbial biomass is the total mass of living microorganisms in soil, including bacteria, fungi, algae, and protozoa. Bacterial biomass is the total number of bacteria in the microbial biomass, which is the sum of all microorganisms in a given area. Microbial biomass is a key indicator of soil quality and a vital ecological parameter. It's also a measure of disease suppression, as more microorganisms compete with pathogens for food and space. Biomass formation is also very much important for producing large amount of SCP. Biomass production basically means increasing the number of a specific microorganism in a specific container or apparatus by providing necessary nutrients and time. Bacterial Biomass Production Some steps required for bacterial biomass production, Step 1: Supply of nutrient substrate Step 2: Formulation of suitable medium Step 3: Multiplication of microorganisms through fermentation Step 4: Separation of cellular substances from left over medium Step 5: Further treatment to kill and dry bacterial biomass. Biotechnology in Service of Environment  Biotechnology in Service of Environment Biotechnology plays a critical role in addressing environmental challenges by providing innovative solutions for pollution control, waste disposal, energy production, and environmental restoration. Pollution control Biotechnology offers sustainable approaches to reduce and manage pollution: 1. Biofilters and Bioreactors: Biofiltration is a pollution control technique using a bioreactor containing microbes to capture and biologically degrade pollutants. Common uses include processing waste water and air for capturing harmful chemicals. 2. Biosensor: A biosensor uses a biological entity (i.e. bacteria) to monitor levels of certain chemicals OR uses chemicals to monitor levels of certain biological entities (i.e. pathogens). Pollution control 3. Degradation of Plastics: Development of enzymes and microbes like bacteria, algae and fungi that degrade plastics into environmentally friendly by-products. 4. Treatment of Oil Spills: Microbes such as Pseudomonas species are used to break down hydrocarbons in oil spills.  Waste Disposal and Biogas Production Biotechnology aids in converting waste into useful products while reducing its environmental impact. 1. Biogas Production: Organic waste, including agricultural residues and animal manure, is anaerobically digested by microbes to produce methane-rich biogas. The process generates renewable energy while reducing greenhouse gas emissions. Moreover, microbial decomposition of organic waste produces nutrient-rich compost for agriculture. 2. Industrial Wastewater Treatment: Microbial consortia degrade organic pollutants in industrial effluents, making the water reusable. Phytoremediation and Bioremediation 1. Phytoremediation: Phytoremediation is a plant-based method that uses plants to clean up contaminated soil, water, and air. Simply it is the use of plants to remove, stabilize, or detoxify contaminants from soil and water. Storing contaminants: Plants can store contaminants in their roots, stems, or leaves. Converting contaminants: Plants can convert contaminants into less harmful chemicals or vapors. Sorbing contaminants: Plants can stick contaminants onto their roots, where microbes break them down. Examples: Sunflowers for heavy metal absorption in polluted soils. Duckweed to remove excess nutrients from water bodies. Phytoremediation and Bioremediation 2. Bioremediation: Bioremediation is a process that employs the use of living organisms such as microbes and bacteria to decontaminate affected areas. It's used in the removal of contaminants, pollutants, and toxins from soil, water, and other environments. Bioremediation is used to clean up oil spills or contaminated groundwater. Bioremediation can be accomplished "in situ" at the site of the contamination or "ex situ" away from the site. Examples: Bacillus species for pesticide degradation. Geobacter species for uranium and heavy metal cleanup Phytoremediation and Bioremediation 2. Bioremediation: Bioremediation relies on stimulating the growth of certain microbes that utilize contaminants including oil, solvents, and pesticides for sources of food and energy. These microbes convert contaminants into small amounts of water as well as harmless gases such as carbon dioxide. Bioremediation requires a combination of the right temperature, nutrients, and foods. The absence of these elements can prolong the cleanup of contaminants. Thank You!