This document discusses various attributes required for industrial microbes to be used efficiently as cloning and expression hosts. It outlines several microbial hosts commonly used for recombinant protein production, including E. coli, B. subtilis, S. cerevisiae, and others. It also discusses various vectors that can be used for cloning DNA fragments, such as plasmids, bacteriophages, cosmids, and artificial chromosomes.
Cheese is a generic term for a diverse group of milk based food products.
Cheese consists of proteins and fat from milk, usually the milk of cows, buffalo, goats, or sheep.
It is produced by coagulation of the milk protein casein.
Biodeterioration of paper and leather ppt..ShaistaKhan60
This document discusses the biodeterioration of paper and leather. It defines biodeterioration as the breakdown of materials by microorganisms or undesirable changes caused by organisms. For paper, factors like humidity, chemicals, and microbes like fungi can cause staining, foxing, and weakening. Leather deterioration is also caused by bacteria and fungi when conditions are poor, leading to hardening, deformation, and discoloration. Preventing biodeterioration requires controlling moisture, chemicals, insects, and proper storage conditions.
Psychrophiles are cold-loving microorganisms that can survive and grow in extremely cold environments like polar regions and glaciers. They have adapted mechanisms like maintaining membrane fluidity through unsaturated fatty acids and proteins that preserve RNA structures at cold temperatures. Psychrophiles possess enzymes that remain active at low temperatures through higher alpha-helix content and compounds like trehalose that prevent protein denaturation. Their adaptations allow psychrophiles and their cold-active enzymes to have applications in various industries like detergents, food processing, and bioremediation.
Sri Paramakalyani College in Alwarkurichi, India has received A+ grade accreditation from NAAC with a GCPA of 3.39. The document discusses the industrial production of amino acids like glutamic acid and monosodium glutamate (MSG) using microorganisms. It provides details on the commercial uses of amino acids in food, pharmaceutical and chemical industries. The key aspects of large scale glutamic acid production using Corynebacterium glutamicum are also summarized, including carbon and nitrogen sources, growth factors, oxygen requirements and recovery processes.
Fermentation
Scale up of fermentation
Steps in scale up
Scale up fermentation process
Optimizing scale up of fermentation process
Rules followed while doing scale up
Studies carried out during scale up
Reference
Quorum sensing allows bacteria to coordinate gene expression based on cell population density through the use of signaling molecules. As the bacteria population grows, the concentration of these signaling molecules increases until a threshold is reached that triggers a group response. Quorum quenching disrupts quorum sensing by degrading these signaling molecules. Compounds that inhibit quorum sensing have potential applications in controlling bacterial virulence without promoting antibiotic resistance.
Process scale-up is a critical activity that enables a fermentation process achieved in research and development to operate at a commercially viable scale for manufacturing.
This document summarizes the application of computers in fermentation. It discusses the initial use of computers in the 1960s for modeling fermentation processes. Computers are now used for logging process data, analyzing the data, and controlling fermentation processes. Sensors are used to monitor important factors like temperature, pH, dissolved oxygen, and mineral/nutrient levels to provide data inputs for computer control and modeling of fermentation.
Cheese is a generic term for a diverse group of milk based food products.
Cheese consists of proteins and fat from milk, usually the milk of cows, buffalo, goats, or sheep.
It is produced by coagulation of the milk protein casein.
Biodeterioration of paper and leather ppt..ShaistaKhan60
This document discusses the biodeterioration of paper and leather. It defines biodeterioration as the breakdown of materials by microorganisms or undesirable changes caused by organisms. For paper, factors like humidity, chemicals, and microbes like fungi can cause staining, foxing, and weakening. Leather deterioration is also caused by bacteria and fungi when conditions are poor, leading to hardening, deformation, and discoloration. Preventing biodeterioration requires controlling moisture, chemicals, insects, and proper storage conditions.
Psychrophiles are cold-loving microorganisms that can survive and grow in extremely cold environments like polar regions and glaciers. They have adapted mechanisms like maintaining membrane fluidity through unsaturated fatty acids and proteins that preserve RNA structures at cold temperatures. Psychrophiles possess enzymes that remain active at low temperatures through higher alpha-helix content and compounds like trehalose that prevent protein denaturation. Their adaptations allow psychrophiles and their cold-active enzymes to have applications in various industries like detergents, food processing, and bioremediation.
Sri Paramakalyani College in Alwarkurichi, India has received A+ grade accreditation from NAAC with a GCPA of 3.39. The document discusses the industrial production of amino acids like glutamic acid and monosodium glutamate (MSG) using microorganisms. It provides details on the commercial uses of amino acids in food, pharmaceutical and chemical industries. The key aspects of large scale glutamic acid production using Corynebacterium glutamicum are also summarized, including carbon and nitrogen sources, growth factors, oxygen requirements and recovery processes.
Fermentation
Scale up of fermentation
Steps in scale up
Scale up fermentation process
Optimizing scale up of fermentation process
Rules followed while doing scale up
Studies carried out during scale up
Reference
Quorum sensing allows bacteria to coordinate gene expression based on cell population density through the use of signaling molecules. As the bacteria population grows, the concentration of these signaling molecules increases until a threshold is reached that triggers a group response. Quorum quenching disrupts quorum sensing by degrading these signaling molecules. Compounds that inhibit quorum sensing have potential applications in controlling bacterial virulence without promoting antibiotic resistance.
Process scale-up is a critical activity that enables a fermentation process achieved in research and development to operate at a commercially viable scale for manufacturing.
This document summarizes the application of computers in fermentation. It discusses the initial use of computers in the 1960s for modeling fermentation processes. Computers are now used for logging process data, analyzing the data, and controlling fermentation processes. Sensors are used to monitor important factors like temperature, pH, dissolved oxygen, and mineral/nutrient levels to provide data inputs for computer control and modeling of fermentation.
Beer is an alcoholic beverage made by fermenting grains like barley with hops and yeast. The key ingredients are barley, hops, yeast, and water. There are two main types - top fermented beer made with Saccharomyces cervisiae yeast and bottom fermented beer made with Saccharomyces carlsbergences yeast. Wine is an alcoholic beverage made by fermenting fruit juice, most commonly grape juice. The major steps in the production of beer and wine are preparation of ingredients, fermentation, aging or storage, and packaging. Microbial contamination and temperature fluctuations can affect the quality of beer and wine.
This document discusses various types of extremophiles and their adaptations to extreme environments. It describes acidophiles, alkaliphiles, thermophiles, psychrophiles and their ability to thrive in highly acidic, alkaline, hot, and cold conditions respectively. Acidophiles maintain a neutral pH inside their cells while alkaliphiles actively pump out hydroxide ions. Thermophiles have heat-stable membranes and proteins while psychrophiles can grow in temperatures as low as -15°C through various metabolic pathways. The document provides examples of extremophile organisms from all domains of life that have adapted to survive in these extreme conditions through specialized cellular mechanisms.
Bacteriophages & Its classification, cycles, therapy, and applicationsZoqiaTariq
These slides are covering multiple aspects of Bacteriophages including History
Classification
Replication
Plaque Assay
Transduction
Phage Therapy and pahge types.
This document discusses strain improvement and preservation in biotechnology. It defines a strain as a group of species with distinguishing characteristics. The main approaches to strain improvement discussed are mutant selection, recombination, and recombinant DNA technology. Mutant selection involves applying mutagens to induce beneficial mutations for traits like increased productivity. Recombination generates new combinations of genes between strains. Recombinant DNA technology transfers genes to modify metabolic activities or products. Proper strain preservation methods are also outlined, including freezing, lyophilization, and storage in glycerol or liquid nitrogen. Applications include production of vaccines, enzymes, and other industrial biomolecules.
Antimicrobial metabolites of lactic acid bacteria and its applicationDiwas Pradhan
This document discusses lactic acid bacteria and their antimicrobial metabolites. It begins with an overview of the taxonomy of lactic acid bacteria and describes some of the organic acids, bacteriocins, and other low molecular weight antimicrobial compounds they can produce. It then discusses applications of these compounds in food preservation and safety, as well as medical and veterinary uses. Specifically, it explores using purified/semi-purified bacteriocins as food additives and fermenting foods or ingredients with bacteriocin-producing cultures. Overall, the document provides information on the antimicrobial compounds produced by lactic acid bacteria and their potential applications.
Steroid transformation, bioreactor and bioprocess engineeringRitasree Sarma
1. Steroids are organic molecules containing four rings of carbon atoms that are synthesized in tissues to act as hormones, alkaloids, and vitamins.
2. Common types of steroids include sex hormones, corticosteroids, mineralocorticoids, and bile salts.
3. Microbial transformation of steroids involves enzymatic reactions that can modify steroid structures through oxidation, hydroxylation, dehydrogenation, epoxidation, and other processes. This is an attractive alternative to chemical synthesis.
Microbial Polysaccharide - Food Application - Food IndustryMUTHUGANESAN N
Polysaccharides are the carbon sources which are found in huge amount in the biosphere
used for food, pharmaceutical, and medical applications
derives from the great diversity in structural and functional properties.
xanthan, xylinan, gellan, curdlan, pullulan, dextran, scleroglucan, schizophyllan, and cyanobacterial polysaccharides
The commercial value of polysaccharides is based on its ability to modify the flow characteristics of solutions (Rheology).
They can incr viscosity and hence used as thickening and gelling agents.
Production of cellulase and it's applicationRezwana Nishat
The document discusses the production of cellulase enzymes from Aspergillus isolates and its applications. Four Aspergillus isolates were identified as good cellulase producers. One isolate, Aspergillus oryzae AKAL8, produced the highest level of cellulase over time. Crude cellulase was used for denim biostoning and was found to remove more indigo dye than bleach alone. Cellulase was also stable when combined with bleach. Finally, cellulase treatment of banana peel was able to produce cellulosic nanofibers.
The document summarizes the process of beer production. Malt is made from soaked and sprouted grains like barley. Mashing involves mixing malt with water and adjuncts like rice and corn starch. This allows enzymatic degradation of starch to sugars. The liquid produced is wort, which is separated and then fermented with yeast along with hops for flavor. Fermentation yields alcohol and carbon dioxide. The final beer is pasteurized, carbonated, and aged to improve flavor before consumption.
Bacteriocin are produced from lactic acid bacteria .
various lactic acid bacteria produces different kinds of bacteriocin .
Bacteriocin can be used as food preservative
Microbes experience various forms of stress, including chemical stress from toxic compounds, physical stress like heat, and nutrient limitations. Microbes have developed stress responses to temporarily increase their tolerance limits. Stress responses can help bacteria transition from free-living to host-invading pathogens. Osmotic stress occurs when there are sudden changes in the solute concentration around a cell, causing changes in water movement across the cell membrane. Bacteria experience osmotic stress under variable, extremely low, or extremely high osmotic pressures in their environment.
Lactic acid bacteria (LAB) such as Lactobacillus, Lactococcus, Leuconostoc, and Pediococcus are important in food fermentation processes. They produce lactic acid which preserves foods and improves safety. Lactobacillus is the largest LAB genus and includes species used in dairy, bread, meat and vegetable fermentations. Lactococcus lactis is used as a starter culture for cheeses and cultured dairy. These LAB vary in their temperature and pH preferences, as well as metabolic pathways, contributing to flavor development in fermented foods through production of organic acids, aromas, and proteolysis.
The document discusses strain improvement, which is the process of manipulating microbial strains to enhance their metabolic capacities. The main methods discussed are selection of natural variants, induced mutants, and use of recombinant technology. Key characteristics for improving strains are selecting for stability, resistance to infection/components, favorable morphology, and tolerance to low oxygen. The goal is to develop strains that can be used commercially.
This document discusses various types of fermenters used in industrial fermentation processes. It describes 7 types of fermenters: 1) Waldhof fermenter, 2) Acetators and cavitators, 3) Tower fermenter, 4) Cylindro-conical vessels, 5) Air lift fermenter, 6) Deep jet fermenter, 7) The cyclone column. For each type, it provides details on their design, operating principles, and applications. The key advantages of each fermenter type for different fermentation processes are highlighted.
Organic acids like citric acid and fumaric acid can be produced via fermentation. Citric acid is produced commercially using Aspergillus niger in surface culture with sucrose as the carbon source. Key parameters that affect citric acid production include fungal strain selection, fermentation medium composition and conditions like pH, aeration and time. Fumaric acid is produced using Rhizopus nigricans in submerged culture with molasses as the carbon source. Proper control of fermentation conditions and neutralization of the medium is important for fumaric acid production and recovery.
Halophiles (Introduction, Adaptations, Applications)Jamil Ahmad
Introduction
Halophiles are organisms that thrive in high salt concentrations.
They are a type of extremophile organisms. The name comes from the Greek word for "salt-loving".
While most halophiles are classified into the Archaea domain, there are also bacterial halophiles and some eukaryota, such as the alga Dunaliella salina or fungus Wallemia ichthyophaga
The document discusses enzymes and their industrial production. It notes that enzymes are biological catalysts that accelerate chemical reactions. Common industrial enzymes include amylases, proteases, and pectinases which are produced using fungi like Aspergillus oryzae and bacteria like Bacillus species. Enzyme production involves submerged fermentation in bioreactors or semi-solid fermentation using agricultural waste. The enzymes find applications in industries like food, textiles and detergents.
Lactobacilli- Homo and Hetero lactic acid Fermentation and its nutritive value pugazhenthi6
The document discusses lactic acid fermentation by lactic acid bacteria (LAB), specifically the genera Lactobacillus. It describes how LAB convert sugars into lactic acid through either homolactic fermentation, which produces only lactic acid, or heterolactic fermentation, which produces lactic acid as well as ethanol and carbon dioxide. Applications of homolactic fermentation include dairy products and probiotics, while heterolactic bacteria are involved in other fermentation processes.
Thermophiles are microorganisms that thrive in relatively high temperatures between 45-80°C. They are classified based on their optimal growth temperatures into thermophiles, extreme thermophiles, and hyperthermophiles. Thermophiles have adapted enzymes and proteins that allow them to function at high temperatures. They are found in geothermally heated areas like hot springs and deep sea hydrothermal vents. Cyanobacteria are a common thermophile that can photosynthesize in hot spring waters up to 70°C. Thermophiles have applications in producing thermostable enzymes for uses like PCR and detergents.
This document discusses steroid biotransformation, which is the biological modification of steroids through microbial enzymes. It describes various types of microbial transformations of steroids including hydroxylation, dehydrogenation, epoxidation, and others. Commonly transformed steroids include progesterone, cortisol, and testosterone. Microorganisms like fungi and bacteria are used in fermentation to commercially produce steroid hormones and derivatives for uses as medications. The advantages of microbial transformations include enzyme selectivity and ability to produce novel compounds, while disadvantages include potential toxicity and low chemical yields.
Industrial product derived from microbsAnbarasan D
Microbial biotechnology uses microbes to produce products and services of economic value through fermentation. Some key properties of useful microorganisms include being able to produce spores or be easily inoculated, grow rapidly at large scale in inexpensive media, and produce the desired product quickly without being pathogenic or difficult to genetically manipulate. Microbes are used industrially to produce beverages, antibiotics, organic acids, amino acids, enzymes, vitamins, organic solvents, single cell protein, steroids, pharmaceutical drugs, and dairy products. Common microorganisms used include yeasts, bacteria, actinomycetes and fungi.
Microbial biotechnology refers to using microbes like bacteria and fungi to produce useful products. Microbes can be used to produce beverages, antibiotics, organic acids, amino acids, enzymes, vitamins, organic solvents, single cell protein, pharmaceuticals, and dairy products. Useful microbes for industry must grow rapidly, produce the desired product, and not be pathogenic. Common industrial microbes include Saccharomyces cerevisiae for ethanol production and Aspergillus niger for citric acid production. Microbial biotechnology provides economically important products for food, agriculture, and medicine.
Beer is an alcoholic beverage made by fermenting grains like barley with hops and yeast. The key ingredients are barley, hops, yeast, and water. There are two main types - top fermented beer made with Saccharomyces cervisiae yeast and bottom fermented beer made with Saccharomyces carlsbergences yeast. Wine is an alcoholic beverage made by fermenting fruit juice, most commonly grape juice. The major steps in the production of beer and wine are preparation of ingredients, fermentation, aging or storage, and packaging. Microbial contamination and temperature fluctuations can affect the quality of beer and wine.
This document discusses various types of extremophiles and their adaptations to extreme environments. It describes acidophiles, alkaliphiles, thermophiles, psychrophiles and their ability to thrive in highly acidic, alkaline, hot, and cold conditions respectively. Acidophiles maintain a neutral pH inside their cells while alkaliphiles actively pump out hydroxide ions. Thermophiles have heat-stable membranes and proteins while psychrophiles can grow in temperatures as low as -15°C through various metabolic pathways. The document provides examples of extremophile organisms from all domains of life that have adapted to survive in these extreme conditions through specialized cellular mechanisms.
Bacteriophages & Its classification, cycles, therapy, and applicationsZoqiaTariq
These slides are covering multiple aspects of Bacteriophages including History
Classification
Replication
Plaque Assay
Transduction
Phage Therapy and pahge types.
This document discusses strain improvement and preservation in biotechnology. It defines a strain as a group of species with distinguishing characteristics. The main approaches to strain improvement discussed are mutant selection, recombination, and recombinant DNA technology. Mutant selection involves applying mutagens to induce beneficial mutations for traits like increased productivity. Recombination generates new combinations of genes between strains. Recombinant DNA technology transfers genes to modify metabolic activities or products. Proper strain preservation methods are also outlined, including freezing, lyophilization, and storage in glycerol or liquid nitrogen. Applications include production of vaccines, enzymes, and other industrial biomolecules.
Antimicrobial metabolites of lactic acid bacteria and its applicationDiwas Pradhan
This document discusses lactic acid bacteria and their antimicrobial metabolites. It begins with an overview of the taxonomy of lactic acid bacteria and describes some of the organic acids, bacteriocins, and other low molecular weight antimicrobial compounds they can produce. It then discusses applications of these compounds in food preservation and safety, as well as medical and veterinary uses. Specifically, it explores using purified/semi-purified bacteriocins as food additives and fermenting foods or ingredients with bacteriocin-producing cultures. Overall, the document provides information on the antimicrobial compounds produced by lactic acid bacteria and their potential applications.
Steroid transformation, bioreactor and bioprocess engineeringRitasree Sarma
1. Steroids are organic molecules containing four rings of carbon atoms that are synthesized in tissues to act as hormones, alkaloids, and vitamins.
2. Common types of steroids include sex hormones, corticosteroids, mineralocorticoids, and bile salts.
3. Microbial transformation of steroids involves enzymatic reactions that can modify steroid structures through oxidation, hydroxylation, dehydrogenation, epoxidation, and other processes. This is an attractive alternative to chemical synthesis.
Microbial Polysaccharide - Food Application - Food IndustryMUTHUGANESAN N
Polysaccharides are the carbon sources which are found in huge amount in the biosphere
used for food, pharmaceutical, and medical applications
derives from the great diversity in structural and functional properties.
xanthan, xylinan, gellan, curdlan, pullulan, dextran, scleroglucan, schizophyllan, and cyanobacterial polysaccharides
The commercial value of polysaccharides is based on its ability to modify the flow characteristics of solutions (Rheology).
They can incr viscosity and hence used as thickening and gelling agents.
Production of cellulase and it's applicationRezwana Nishat
The document discusses the production of cellulase enzymes from Aspergillus isolates and its applications. Four Aspergillus isolates were identified as good cellulase producers. One isolate, Aspergillus oryzae AKAL8, produced the highest level of cellulase over time. Crude cellulase was used for denim biostoning and was found to remove more indigo dye than bleach alone. Cellulase was also stable when combined with bleach. Finally, cellulase treatment of banana peel was able to produce cellulosic nanofibers.
The document summarizes the process of beer production. Malt is made from soaked and sprouted grains like barley. Mashing involves mixing malt with water and adjuncts like rice and corn starch. This allows enzymatic degradation of starch to sugars. The liquid produced is wort, which is separated and then fermented with yeast along with hops for flavor. Fermentation yields alcohol and carbon dioxide. The final beer is pasteurized, carbonated, and aged to improve flavor before consumption.
Bacteriocin are produced from lactic acid bacteria .
various lactic acid bacteria produces different kinds of bacteriocin .
Bacteriocin can be used as food preservative
Microbes experience various forms of stress, including chemical stress from toxic compounds, physical stress like heat, and nutrient limitations. Microbes have developed stress responses to temporarily increase their tolerance limits. Stress responses can help bacteria transition from free-living to host-invading pathogens. Osmotic stress occurs when there are sudden changes in the solute concentration around a cell, causing changes in water movement across the cell membrane. Bacteria experience osmotic stress under variable, extremely low, or extremely high osmotic pressures in their environment.
Lactic acid bacteria (LAB) such as Lactobacillus, Lactococcus, Leuconostoc, and Pediococcus are important in food fermentation processes. They produce lactic acid which preserves foods and improves safety. Lactobacillus is the largest LAB genus and includes species used in dairy, bread, meat and vegetable fermentations. Lactococcus lactis is used as a starter culture for cheeses and cultured dairy. These LAB vary in their temperature and pH preferences, as well as metabolic pathways, contributing to flavor development in fermented foods through production of organic acids, aromas, and proteolysis.
The document discusses strain improvement, which is the process of manipulating microbial strains to enhance their metabolic capacities. The main methods discussed are selection of natural variants, induced mutants, and use of recombinant technology. Key characteristics for improving strains are selecting for stability, resistance to infection/components, favorable morphology, and tolerance to low oxygen. The goal is to develop strains that can be used commercially.
This document discusses various types of fermenters used in industrial fermentation processes. It describes 7 types of fermenters: 1) Waldhof fermenter, 2) Acetators and cavitators, 3) Tower fermenter, 4) Cylindro-conical vessels, 5) Air lift fermenter, 6) Deep jet fermenter, 7) The cyclone column. For each type, it provides details on their design, operating principles, and applications. The key advantages of each fermenter type for different fermentation processes are highlighted.
Organic acids like citric acid and fumaric acid can be produced via fermentation. Citric acid is produced commercially using Aspergillus niger in surface culture with sucrose as the carbon source. Key parameters that affect citric acid production include fungal strain selection, fermentation medium composition and conditions like pH, aeration and time. Fumaric acid is produced using Rhizopus nigricans in submerged culture with molasses as the carbon source. Proper control of fermentation conditions and neutralization of the medium is important for fumaric acid production and recovery.
Halophiles (Introduction, Adaptations, Applications)Jamil Ahmad
Introduction
Halophiles are organisms that thrive in high salt concentrations.
They are a type of extremophile organisms. The name comes from the Greek word for "salt-loving".
While most halophiles are classified into the Archaea domain, there are also bacterial halophiles and some eukaryota, such as the alga Dunaliella salina or fungus Wallemia ichthyophaga
The document discusses enzymes and their industrial production. It notes that enzymes are biological catalysts that accelerate chemical reactions. Common industrial enzymes include amylases, proteases, and pectinases which are produced using fungi like Aspergillus oryzae and bacteria like Bacillus species. Enzyme production involves submerged fermentation in bioreactors or semi-solid fermentation using agricultural waste. The enzymes find applications in industries like food, textiles and detergents.
Lactobacilli- Homo and Hetero lactic acid Fermentation and its nutritive value pugazhenthi6
The document discusses lactic acid fermentation by lactic acid bacteria (LAB), specifically the genera Lactobacillus. It describes how LAB convert sugars into lactic acid through either homolactic fermentation, which produces only lactic acid, or heterolactic fermentation, which produces lactic acid as well as ethanol and carbon dioxide. Applications of homolactic fermentation include dairy products and probiotics, while heterolactic bacteria are involved in other fermentation processes.
Thermophiles are microorganisms that thrive in relatively high temperatures between 45-80°C. They are classified based on their optimal growth temperatures into thermophiles, extreme thermophiles, and hyperthermophiles. Thermophiles have adapted enzymes and proteins that allow them to function at high temperatures. They are found in geothermally heated areas like hot springs and deep sea hydrothermal vents. Cyanobacteria are a common thermophile that can photosynthesize in hot spring waters up to 70°C. Thermophiles have applications in producing thermostable enzymes for uses like PCR and detergents.
This document discusses steroid biotransformation, which is the biological modification of steroids through microbial enzymes. It describes various types of microbial transformations of steroids including hydroxylation, dehydrogenation, epoxidation, and others. Commonly transformed steroids include progesterone, cortisol, and testosterone. Microorganisms like fungi and bacteria are used in fermentation to commercially produce steroid hormones and derivatives for uses as medications. The advantages of microbial transformations include enzyme selectivity and ability to produce novel compounds, while disadvantages include potential toxicity and low chemical yields.
Industrial product derived from microbsAnbarasan D
Microbial biotechnology uses microbes to produce products and services of economic value through fermentation. Some key properties of useful microorganisms include being able to produce spores or be easily inoculated, grow rapidly at large scale in inexpensive media, and produce the desired product quickly without being pathogenic or difficult to genetically manipulate. Microbes are used industrially to produce beverages, antibiotics, organic acids, amino acids, enzymes, vitamins, organic solvents, single cell protein, steroids, pharmaceutical drugs, and dairy products. Common microorganisms used include yeasts, bacteria, actinomycetes and fungi.
Microbial biotechnology refers to using microbes like bacteria and fungi to produce useful products. Microbes can be used to produce beverages, antibiotics, organic acids, amino acids, enzymes, vitamins, organic solvents, single cell protein, pharmaceuticals, and dairy products. Useful microbes for industry must grow rapidly, produce the desired product, and not be pathogenic. Common industrial microbes include Saccharomyces cerevisiae for ethanol production and Aspergillus niger for citric acid production. Microbial biotechnology provides economically important products for food, agriculture, and medicine.
Industrially important microbes their large scale productionVibhaKumari13
The above presentation is useful for the Students who want to gain and enrich their knowledge about the large scale production of industrially important microbes and fermentation procedure.
This will mainly be helpful for Students opting Agricultural microbiology
Use of microbes in industry. Production of enzymes-General consideration-Amyl...Steffi Thomas
Industrial uses of microbes, properties of useful industrial microbes, various industrial products, production of enzymes-general consideration-amylase, catalase, peroxidase, lipase, protease, penicillinase, procedure for culturing bacteria and inoculum preparation, submerged fermentation and solid state fermentation, uses of different enzymes
Microbes play an important role in human welfare through various applications such as [1] silage production, biofuel production, and biodegradation of agricultural waste. [2] Lactic acid bacteria are important in silage production by preserving crops through controlled fermentation. [3] Yeast and bacteria can ferment sugars from plants or algae into biofuels like ethanol.
1. Microbes play a crucial role in various industrial processes by producing important products like beverages, antibiotics, organic acids, amino acids, enzymes, vitamins, biofuels, proteins, steroids, vaccines, pharmaceutical drugs, and dairy products.
2. Common industrial microbes used are yeast, bacteria, actinomycetes and fungi which produce ethanol, citric acid, antibiotics, lysine, amylase and vitamins through fermentation.
3. These microbial products have significant applications in food, fuel, chemical, pharmaceutical and other industries demonstrating microbes' valuable contributions.
This document provides an overview of a course on food and industrial microbiology. It discusses 12 ways that microbes are used industrially, including to produce beverages, antibiotics, amino acids, organic acids, vitamins, enzymes, organic solvents, single cell protein, steroids, vaccines, pharmaceutical drugs, and dairy products. Examples of specific microbes used are also provided for several applications, such as Saccharomyces cerevisiae for producing ethanol in beverages and various bacteria and fungi for producing different organic acids.
This document discusses the use of microbes in industry, focusing on the production of enzymes like amylase. It notes that industries grow microorganisms in large tanks called fermentors to produce products on a large scale. Examples of industrial microbe uses include producing alcoholic beverages through fermentation and antibiotics. The document also describes the production and uses of the enzymes amylase and beta-amylase, which are used to hydrolyze starches. It highlights that bacteria like Bacillus species are commonly used to commercially produce alpha-amylase.
Lactic acid bacteria and other microbes play important roles in food production. Lactobacillus acidophilus, L. lactis, and Streptococcus lactis in yogurt and cheese help digest milk and produce beneficial compounds. Saccharomyces cerevisiae is used to produce bread, beer, wine and other alcoholic beverages through fermentation. Microbes also have many industrial uses including producing antibiotics, organic acids, amino acids, vitamins and other chemicals. They help treat wastewater and produce biofuels and enzymes.
Microbes are widely used in industry for the production of various products. Some key applications of microbes discussed in the document include:
1. Microbes are used in the food industry to produce dairy products, beverages, and bread through fermentation processes. Yeast is commonly used to produce alcoholic beverages while lactic acid bacteria produce yogurt and cheese.
2. Microbes also produce industrial chemicals like organic acids, amino acids, antibiotics, vitamins, and enzymes through fermentation. For example, citric acid and lactic acid are produced using fungi and bacteria respectively.
3. Probiotics containing beneficial bacteria and yeast are used for their health benefits such as strengthening the immune system and digestive barrier.
Microbes, or microscopic organisms, are widely used in large-scale industrial processes. Microbes can be used to create biofertilizers or to reduce metal pollutants. Microbes can also be used to produce certain non-microbial products, such as the diabetes medication insulin, vaccines, etc. These slides will give insights into uses of microbes in production of enzymes, antibiotics, beverages, vitamins, vaccines, probiotics, etc
The document discusses the various ways microbes are beneficial to human welfare. It describes how microbes are used in household food production like idli and dhokla through fermentation. It also explains how microbes or their cells can be directly used as food and mentions some edible mushrooms. The document then discusses the industrial uses of microbes like in producing alcoholic beverages, organic acids, vitamins, antibiotics, enzymes, and biogas. It also covers the roles of microbes in sewage treatment, as biocontrol agents, and as biofertilizers.
The document discusses important industrial microorganisms used in biotechnology and their applications. It describes how industrial microbes like bacteria, fungi, yeast, algae and viruses are employed in mass production of chemicals, foods, fuels, enzymes and antibiotics. Specific examples mentioned include using lactobacillus bacteria in yogurt production, streptomyces bacteria for antibiotics like erythromycin, penicillium fungi for penicillin, and yeast for ethanol fermentation. The document outlines properties of useful industrial microbes and how they are categorized based on their metabolic products and the industries they impact.
Microbes are widely used in industry for producing foods, beverages, chemicals, and other products. They play important roles in decomposition, nitrogen fixation, and oxygen production in nature. Common industrial applications of microbes include using yeast to produce alcoholic beverages, lactic acid bacteria to make yogurt and cheese, and fungi and actinomycetes to synthesize antibiotics. Microbes also produce organic acids, amino acids, enzymes, vitamins, and probiotics on an industrial scale.
The document discusses various microorganisms and their applications in industrial processes to produce important products like beverages, organic acids, enzymes, antibiotics, vitamins, dairy products, vaccines, amino acids, and pharmaceutical drugs. It provides details on the microbes used in fermentation processes to manufacture these products on a large scale for commercial use. Specific examples of industrial fermentations include using yeasts to produce beverages, fungi like Aspergillus niger to make citric acid, and bacteria such as Lactobacillus for lactic acid production.
Microorganisms are widely used in industry to produce valuable commercial products through fermentation. Key criteria for industrial microorganisms include producing large amounts of a single product, easy cultivation, genetic stability, and ability to grow rapidly and inexpensively. Common microorganisms and products include:
1) Aspergillus niger produces citric acid. Lactobacillus produces lactic acid. Gluconobacter produces gluconic acid.
2) Vitamins like riboflavin are produced by fungi like Ashbya gossypii. Vitamin B12 is produced by bacteria grown in cobalt.
3) Enzymes such as amylase, produced by Asper
The document discusses the production of several pharmaceutical products through microbial fermentation processes. It describes the fermentation of penicillin using Penicillium chrysogenum, the production of citric acid using Aspergillus niger, and the microbial synthesis of vitamin B12, glutamic acid, and other compounds. Key details provided include microbial strains used, fermentation media composition, culture methods, and process parameters.
The basic food law is intended to assure consumers that foods are pure and wholesome, safe to eat, and produced under sanitary conditions. Generally, food law prohibits importation and distribution of food products that are adulterated, or have labels that are false or misleading in any context.
Soil and water conditions. ...
Keep an eye on the forecast for heavy rainfall events. ...
Calibrate, inspect, and maintain manure application equipment. ...
Separation distances for land application. ...
Irrigation of manure sources. ...
Savvy stockpiling and dry manure management.
Based on the mode of action, the major food preservation techniques can be categorized as: (1) slowing down or inhibiting chemical deterioration and microbial growth, (2) directly inactivating bacteria, yeasts, molds, or enzymes, and (3) avoiding recontamination before and after processing.
Food processing waste is derived from the processing of biological materials and is, in the main, biodegradable. Biowaste is defined in the landfill directive as 'waste capable of undergoing anaerobic or aerobic decomposition such as food and garden waste, and paper and cardboard
Water plays a key role in food processing and has various scientific uses. It acts as a solvent, carrier, and lubricant in processes like washing, leaching, extraction, and cooling. Proper management and treatment of water is important for food safety and quality in processing plants.
The environmental damage of food production from conventional agriculture is not limited to deforestation and pollutants associated with crop growth. Harvesting the crop represents a significant amount of nutrients, water, and energy being taken from the land.
The basis for sanitation is the removal of soils from the manufacturing environment. There are many benefits to this process. From a food safety standpoint, there is the removal of pathogenic organisms, prevention of the formation of biofilms and removal of potentially harmful chemicals from food contact surfaces.
Food packaging is defined as enclosing food to protect it from tampering or contamination from physical, chemical, and biological sources, with active packaging being the most common packaging system used for preserving food products.
Sugar, salt, nitrites, butylated hydroxy anisol (BHA), butylated hydroxyl toluene (BHT), tert-butylhydroquinone (TBHQ), vinegar, citric acid, and calcium propionate are all chemicals that preserve foods. Salt, sodium nitrite, spices, vinegar, and alcohol have been used to preserve foods for centuries.
Sugaring is a food preservation method similar to pickling. Sugaring is the process of desiccating a food by first dehydrating it, then packing it with pure sugar. This sugar can be crystalline in the form of table or raw sugar, or it can be a high sugar density liquid such as honey, syrup or molasses.
Removing the moisture from food helps prevent bacterial and fungal growth which would ruin stored foods. Smoking is a method of drying that also imparts flavor to the food (usually meat items), and smoke helps keep bacteria-carrying-insects away during the drying process.
Microwave penetrates inside the food materials resulting in entire internal cooking of whole volume of food rapidly and uniformly reducing the processing time and energy. This fast heat transfer in turn results in preservation of nutrients, vitamins contents, flavor, sensory characteristics, and color of food
Food irradiation (the application of ionizing radiation to food) is a technology that improves the safety and extends the shelf life of foods by reducing or eliminating microorganisms and insects. Like pasteurizing milk and canning fruits and vegetables, irradiation can make food safer for the consumer
Food irradiation is the process of exposing food and food packaging to ionizing radiation, such as from gamma rays, x-rays, or electron beams. Wikipedia
Low dose (up to 1 kGy): Inhibit sprouting (potatoes, onions, yams, garlic)
Lowering the temperature of food so that microbes and enzymes are inactivated.
Moisture is changed to ice and microbes become inactive without water.
Packaging food maintains the colour, flavour and texture.
Fast freezing (-25ºC) helps maintain nutritive value and texture of food.
Chilling is an important activity in food processing. Foods are chilled to extend shelf life by reducing biochemical reactions and microbial activity. Temperature control is essential in order to prevent spoilage and food safety concerns during storage.1
Drying is a mass transfer process consisting of the removal of water or another solvent by evaporation from a solid, semi-solid or liquid. This process is often used as a final production step before selling or packaging products.
Food drying is a method of food preservation in which food is dried (dehydrated or desiccated). Drying inhibits the growth of bacteria, yeasts, and mold through the removal of water.
Dehydration has been used widely for this purpose since ancient times; the earliest known practice is 12,000 B.C. by inhabitants of the modern Middle East and Asia regions. Drying is a simple method for preserving food.
Dried foods make great healthy and tasty snacks. They are good for lunches, travel, backpacking, hiking, and camping plus many other activities. Most types of foods can be dried. Drying is an ancient method of food preservation.
Most foods will not support the growth of bacteria if their water activity is less than 0.85, because at this water activity there is not enough water available for the bacteria to grow.
However, yeasts can grow at water activities as low as 0.70, while some molds will grow even at water activities as low as 0.60!
Foods with water activities in this range usually have preservatives added to prevent the growth of yeasts and molds.
Acidic foods with a pH less than 4.6, such as tomato sauce, retard the growth of microorganisms. Thus an acidic food with a water activity less than 0.85 is relatively shelf stable, especially if it is stored in the refrigerator.
In this case, low pH, water activity and temperature combine to provide good insurance against the growth of harmful pathogens.
Sorption is a physical and chemical process by which one substance becomes attached to another.
Sorption includes both adsorption & absorption
e.g., liquids being absorbed by a solid or gases being absorbed by a liquid, cotton dipped in ink.
Sorption the process in which one substance takes up or holds another; adsorption or absorption
Sorption is a process in which a solute moves from a fluid to a particulate solid.
The food sorption isotherm describes the thermodynamic relationship between water activity and the equilibrium of the moisture content of a food product at constant temperature and pressure. ...
The typical shape of an isotherm reflects the way in which the water binds the system.
Water plays many very important roles in food. It affects texture (dry and brittle versus moist and soft), enables the activity of enzymes and chemical reactions to occur (acts as a solvent), supports the growth of microorganisms, makes it possible for large molecules like polysaccharides and proteins to move about and interact, and conducts heat within food.
Many foods such as meat, poultry, seafood, fruits and vegetables are composed of 75% and more water, so water is the most abundant component in many fresh foods. Other foods such as dairy products, and fresh baked goods also contain high levels of water (about 35% or more). Foods that are high in moisture are at
risk of contamination from the growth of microorganisms such as bacteria, yeast and mold, while dry foods like pasta generally have long shelf lives.
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- These are slides of the talk given at IEEE International Conference on Software Testing Verification and Validation Workshop, ICSTW 2022.
1. By
Assist.Prof
Dr. Berciyal Golda. P
VICAS
Attributes required in industrial microbes to be usedas efficient
cloning and expression hosts
2. Introduction
Microbial technology refers to the use of microbes
to obtain a product or service of economic value. It is
also called as Fermentation.
The revolutionary exploitation of microbial genetic
discoveries in the 1970s, 1980s and 1990s depended
heavily upon the solid structure of industrial
microbiology, described above.
Our modern understanding of
the fermentation process comes from the work of the
French chemist Louis Pasteur.
3. The major microbial hosts for production of recombinant proteins are
E. coli, B. subtilis, S. cerevisiae, Pichia pastoris, Hansenula
polymorpha and Aspergillus niger.
The use of recombinant microorganisms provided the techniques and
experience necessary for the successful application of higher
organisms, such as mammalian and insect cell culture, and transgenic
animals and plants as hosts for the production of glycosylated
recombinant proteins.
4.
5. 13
The Production of Commercial Products by
Recombinants Microorganisms
■ Molecular biotechnology can be used to enhance the
production of many commercially important compounds
e.g. Vitamins
Amino acids
Antibiotics
■ We will be investigating the use of recombinant
organisms to improve or enhance the production of :
Restriction enzymes
Ascorbic acid
Microbial synthesis of the dye indigo
Production of xanthan gum
6. Therapeutic Agents
Before the advent of molecular biotechnology most human
proteins were available in only small (limited) quantities.
Today hundreds of genes for human proteins have been cloned,
sequenced, expressed in the host cells and are being tested as
therapeutic agents (drugs) in humans.
7. Industrial Microorganisms & Products
Properties of useful industrial microorganism:
Produces spores or can be easily inoculated
Grows rapidly on a large scale in
inexpensive medium
Produces desired product quickly
Should not be pathogenic
Amenable to genetic manipulation
10. ⦁ Microbes especially yeast have been used from time
immemorial for the production of beverages like
wine, beer, whiskey, brandy or rum.
⦁ For this purpose, the
yeast Saccharomyces cerevisiae is used for
fermenting malted cereals and fruit
juices to produce ethanol.
Wine producing bacteria Fungai
Cyberlindnera mrakii
Pichia fermentans
Acetobacter cerevisiae
Lactobacillus bucheri
actobacillus hilgardii
Lactobacillus kunkeei
11.
12. ⦁ Antibiotics produced by microbes are regarded was
one of the most significant discoveries of the
twentieth century and have made major contributions
towards the welfare of human society.
⦁ Many antibiotics are produced by microorganisms,
predominantly by Actinomycetes in the
genus Streptomycin (e.g. Tetracycline, Streptomycin,
Actinomycin D) and by filamentous
fungi (e.g. Penicillin, Cephalosporin)
13. Antibiotic Producer organism Activity Site or mode of action
Penicillin Penicillium chrysogenum Gram-positive bacteria Wall synthesis
Cephalosporin Cephalosporium acremonium Broad spectrum Wall synthesis
Griseofulvin Penicillium griseofulvum Dermatophytic fungi Microtubules
Bacitracin Bacillus subtilis Gram-positive bacteria Wall synthesis
Polymyxin B Bacillus polymyxa Gram-negative bacteria Cell membrane
Amphotericin B Streptomyces nodosus Fungi Cell membrane
Erythromycin Streptomyces erythreus Gram-positive bacteria Protein synthesis
Neomycin Streptomyces fradiae Broad spectrum Protein synthesis
Streptomycin Streptomyces griseus Gram-negative bacteria Protein synthesis
Tetracycline Streptomyces rimosus Broad spectrum Protein synthesis
Vancomycin Streptomyces orientalis Gram-positive bacteria Protein synthesis
Gentamicin
Rifamycin
Micromonospora purpurea
Streptomyces mediterranei
Broad spectrum
Tuberculosis
Protein synthesis
Protein synthesis
15. ⦁ Microbes are also used for the commercial and industrial
production of certain organic acids.
These compounds can be produced directly
from glucose (e.g. gluconic acid) or formed as end
products from pyruvate or ethanol.
Examples of acids producing microorganisms are
Aspergillus Niger (a fungus) of Citric
acid, Acetobacter acute (a bacterium) of Acetic Acid,
Lactobacillus (a bacterium) of lactic acid and many
others.
18. ⦁ Amino acids such as Lysine and Glutamic acid are
used in the food industry as nutritional supplements in
bread products and as flavor enhancing compounds
such as Monosodium Glutamate (MSG).
Amino acids are generally synthesized as primary
metabolites by microbes.
However, when the rate and amount of synthesis of
some amino acids exceed the
cell’s need for protein synthesis, then cell excrete
them into the surrounding medium.
21. ⦁ Many microbes synthesize and excrete large quantities
of enzymesinto the surrounding medium.
⦁ Using this feature of these tiny organisms, many
enzymes have been produced commercially.
⦁ These include Amylase, Cellulase, Protease, Lipase,
Pectinase, Streptokinase, and many others.
⦁ Enzymes are extensively used in food
processing and preservation, washing
powders, leather industry, paper industry and in
scientific research.
25. ⦁ Vitamins are some organic compounds which are
capable of performing many life-sustaining
functions inside our body.
These compounds cannot be
synthesized by humans, and therefore they have to be
supplied in small amounts in the diet.
⦁ Microbes are capable of synthesizing the vitamins and
hence they can be successfully used for the commercial
production of many of the vitamins
e.g. thiamine, riboflavin, pyridoxine, folic acid,
pantothenic acid, biotin, vitamin b12, ascorbic
acid, beta-carotene (pro-vitaminA), ergosterol
(provitamin D)
26. ⦁ Vitamin B12 produced by Propionibacterium
freudenreichii, Pseudomonas denitrificans, Bacillus
megaterium and Streptomyces olivaceus, p.shermanii and etc
⦁ Riboflavin produced by Ashbya gossypii and
Eremothecium ashbyii,clostridum buytilcum,mycocandida
riboflavina,candida flareri and etc,.
⦁ β– Carotene is a pro vitamin produced by Blakeslea
trispora, Phycomyces blakesleeanus and Choanephora
cucurbitarum.
⦁ Blakeslea trispora commenly used for high yield production.
27.
28. ⦁ Organic solvents such as ethanol, acetone, butanol,
and glycerolare some very important chemicals that
are widely used in petrochemical industries.
⦁ These chemicals can be commercially produced by
using microbes and low-cost raw materials
⦁ (e.g. wood, cellulose, starch).
⦁ Yeast (Saccharomyces cerevisiae) is used for
commercial production of ethanol.
31. ⦁ Single Cell Protein (SCP) can serve as an alternate
source of energy when a larger portion of the world is
suffering from hunger and malnutrition.
⦁ Single cell proteins are microbial cells that are rich
in protein content and can be used as protein
supplements for humans and animals.
⦁ Microbes like Spirulina can be grown easily on materials
like waste water from potato processing plants
(containing starch), straw, molasses, animal
manure, and even sewage, to produce large quantities and
can serve as food rich in protein, minerals, fats,
carbohydrate, and vitamins.
32.
33.
34. ⦁ These are a very important group of chemicals, which
are used as anti-inflammatory drugs, and as
hormones such as estrogens and progesterone, which
are used in oral contraceptives.
⦁ Steroids are widely distributed in animals, plants, and
fungi like yeasts.
⦁ But, producing steroids from animal sources or
chemically synthesizing them is difficult, but
microorganisms can synthesize steroids from sterols
or from related, easily obtained compounds.
⦁ Mostly mycobacterium sp are usd frequently.
36. ⦁ Many pharmaceutical drugs are also produced
by microbes
e.g. Cyclosporin A, that is used as an
immunosuppressive agent in organ-transplant
patients, is produced by the fungus Trichoderma
polysporum.
⦁ immunosuppressant cyclosporin A.Statins produced
by the yeast Monascus purpureus have been
commercialized as blood-cholesterol lowering agents.
It acts by competitively inhibiting the enzyme
responsible for the synthesis of cholesterol.
37.
38. Microbes are used in dairy industry to make dairy
product such as curd, yogurt,cheese,kefir , kumies,bread
and various types of milk product.
Saccharomyces cerevisiae,
Streptococcus sp,
penicillium roqueforti,
p.camemberti,
streptococcus thermophilus,
lactobacillus bulgaricus,
Lactobacillus sp,candida sp.
39. ❖Bacterial Transformation
•
• The ability of bacteria to
take in DNA from their
surrounding environment
Bacteria must be made
competent to take up
DNA
Microorganisms as Tools
40. Yeast are Important Too!
❖Single celled eukaryote
❖Kingdom: Fungi
❖Over 1.5 million species
❖Source of antibiotics, blood cholesterol lowering
drugs
❖Able to do post translational modifications
❖Grow anaerobic or aerobic
❖Examples: Pichia pastoris (grows to a higher
density than most laboratory strains), has a no.
of strong promoters, can be used in batch
processes
43. TYPES OF VECTORS
1) Cloning Vectors
Propagation or cloning of DNA insert inside a suitable host
cells. Examples: Plasmids, Phage or Virus
Obtaining millions of copies.
Uses :- Genomic library.
Preparing probes.
Genetic Engineering Experiments.
Selection of cloning vector depends on :-
(a)Objective of cloning experiment
(b)Ease of working.
(c) Knowledge existing about the vector.
(d)Suitability.
(e) Reliability.
44. 2) Expression Vectors
Express the DNA insert producing specific protein.
They have prokaryotic promoter.
Ribosome binding site.
Origin of replication.
Antibiotic resistance gene.
Expression vectors with strong promoters.
Inducible Expression Vectors.
Eukaryotic expression vectors.
45. VECTORS
• Plasmid
• Bacteriophages
• Cosmid
• Yeast Cloning Vectors
• Ti & Ri Plasmids
TARGET HOST CELL
Bacteria, Streptomyces Bacteria
Bacteria
Yeasts
Transformation of cloned
gene in higher plants.
46. AGENTS USED AS VECTORS
PLASMIDS
BACTERIOPHAGES
COSMID
ARTIFICIAL CHROMOSOME VECTORS
In 1973, Cohen described first successful
construction of recombinant vector.
Plasmid PSC101 - Ecoli
47. PLASMID
Extra chromosomal DNA molecules.
Self replicating.
Double stranded.
Short sequence of DNA.
Circular DNA molecules.
Found in prokaryotes.
CHARACTERISTICS
a. Minimum amount of DNA.
b. Two suitable markers for identification .
c. Single restriction site.
d. More restriction enzyme.
e. Size range 1kg – 200kg.
f. Relaxed replication control.
g. Restriction endonuclease enzyme.
48. THREE TYPES OF PLASMID
1. Fertility plasmids:- can perform conjugation.
2. Resistance plasmids:- contain genes that build a
resistance against antibiotics or poisons.
3. Col plasmids:- contain genes that code for proteins
that can kill bacteria.
49. BACTERIOPHAGE VECTORS
Cloning Vectors.
It infects bacteria.
Commonly used Ecoli phages :-
λ phage
M13 Phage
Lambda phage vector
Genome size is 48,502 bp.
High transformation efficiency.
1000 times more efficient than the plasmid vector.
Origin of replication.
Genome linear in head.
Single- stranded protruding cohesive ends of 12 bases.
Cos site – site of cleavage of phage DNA.
50. COSMIDS
Combine parts of the lambda chromosome with parts of
plasmids.
Contain the cos sites of λ and plasmid origin of
replication.
Behave both as plasmids and as phages.
Cosmids can carry up to 50 kb of inserted DNA.
Structure of Cosmid
Origin of replication (ori).
Restriction sites for cleavage and insertion of foreign
DNA.
Selectable marker from plasmid.
A cos site - a sequence yield cohesive end (12 bases).
Ampicillin resistance gene (amp).
51. ARTIFICIAL CHROMOSOME
Linear or Circular.
1 0r 2 copies per cell.
Different types –
Bacterial Artificial Chromosome(BAC)
Yeast Artificial Chromosome (YAC)
P1 derived artificial chromosome (PAC)
Mammalian Artificial Chromosome (MAC)
Human Artificial Chromosome. (HAC)
YAC – Cloning in yeast
BAC & PAC – Bacteria
MAC & HAC – Mammalian & Human cells.
52. YEAST ARTIFICIAL CHROMOSOME
Linear Plasmid Vector.
Clone large DNA segment ( 100 – 1400kb).
Occurring two forms:-
Circular – grows in bacteria. Linear –
multiplies in yeast cells.
pYAC3 - first YAC developed.
It contains :-
ARS sequence – replication
CEN4 sequence – centromeric function TRP1 &
URA3 – 2 selectable markers
Use – mapping complex eukaryotic chromosome .
53. 53
YEAST ARTIFICIAL CHROMOSOMES
•A yeast artificial chromosome (YAC) is a vector used
to clone DNA fragments larger than 100 kb and up to 3000
kb
•YACs are useful for the physical mapping of complex
genomes and cloning of large genes
•A YAC is an artificially constructed chromosome and
contains the telomeric, centromeric, and replication origin
sequences named autonomous replicating sequence
(ARS) needed for replication in yeast cells
54. Why To Use Yeast Vectors?
• Yeast are eukaryotes, contain complex internal
cell structures
• Post-translational modifications
• easy to manipulate as E. coli
• absence of pyrogenic toxins
• cell growth is faster, easier
• less expensive than other eukaryotic cells
• Higher expression levels
• a well-defined genetic system
55. • highly versatile DNA transformation system
• yeast-specific origin of replication (ORI) and a means of
selection in yeast cells, in addition to the bacterial ORI and
antibiotic selection markers
• All contain unique target sites for a number of restriction
endonucleases
• can all replicate in E. coli, often at high copy number
• The four most widely used markers are His3, Leu2, Trp1, and
Ura3.
56. Introducing DNA into fungi
Use of spheroplasts (i.e. wall-less cells) and was
first developed for S. cerevisiae (Hinnen et al., 1978)
Step 1: cell wall is removed enzymically & resulting
spheroplasts are fused with ethylene glycol in the
presence of DNA and CaCl2
Step 2: spheroplasts are then allowed to generate new cell
walls in a stabilizing medium containing 3% agar
Step 3: Electroporation provides a simpler & more convenient
Step 4: DNA can also be introduced into yeasts & filamentous
fungi by conjugation
59. Yeast Integrating plasmids (YIp):
• lack an ORI and must be integrated directly
into the host chromosome via homologous
recombination for efficient multiplication
60. Yeast Episomal plasmids (YEp):
• Beggs (1978)
• recombining an E. coli cloning vector with the naturally
occurring yeast 2 μm plasmid
• 6.3 kb in size
• High copy number of 50–100 per cell
61.
62. Yeast Replicating plasmids (YRp):
• contain anAutonomously Replicating Sequence (ARS)
derived from the yeast chromosome.
• can replicate independently of the yeast chromosome
• unstable and may be lost during budding
63. Yeast centromere plasmids
• carry an ars, most of the recombinants were unstable in yeast
• plasmid-borne centromere sequences have the same distinctive
chromatin structure that occurs in the centromere region of yeast
chromosomes (Bloom & Carbon 1982)
• Three characteristics
• Mitotically stable in the absence of selective pressure
• Segregate during meiosis in a Mendelian manner
• found at low copy number in the host cell
64. Yeast artificial chromosomes
• All autonomous plasmid vectors described above are
maintained in yeast as circular DNA molecules, even the YCp
vectors, which possess yeast centromeres
• Thus, none of these vectors resembles the normal yeast
chromosomes, which have a linear structure
65. • DNAinsert size- 500 kbp
• The ends of all yeast chromosomes, like those of all
other linear eukaryotic chromosomes, have unique
structures that are called telomeres
66. Genes for YAC selection in yeast
• URA3, a gene involved in uracil biosynthesis
• TRP1, a gene involved in tryptophan biosynthesis
• Auxotrophic method of selection
Bacterial replication origin & a bacterial selectable marker
• To propagate the YAC vector in bacterial cells, prior to insertion of
genomic
• DNA, YAC vectors usually contain the ColE1 ori and the ampicillin
• resistance gene for growth and analysis in E. coli.
67.
68. • Three main species of yeast
Saccharomyces cerevisiae
Pichia pastoris
Schizosaccharomyces pombe
69. Saccharomyces cerevisiae
• Baker’s yeast
• Single-celled eukaryote
• Grows rapidly (a doubling time of approximately 90 min)
• Simple, defined media
• Many, but not all, of the post-translation modifications
70. • Strong constitutive promoters
Promoters of phosphoglycerate kinase (PGK), glyceraldehyde-
3-phosphate dehydrogenase (GPD) and alcohol
dehydrogenase (ADH1)
• Suffer similar problems as constitutive E. coli expression
systems
• Inducible production
The GAL System
The CUP1 System
71. The GAL System
• Galactose is converted to glucose-6-phosphate by enzymes of
Leloir pathway
• Leloir pathway structural genes (GAL genes) are expressed at
a high level (0.5–1% of total cellular Mrna), galactose as sole
carbon source
• GAL genes promoter- sites for the transcriptional activator
Gal4p
72. Yeast autonomously replicating sequence (ARS1)
• Struhl et al. (1979)
• carry sequences that enable E. coli vectors to replicate in yeast cells
• sequences are known as ars autonomously replicating sequences
• An ars is quite different from a centromere
• ars acts as an origin of replication , Centromer is involved in chromosome
segregation
Yeast telomeres (TEL)
• Telomeres are the specific sequences (5-TGTGGGTGTGGTG-3), present at ends of
chromosomes in multiple copies, necessary for replication & chromosome
maintenance.
73. • Glucose as carbon source - less Gal4p
• Raffinose as carbon source - Gal4p is produced, binds to GAL
structure gene promoters, but a repressor, Gal80p, inhibits its
activity
• Gal80p binds to Gal4p, mask its activation domain
• Unable to recruit the transcriptional machinery
• Galactose- inhibitory effect of Gal80p
74.
75. The CUP1 System
• Copper ions (Cu2+ and Cu+) are essential, toxic at high levels
• S. cerevisiae, copper homeostasis - uptake, distribution and
detoxification mechanisms
• At high concentrations, detoxification is mediated by a copper ion
sensing metalloregulatory transcription factor-Ace1p
• Upon interaction with copper,Ace1p binds DNAupstream of the
CUP1 gene, encodes a metallothionein protein & induces its
transcription
76. • Expression vectors harbouring CUP1 promoter- induce target
gene expression in a copper-dependent fashion
• Can be grown on rich carbon sources, such as glucose, to high
cell density & protein production is initiated by the addition of
copper sulphate (0.5 mM final concentration)
• Drawback-
presence of copper ions in yeast growth media, and indeed in
water supplies
77. BACTERIAL ARTIFICIAL CHROMOSOME
BAC Vector – PBAC108L.
Cloning of large regions of eukaryotic genome.
Origin of replication from bacterium Ecoli F -factor.
BAC vectors are pBACe3.6, pBeloBAC11.
Used in analysis of genomes.
Host for BAC is mutant strain.
Streptomyces isolates have yielded the majority of human, animal,
and agricultural antibiotics, as well as a number of fundamental
chemotherapy medicines. Streptomyces is the largest antibiotic-
producing genus of actinobacteria, producing chemotherapy,
antibacterial, antifungal, antiparasitic drugs
and immunosuppressants.Streptomyces isolates are typically
initiated with the aerial hyphal formation from the mycelium
78. Why To Use Streptomyces Vectors?
One of the most effective strategies to improve the
production of a particular protein or secondary metabolite
in Streptomyces is to over-express the gene, the synthesis gene
cluster, or the positive regulatory gene for this gene/gene cluster.
The most interesting property of Streptomyces is the
ability to produce bioactive secondary metabolites such as
antifungals, antivirals, antitumoral, anti-hypertensives, and mainly
antibiotics and immunosuppressives.
79. What is the contribution of Streptomyces to the pharmaceutical
industry?
In the last 80 years, Streptomyces has made a
massive contribution to the field of medicine, not only through
antibacterial antibiotics, but also through antifungal,
antiparasitic and anticancer compounds.
What is the function of Streptomyces?
Streptomyces is the largest antibiotic-producing genus,
producing antibacterial, antifungal, and antiparasitic drugs, and
also a wide range of other bioactive compounds, such as
immunosuppressants
80. What antibiotic does Streptomyces griseus produce?
Griseus is of both historical and ecological relevance. This
species produces streptomycin, a broad‐spectrum
aminoglycoside antibiotic, that inhibits translation and was the
first clinically deployed antibiotic from Streptomyces to be
discovered.
What does Streptomyces cause?
Streptomyces spp., usually saprophytic to humans, can
cause local cutaneous fistulized nodules known as
actinomycetoma or mycetoma. Severe invasive infections have
seldom been reported, but most cases reported have occurred in
immunocompromised patients
81. How many types of Streptomyces are there?
500 species
Streptomyces, genus of filamentous bacteria of the family
Streptomycetaceae (order Actinomycetales) that includes more
than 500 species occurring in soil and water.
How do Streptomyces grow?
Streptomyces life cycle
This is vegetative mycelia which grows into the media
following germ tube formation after spore germination. ... The
final stage of development sees a transition from aerial hyphae to
highly hydrophobic spore chains, often characterised by the
production of a spore pigment
82. Which genus of actinomycetes is most commonly used in
antibiotic production?
Streptomyces
Today, 80% of the antibiotics are sourced from
the genus Streptomyces, actinomycetes being the
most important
Is Streptomyces good or bad?
The great importance given to Streptomyces is partly
because these are among the most numerous and most versatile
soil microorganisms, given their large metabolite production rate
and their biotransformation processes, their capability of
degrading lignocellulose and chitin, and their fundamental role
in biological
83.
84. Members of the genus Streptomyces are the source for
numerous antibacterial pharmaceutical agents; among the
most important of these are:
Chloramphenicol (from S. venezuelae)
Daptomycin (from S. roseosporus)
Fosfomycin (from S. fradiae)
Lincomycin (from S. lincolnensis)
Neomycin (from S. fradiae)
Nourseothricin.
85. How many antibiotics are produced by the genus Streptomyces?
The model estimated the total number of antimicrobial
compounds that this genus is capable of producing to be of the
order of a 100,000 - a tiny fraction of which has been unearthed so
far.
Why are most antibiotics from Streptomyces?
The production of most antibiotics is species specific, and
these secondary metabolites are important so
the Streptomyces spp. can compete with other microorganisms
that may come in contact, or even within the same genus.
Which is the first antibiotic isolated from species Streptomyces?
Actinomycin
Actinomycin was the first antibiotic
isolated from Streptomyces in 1940, followed by streptomycin
three years later
86. Anticancer medicines
Doxorubicin intercalating DNA. Streptomyces, yielded the
medicines doxorubicin (Doxil), daunorubicin (DaunoXome),
and streptozotocin (Zanosar). Doxorubicin is the precursor
to valrubicin (Valstar), myocet, and pirarubicin. Daunorubicin is
the precursor to idarubicin (Idamycin), epirubicin (Ellence),
and zorubicin.
Streptomyces is the original source
of dactinomycin (Cosmegen), bleomycin (Blenoxane), pingyangm
ycin (Bleomycin A5), mitomycin
C (Mutamycin), rebeccamycin, staurosporine (precursor
to stauprimide and midostaurin), neothramycin, aclarubicin,
tomaymycin, sibiromycin, and mazethramycin.
87. Derivatives of Streptomycetes isolate migrastatin,
including isomigrastatin, dorrigocin A & B, and the synthetic
derivative macroketone, are being researched for anticancer
activity.
88. Antibiotics
Most clinical antibiotics were found during the "golden age of antibiotics"
(1940s–1960s). Actinomycin was the first antibiotic isolated from Streptomyces in
1940, followed by streptomycin three years later.
Antibiotics from Streptomyces isolates (including various aminoglycosides)
would go on to comprise over two-thirds of all marketed antibiotics.
Streptomyces-derived antibiotics include:
Chloramphenicol (Streptomyces venezuelae)
Daptomycin (Streptomyces roseosporus)
Fosfomycin (Streptomyces fradiae)
Lincomycin (Streptomyces lincolnensis)
Neomycin (Streptomyces fradiae)
Platensimycin (Streptomyces platensis)
Puromycin (Streptomyces alboniger)
Streptomycin (Streptomyces griseus)
Tetracycline (Streptomyces rimosus and Streptomyces aureofaciens)
89.
90. ⚫ Streptomyces species -source of medically useful
compounds
⚫ Many biochemical mysteries need tobe elucidated
⚫ Search for novel secondary metabolites
and their biosyntheticgene clusters
⚫ Relevant genomic databases on Streptomyces species constructed.
⚫ Systems biological approaches useful in the discoveryandengineering studiesof
Streptomyces species
91. • Factors affecting antibiotic production:
1. Medium Composition:
• Carbon source
• Nitrogen source
• Inorganic phosphates
• Inorganic salts
• Trace metals
• Precursors
• Inhibitors
• Inducers
2. Fermentation Conditions:
• pH
• Temperature
• Oxygen
• How can determine the target of inhibitor molecule which may inhibit one of
the biological pathways?
There are many different pathwaies can be applied such as reporter essay by using •
the reporter strains