This document discusses immobilized cell technology and its applications in the beer, wine, and dairy industries. It begins with an introduction to immobilization, which involves imprisoning cells or enzymes in a support or matrix. This allows cells to be reused as they are separated from products. The document then discusses specific applications of immobilized cell technology in wine production, beer production, and dairy industry production. It outlines various support materials and immobilization techniques used for each industry.
Cell disruption is the process of breaking open cell walls to extract intracellular fluid and components without damaging them. The goal is an effective disruption while keeping products active. Methods include mechanical techniques like bead beating, blending, and homogenization which use physical force. Non-mechanical techniques involve freeze-thawing, osmotic shock, chemicals, enzymes, or electricity to disrupt cell walls and membranes in different ways. The optimal method depends on cell type and desired outcome.
This document discusses different methods of immobilizing enzymes and cells, including gel entrapment, encapsulation, adsorption, and containment behind barriers. Gel entrapment involves trapping cells in a polymeric network formed by gelling or cross-linking agents. Encapsulation forms a continuous membrane around cells. Adsorption adheres bacterial cells to a support matrix through various forces. Immobilized cells and enzymes have applications in wastewater treatment and biodiesel production.
This document provides information on different types of fermentation processes including aerobic fermentation, anaerobic fermentation, batch fermentation, fed-batch fermentation, semi-continuous fermentation, and continuous fermentation. It discusses the key differences between aerobic and anaerobic fermentation, and describes the typical phases of batch fermentation. Information is also given on characteristics and applications of fed-batch, semi-continuous, and continuous fermentation.
This document discusses the development of inoculum for industrial fermentation processes. It defines inoculum as a mixture of cultured microbes and the media they are growing in. The key steps in inoculum development are preparing a suitable growth media, maintaining optimal pH and nutrient levels, and conducting growth in stepwise increasing volumes. Examples of common inoculum media compositions are provided for vitamin and bacterial insecticide production processes. Developing high quality inoculum is important for efficiently adapting cultures to fermentation conditions.
This document discusses different methods for immobilizing whole cells, including perfusion bioreactors and biofilm formation. Perfusion bioreactors culture cells continuously over long periods by feeding fresh media and removing waste, while various separation methods like hollow fiber membranes or centrifuges keep cells in the bioreactor. Perfusion offers advantages like improved product quality, smaller reactor size, and lower costs compared to traditional fed-batch systems. The document also covers immobilizing cells through entrapment in polymers, attachment to surfaces, or passive biofilm formation on supports.
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.
Upstream bioprocessing involves steps like isolation and selection of microorganisms, media preparation, inoculation and incubation. Downstream bioprocessing involves steps like product harvesting, extraction, purification, quality control and packaging. Major upstream steps are formulation of fermentation medium, sterilization, inoculum preparation and fermentation. Downstream steps include cell disruption, solid-liquid separation, concentration, purification, formulation and quality monitoring. The overall process aims to isolate the desired product from fermentation broth in pure form through various unit operations.
1. A fluidized bed bioreactor is similar to a bubble column bioreactor but with an expanded top section.
2. In a fluidized bed bioreactor, microorganisms or cell cultures are grown while suspended in a liquid medium by the upward flow of gas or air bubbles from below.
3. This helps maintain the organisms in suspension and provides conditions for optimum growth, such as gas-liquid contact and mixing, while minimizing shear stress on the cells.
Cell disruption is the process of breaking open cell walls to extract intracellular fluid and components without damaging them. The goal is an effective disruption while keeping products active. Methods include mechanical techniques like bead beating, blending, and homogenization which use physical force. Non-mechanical techniques involve freeze-thawing, osmotic shock, chemicals, enzymes, or electricity to disrupt cell walls and membranes in different ways. The optimal method depends on cell type and desired outcome.
This document discusses different methods of immobilizing enzymes and cells, including gel entrapment, encapsulation, adsorption, and containment behind barriers. Gel entrapment involves trapping cells in a polymeric network formed by gelling or cross-linking agents. Encapsulation forms a continuous membrane around cells. Adsorption adheres bacterial cells to a support matrix through various forces. Immobilized cells and enzymes have applications in wastewater treatment and biodiesel production.
This document provides information on different types of fermentation processes including aerobic fermentation, anaerobic fermentation, batch fermentation, fed-batch fermentation, semi-continuous fermentation, and continuous fermentation. It discusses the key differences between aerobic and anaerobic fermentation, and describes the typical phases of batch fermentation. Information is also given on characteristics and applications of fed-batch, semi-continuous, and continuous fermentation.
This document discusses the development of inoculum for industrial fermentation processes. It defines inoculum as a mixture of cultured microbes and the media they are growing in. The key steps in inoculum development are preparing a suitable growth media, maintaining optimal pH and nutrient levels, and conducting growth in stepwise increasing volumes. Examples of common inoculum media compositions are provided for vitamin and bacterial insecticide production processes. Developing high quality inoculum is important for efficiently adapting cultures to fermentation conditions.
This document discusses different methods for immobilizing whole cells, including perfusion bioreactors and biofilm formation. Perfusion bioreactors culture cells continuously over long periods by feeding fresh media and removing waste, while various separation methods like hollow fiber membranes or centrifuges keep cells in the bioreactor. Perfusion offers advantages like improved product quality, smaller reactor size, and lower costs compared to traditional fed-batch systems. The document also covers immobilizing cells through entrapment in polymers, attachment to surfaces, or passive biofilm formation on supports.
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.
Upstream bioprocessing involves steps like isolation and selection of microorganisms, media preparation, inoculation and incubation. Downstream bioprocessing involves steps like product harvesting, extraction, purification, quality control and packaging. Major upstream steps are formulation of fermentation medium, sterilization, inoculum preparation and fermentation. Downstream steps include cell disruption, solid-liquid separation, concentration, purification, formulation and quality monitoring. The overall process aims to isolate the desired product from fermentation broth in pure form through various unit operations.
1. A fluidized bed bioreactor is similar to a bubble column bioreactor but with an expanded top section.
2. In a fluidized bed bioreactor, microorganisms or cell cultures are grown while suspended in a liquid medium by the upward flow of gas or air bubbles from below.
3. This helps maintain the organisms in suspension and provides conditions for optimum growth, such as gas-liquid contact and mixing, while minimizing shear stress on the cells.
Measurement of mass transfer coefficient (k la) Ashok Shinde
The document discusses measurement of the volumetric mass transfer coefficient (KLa), which indicates the rate of oxygen transfer in a bioreactor. It describes various methods to determine KLa values, including chemical and physical techniques like the sodium sulphite oxidation method. The document also covers factors that affect KLa, and how KLa values are used to scale bioreactors from laboratory to production scale.
This document provides an overview of fermentation technology and downstream processing. It defines fermentation as the production of a product by microorganism mass culture. It describes the basic stages of batch fermentation including lag, log, stationary and death phases. It then outlines the main steps in downstream processing including removal of insolubles, product isolation, purification, polishing and packaging. Specific unit operations used at each stage like centrifugation, filtration, chromatography are also explained. The document emphasizes that the level of downstream processing depends on the target product and its end use.
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
The document discusses the continuous production of ethanol from fermentation and purification in a single vessel. The key steps described are:
1) Milling and liquefaction of raw materials like corn to release sugars;
2) Fermentation of sugars to ethanol using yeast;
3) Distillation and rectification to separate and purify ethanol from byproducts; and
4) Possible further processing like evaporation of stillage to produce animal feed.
The overall process aims to efficiently produce ethanol fuel from renewable sources in a single continuous vessel.
This document discusses the key components required for microbial growth and fermentation, including carbon, nitrogen, minerals, vitamins and oxygen. It outlines the goals of optimizing fermentation media to maximize product yield while minimizing undesirable byproducts. Finally, it examines various carbon sources, nitrogen sources, minerals, trace elements and antifoaming agents used in fermentation media formulation.
Immobilization of cells involves encapsulating cells in polymers to prevent division while maintaining viability. This allows increasing cell density in bioreactors for higher metabolite production. Methods include entrapment in gels, on fiber mats, or behind semi-permeable membranes. Alginate is commonly used for gel entrapment. Immobilized systems offer advantages like prolonged biomass use, higher product yields, and genetic stability. They also simplify downstream processing when products are extracellular. Various bioreactor designs are used depending on the immobilization method. Applications show enhanced production of metabolites like serpentine, anthraquinones, and capsaicin using immobilized plant cells.
Microbial Kinetics in Batch Culture
Culture system containing a limited amount of nutrient, which is inoculated with the microorganism. Cells grow until some component is exhausted or until the environment changes so as to inhibit growth. Biomass concentration defined in terms of cell dry weight measurements (g/l) or total cell number (cells/ml).
Lineweaver-Burke Equation.....We remember the Monod Equation
Invert…
The equation now has the form of a straight line with intercept.
Y = MX + C
By plotting as a function of
You get a straight line, where the slope is , and the y–axis intercept is .
Product Yield Coefficient
Maintenance:
Cells use energy and raw materials for two functions, production of new cells and the maintenance of existing cells. In general, consumption of materials for maintenance is small w.r.t. the amount of materials used in the synthesis of new biomass.
Generally it is assumed that the use of materials for maintenance is proportional to the amount of cells present.
Downstream processing refers to the stages involved after fermentation or bioconversion, including separation, purification, and packaging of the product. The key stages are removal of insolubles through filtration, centrifugation or flocculation, product isolation using techniques like liquid-liquid extraction or adsorption, product purification using chromatography or crystallization, and product polishing which prepares the product for packaging and storage. Downstream processing aims to recover and purify the target product from the fermentation or reaction broth.
Batch, fedbatch and continuous fermentationDhanya K C
The document discusses different types of fermentation processes including batch, fed-batch, and continuous fermentation. It explains the key characteristics of each type such as whether the system is open or closed, and how substrates and cells are added or removed. The stages of microbial cell growth including lag phase, exponential phase, stationary phase, and death phase are also summarized for batch fermentation.
Steps involved in fermentation products producing a viable product output.various steps and process were explained in them. A semester syllabus of undergraduate microbiology student in his/her semester -5 in paper -6 . I think this might be helpful to you and have a good response after reading this .thank you.
The document discusses inoculum development and production media for industrial fermentation. It defines inoculum as a culture of microbes used to inoculate production-scale fermentations. Successful fermentations require developing inoculum to an active, healthy state in appropriate density. The document outlines factors that affect fermentation and discusses various media components like carbon sources, nitrogen sources, and trace elements. It also covers inoculum development methods for bacterial and mycelial cultures, preservation techniques, examples of media used for specific inocula, and criteria for a good inoculum.
This document discusses the airlift fermenter. It notes that fermenters must provide a controlled environment for microorganism or cell growth to produce desired products. An airlift fermenter circulates liquid using the density difference between the riser and downcomer columns caused by sparged air or gas. The main type discussed is the concentric draft tube airlift fermenter, which has an internal riser tube that introduces gas to lift liquid up the riser and down the surrounding downcomer tube. Tower loop and ICI deep shaft airlift fermenters are also mentioned. Airlift fermenters provide mixing without mechanical agitation and have high oxygen transfer rates, making them well-suited
PHB production by bacteria and its applicationsಶಂತನು ಕೆ. ಗೌಡ
Polyhydroxybutyrates (PHBs) are biodegradable polymers produced by some bacteria when excess carbon is available. Bacteria accumulate PHBs intracellularly as carbon and energy reserves. PHB is the most common type and was first discovered in 1925. It has properties suitable for applications like bioplastics, medical implants, and packaging. Research is optimizing production methods like varying carbon sources, nutrients, and growing conditions to improve PHB yields from bacteria. Genetic engineering and mutation studies also aim to develop higher yielding bacterial strains for more economical commercial production of PHB bioplastics.
The document discusses fermentors and bioreactors. It describes how fermentors are closed vessels used for large-scale fermentation processes to produce products like antibiotics, amino acids, and organic acids. The document outlines the key components of fermentors, including a water jacket, stirring paddles, and inputs and outputs for nutrients, products, and steam. It also discusses upstream processing like medium preparation and sterilization, inoculation, and the different types of fermentation systems like batch, continuous, and fed-batch culture. Downstream processing steps like product extraction, purification, and formulation are also summarized.
This document discusses the development of inocula for yeast processes. It begins by defining inoculum as a mixture of culture microbes and the media they are growing in. It then outlines the key constituents of inoculum media, including a carbon and nitrogen source, minerals, and factors to control pH and avoid foaming. The document describes the step-wise process used to develop inocula, transferring contents between vessels of increasing size. It provides specifics on inocula development for brewing and baker's yeast processes, noting they involve multiple aerobic stages and transfers to reduce contamination risks.
Crystallization and whole broth processing are important industrial techniques. Crystallization involves forming solid crystals from a solution, melt, or vapor and is widely used in pharmaceutical and chemical purification. It allows isolation of products with high purity at low cost. Whole broth processing recovers metabolites directly from unfiltered fermentation broth using methods like ion exchange resins, dialysis, expanded-bed adsorption, or resin absorption to minimize inhibitory effects during fermentation. Common equipment for crystallization includes tank crystallizers and forced circulation crystallizers.
This document provides an introduction to bioprocess engineering. It discusses the objectives of bioprocess engineering which include biomass, enzyme, metabolite, and recombinant protein production. The stages of bioprocess development include upstream and downstream processes. Important applications of bioprocess engineering are in agriculture, food production, pharmaceuticals, and environmental remediation. When manufacturing products at large scale, bioprocess engineering principles are important to consider.
The document discusses various strategies for recovery and purification of bio-products from fermentation broth. It describes key unit operations like solid-liquid separation techniques like filtration, centrifugation and flocculation to remove cells and debris. Further purification techniques involve precipitation, solvent extraction, ultrafiltration to concentrate and purify the product. Final processing includes crystallization, drying techniques like lyophilization and spray drying to package the purified product. Cell disruption methods including homogenization, bead mills and ultrasonication are also summarized to release intracellular components.
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.
This document discusses various fermentation techniques used in industrial bioprocesses. It begins by defining fermentation and describing fermentation techniques. There are several types of fermentations described - batch, continuous, fed-batch, anaerobic, aerobic, surface, submerged, and solid-state fermentations. Each type is briefly explained highlighting its key characteristics and industrial applications. Important fermentation products like ethanol, glycerol, lactic acid are also listed. The document concludes by stating that traditional fermentations will remain important in food production and future research should identify risks and benefits of specific indigenous fermented products.
Measurement of mass transfer coefficient (k la) Ashok Shinde
The document discusses measurement of the volumetric mass transfer coefficient (KLa), which indicates the rate of oxygen transfer in a bioreactor. It describes various methods to determine KLa values, including chemical and physical techniques like the sodium sulphite oxidation method. The document also covers factors that affect KLa, and how KLa values are used to scale bioreactors from laboratory to production scale.
This document provides an overview of fermentation technology and downstream processing. It defines fermentation as the production of a product by microorganism mass culture. It describes the basic stages of batch fermentation including lag, log, stationary and death phases. It then outlines the main steps in downstream processing including removal of insolubles, product isolation, purification, polishing and packaging. Specific unit operations used at each stage like centrifugation, filtration, chromatography are also explained. The document emphasizes that the level of downstream processing depends on the target product and its end use.
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
The document discusses the continuous production of ethanol from fermentation and purification in a single vessel. The key steps described are:
1) Milling and liquefaction of raw materials like corn to release sugars;
2) Fermentation of sugars to ethanol using yeast;
3) Distillation and rectification to separate and purify ethanol from byproducts; and
4) Possible further processing like evaporation of stillage to produce animal feed.
The overall process aims to efficiently produce ethanol fuel from renewable sources in a single continuous vessel.
This document discusses the key components required for microbial growth and fermentation, including carbon, nitrogen, minerals, vitamins and oxygen. It outlines the goals of optimizing fermentation media to maximize product yield while minimizing undesirable byproducts. Finally, it examines various carbon sources, nitrogen sources, minerals, trace elements and antifoaming agents used in fermentation media formulation.
Immobilization of cells involves encapsulating cells in polymers to prevent division while maintaining viability. This allows increasing cell density in bioreactors for higher metabolite production. Methods include entrapment in gels, on fiber mats, or behind semi-permeable membranes. Alginate is commonly used for gel entrapment. Immobilized systems offer advantages like prolonged biomass use, higher product yields, and genetic stability. They also simplify downstream processing when products are extracellular. Various bioreactor designs are used depending on the immobilization method. Applications show enhanced production of metabolites like serpentine, anthraquinones, and capsaicin using immobilized plant cells.
Microbial Kinetics in Batch Culture
Culture system containing a limited amount of nutrient, which is inoculated with the microorganism. Cells grow until some component is exhausted or until the environment changes so as to inhibit growth. Biomass concentration defined in terms of cell dry weight measurements (g/l) or total cell number (cells/ml).
Lineweaver-Burke Equation.....We remember the Monod Equation
Invert…
The equation now has the form of a straight line with intercept.
Y = MX + C
By plotting as a function of
You get a straight line, where the slope is , and the y–axis intercept is .
Product Yield Coefficient
Maintenance:
Cells use energy and raw materials for two functions, production of new cells and the maintenance of existing cells. In general, consumption of materials for maintenance is small w.r.t. the amount of materials used in the synthesis of new biomass.
Generally it is assumed that the use of materials for maintenance is proportional to the amount of cells present.
Downstream processing refers to the stages involved after fermentation or bioconversion, including separation, purification, and packaging of the product. The key stages are removal of insolubles through filtration, centrifugation or flocculation, product isolation using techniques like liquid-liquid extraction or adsorption, product purification using chromatography or crystallization, and product polishing which prepares the product for packaging and storage. Downstream processing aims to recover and purify the target product from the fermentation or reaction broth.
Batch, fedbatch and continuous fermentationDhanya K C
The document discusses different types of fermentation processes including batch, fed-batch, and continuous fermentation. It explains the key characteristics of each type such as whether the system is open or closed, and how substrates and cells are added or removed. The stages of microbial cell growth including lag phase, exponential phase, stationary phase, and death phase are also summarized for batch fermentation.
Steps involved in fermentation products producing a viable product output.various steps and process were explained in them. A semester syllabus of undergraduate microbiology student in his/her semester -5 in paper -6 . I think this might be helpful to you and have a good response after reading this .thank you.
The document discusses inoculum development and production media for industrial fermentation. It defines inoculum as a culture of microbes used to inoculate production-scale fermentations. Successful fermentations require developing inoculum to an active, healthy state in appropriate density. The document outlines factors that affect fermentation and discusses various media components like carbon sources, nitrogen sources, and trace elements. It also covers inoculum development methods for bacterial and mycelial cultures, preservation techniques, examples of media used for specific inocula, and criteria for a good inoculum.
This document discusses the airlift fermenter. It notes that fermenters must provide a controlled environment for microorganism or cell growth to produce desired products. An airlift fermenter circulates liquid using the density difference between the riser and downcomer columns caused by sparged air or gas. The main type discussed is the concentric draft tube airlift fermenter, which has an internal riser tube that introduces gas to lift liquid up the riser and down the surrounding downcomer tube. Tower loop and ICI deep shaft airlift fermenters are also mentioned. Airlift fermenters provide mixing without mechanical agitation and have high oxygen transfer rates, making them well-suited
PHB production by bacteria and its applicationsಶಂತನು ಕೆ. ಗೌಡ
Polyhydroxybutyrates (PHBs) are biodegradable polymers produced by some bacteria when excess carbon is available. Bacteria accumulate PHBs intracellularly as carbon and energy reserves. PHB is the most common type and was first discovered in 1925. It has properties suitable for applications like bioplastics, medical implants, and packaging. Research is optimizing production methods like varying carbon sources, nutrients, and growing conditions to improve PHB yields from bacteria. Genetic engineering and mutation studies also aim to develop higher yielding bacterial strains for more economical commercial production of PHB bioplastics.
The document discusses fermentors and bioreactors. It describes how fermentors are closed vessels used for large-scale fermentation processes to produce products like antibiotics, amino acids, and organic acids. The document outlines the key components of fermentors, including a water jacket, stirring paddles, and inputs and outputs for nutrients, products, and steam. It also discusses upstream processing like medium preparation and sterilization, inoculation, and the different types of fermentation systems like batch, continuous, and fed-batch culture. Downstream processing steps like product extraction, purification, and formulation are also summarized.
This document discusses the development of inocula for yeast processes. It begins by defining inoculum as a mixture of culture microbes and the media they are growing in. It then outlines the key constituents of inoculum media, including a carbon and nitrogen source, minerals, and factors to control pH and avoid foaming. The document describes the step-wise process used to develop inocula, transferring contents between vessels of increasing size. It provides specifics on inocula development for brewing and baker's yeast processes, noting they involve multiple aerobic stages and transfers to reduce contamination risks.
Crystallization and whole broth processing are important industrial techniques. Crystallization involves forming solid crystals from a solution, melt, or vapor and is widely used in pharmaceutical and chemical purification. It allows isolation of products with high purity at low cost. Whole broth processing recovers metabolites directly from unfiltered fermentation broth using methods like ion exchange resins, dialysis, expanded-bed adsorption, or resin absorption to minimize inhibitory effects during fermentation. Common equipment for crystallization includes tank crystallizers and forced circulation crystallizers.
This document provides an introduction to bioprocess engineering. It discusses the objectives of bioprocess engineering which include biomass, enzyme, metabolite, and recombinant protein production. The stages of bioprocess development include upstream and downstream processes. Important applications of bioprocess engineering are in agriculture, food production, pharmaceuticals, and environmental remediation. When manufacturing products at large scale, bioprocess engineering principles are important to consider.
The document discusses various strategies for recovery and purification of bio-products from fermentation broth. It describes key unit operations like solid-liquid separation techniques like filtration, centrifugation and flocculation to remove cells and debris. Further purification techniques involve precipitation, solvent extraction, ultrafiltration to concentrate and purify the product. Final processing includes crystallization, drying techniques like lyophilization and spray drying to package the purified product. Cell disruption methods including homogenization, bead mills and ultrasonication are also summarized to release intracellular components.
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.
This document discusses various fermentation techniques used in industrial bioprocesses. It begins by defining fermentation and describing fermentation techniques. There are several types of fermentations described - batch, continuous, fed-batch, anaerobic, aerobic, surface, submerged, and solid-state fermentations. Each type is briefly explained highlighting its key characteristics and industrial applications. Important fermentation products like ethanol, glycerol, lactic acid are also listed. The document concludes by stating that traditional fermentations will remain important in food production and future research should identify risks and benefits of specific indigenous fermented products.
Immobilization is "the imprisonment of an enzyme in a distinct phase that allows exchange with, but is separated from the bulk phase in which the substrate, effector or inhibitor molecules are dispersed and monitored"
Plants are natural sources of valuable secondary metabolites used in pharmaceuticals, agrochemicals, the food industry, etc.
There is an increasing demand to obtain these metabolites through more productive plant tissue applications and cell culture methods.
Plant cells can be immobilized through various methods like surface attachment, entrapment in porous matrices, containment behind barriers, and self-aggregation. This allows maintaining high cell densities to increase productivity of secondary metabolites. Immobilization provides advantages like easier product separation, continuous processing, and protecting cells from shear forces. However, limitations include additional costs, complexity in understanding plant cell pathways, and potential loss of biosynthetic capacity. Applications of immobilized plant cells include production of high-value compounds, biotransformations, and synthetic seed technology.
The document discusses various methods for immobilizing microbial cells and enzymes, including carrier binding techniques like adsorption, covalent binding, cross-linking and entrapment. It describes common support materials like natural polymers, synthetic polymers and inorganic materials used for immobilization. The advantages of immobilization include recyclability, stability and potential applications in industries like food, biomedical and biodiesel production. Yeast cell immobilization using calcium alginate entrapment is provided as an example.
The document discusses the key stages in an industrial bioprocess, including strain selection, laboratory and pilot scale development, and industrial scale-up. Ideal strains are genetically stable, efficiently produce the target product, utilize a variety of low-cost substrates, and are safe. Strain selection involves screening natural environments, purchasing from culture collections, genetic engineering, and inducing mutations. Proper isolation, preservation, and storage of industrial microbial strains is important for bioprocess development.
Enzyme definition, Enzyme immobilization introduction , Enzyme immobilization definition, Explanation about support/ matrix, Examples about immobilized enzymes and their product, Advantages of immobilization, Applications of immobilization, Methods of immobilization in different categories like Adsorption method, Covalent bonding method, Entrapment method, Co polymerization /Cross linking method, Encapsulation method, Applications of immobilized enzymes, Diagrammatic explanation about methods of immobilization.
The document discusses methods for the preparation of probiotic feed, including the selection of microbial strains, fermentation processes, and drying techniques. Key points include:
- Probiotic microbes are selected based on their ability to survive the gastrointestinal environment and manufacturing/storage processes while maintaining viability.
- Fermentation can be done using batch, fed-batch, or continuous methods at optimal temperatures and pH for microbial growth.
- Drying methods like freeze drying and spray drying are used to produce probiotic powders, with freeze drying better maintaining cell viability but being more costly.
This document discusses perfusion culture systems. It begins by defining perfusion culture as a system where waste medium is continuously removed from the culture and replaced with fresh medium while retaining viable cells. It then discusses advantages of perfusion culture like high cell density, productivity and flexibility. It also covers cell retention methods in perfusion bioreactors like alternating tangential flow filtration and centrifugation. The document concludes by noting that perfusion culture can offer benefits like improved efficiency but requires consideration of validation and regulatory issues.
Fermentation is a process that uses microorganisms to produce food, pharmaceuticals, and alcoholic beverages on an industrial scale. There are three main types of fermentation processes: batch, fed-batch, and continuous. Batch fermentation involves adding all nutrients at once and allowing the microbes to grow until maximum product concentration is reached. Fed-batch fermentation involves regularly adding fresh media without removing culture. Continuous fermentation continuously removes and replenishes culture media and products to maintain steady conditions. Fermenters come in various sizes and designs including surface, submerged, stirred tank, airlift, and bubble column fermenters. Proper media formulation providing carbon, nitrogen, minerals and other nutrients is essential for microbial growth
Fermentation in medicinal biotechnologySumitKhandai
This document discusses fermentation in medicinal biotechnology. It describes fermentation as a process used to manufacture various medically important products through microorganisms or mammalian cells in a controlled environment. It outlines different types of fermentation processes including batch, continuous, and fed-batch fermentation. It also discusses various parts of bioreactors used for industrial fermentation like agitation systems, oxygen delivery systems, and controls for temperature and pH. Finally, it summarizes different types of bioreactors used in fermentation including stirred tank, airlift, bubble column, fluidized bed, packed bed, photobioreactor, and membrane bioreactors.
ENZYME IMMOBILIZATION BP 605T BIOTECH.pptxSumant Saini
Immobilization involves attaching enzymes to a support matrix. This allows the enzyme to be reused by retaining it within a bioreactor. Common immobilization methods include adsorption, covalent bonding, entrapment, and encapsulation. The technique used depends on factors like strength of attachment and effect on enzyme activity. Immobilized enzymes have various applications in industries like pharmaceuticals, food production, and waste water treatment due to advantages like enhanced stability and easier product separation.
The document discusses various methods for isolating and preserving microorganisms in pure culture. To isolate microbes, common methods include streak plating, pour plating, and serial dilution. Maintaining pure cultures long-term involves subculturing to fresh media periodically or preservation through lyophilization, low-temperature storage, or overlaying cultures with mineral oil. Lyophilization involves freeze-drying microbes under vacuum to remove water and stop metabolic activity, allowing long-term viability.
The document discusses various aspects of biopharmaceutical production including bioreactors, upstream and downstream processing, and specific production methods. It describes how bioreactors provide an effective environment for cell growth and product expression. Upstream processing involves optimizing cell culture and bioreactor conditions while downstream processing focuses on purification techniques like filtration, chromatography. Specific examples discussed include antibiotic production using bacteria and fungal fermentation as well as plant cell culture for secondary metabolite production.
Biotechnology for solving slime problems in the pulpBABU P
This document discusses how biotechnology can help solve problems with biological slimes (biofilms) in the pulp and paper industry. It describes how biofilms form and become resistant to traditional biocides. It then explains how biosurfactants, enzymes, and biodispersants produced using biotechnology can be used to control slimes. Specifically, it discusses how biosurfactants can emulsify oils and inhibit biofilm formation, enzymes can target extracellular polysaccharides to disrupt biofilms, and biodispersants can enhance biocide penetration and efficacy.
The word Fermentation is derived from Latin word fervere which means to boil.
But the conventional definition of Fermentation is to break down of larger molecules into smaller and simple molecules using microorganisms.
In Biotechnology, Fermentation means any process by which microorganisms are grown in large quantities to produce any type of useful materials.
The document discusses various methods for producing and preserving starter cultures for fermented dairy products. It describes traditional liquid culture production methods and highlights that they are time-consuming and risk contamination. It then outlines several improvements to culture production including concentrated, freeze-dried cultures and cryoprotected frozen cultures that allow for direct inoculation and overcome issues with traditional methods. The document also discusses factors that affect survival of freeze-dried cultures and outlines three main systems for bulk starter culture production: using simple techniques; mechanically protected tanks; and propagation in a phage-inhibitory medium.
This document discusses the medical applications of fermentation technology. It begins with an introduction to fermentation and how microorganisms can be used to produce useful chemicals. It then discusses the types and stages of industrial fermentation processes. Some key applications of fermentation in medicine discussed include the production of insulin, vaccines, interferons, vitamin B12, enzymes, and antibiotics. Modern fermentation allows for mass production of these substances using genetically engineered microorganisms.
Cell immobilization is the process of fixing intact cells onto specific regions in a device or material without losing their biological function. Cells can be immobilized through physical adsorption, encapsulation, entrapment, and self-aggregation.
The document provides information on various food regulatory organizations around the world:
- It discusses key Indian acts and organizations that regulate food including the Food Safety and Standards Act 2006, AGMARK, and FSSAI.
- The Food and Agriculture Organization (FAO) is introduced as the UN agency working to defeat hunger internationally.
- Details are given about the structure and departments of the FAO, as well as its objectives, programs, and achievements.
- The US Food and Drug Administration is summarized, including what products it regulates and its organizational structure with centers focused on specific product areas.
- Other food safety systems discussed include HACCP, which provides a systematic approach to food safety,
This document discusses various ways that whey, a byproduct of cheese production, can be utilized. It is commonly used as animal feed but can also be concentrated, dried into powder, or processed into protein concentrates. Further processing can produce refined lactose, ethanol, organic acids, yeast products, biogas, hydrogen, or electricity through microbial fuel cells. The highest value applications involve separation or fermentation processes to extract high value components from whey for use in foods, pharmaceuticals, or energy production.
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive functioning. Exercise causes chemical changes in the brain that may help protect against mental illness and improve symptoms.
This document discusses microbial flavors produced by microorganisms through metabolic processes. It describes several types of flavor compounds produced, including lactones, alcohols, aldehydes, and methyl ketones. Methods for microbial flavor production are outlined, such as de novo synthesis, biotransformation, and enzymatic methods using microbes like yeast, bacteria, and fungi. Examples of specific flavor compounds produced through these methods and the microbes involved are provided. Advantages of microbial flavor production are noted. The current status and market for microbial flavors is briefly summarized.
Microbial pigments can be produced by microorganisms like bacteria, algae, and fungi through solid substrate or submerged fermentation. These natural pigments are promising alternatives to synthetic colors which are toxic. Various screening methods like Raman spectroscopy and mass spectrometry can be used to identify pigment-producing microbial strains. Microbial pigments have advantages like stability, potential health benefits, and ability to be produced sustainably. Future research focuses on genetic engineering to improve pigment yields and utilizing waste materials as substrates.
This document discusses high fructose corn syrup (HFCS) and maltose syrup. It describes the process for making HFCS, which involves processing corn starch with enzymes like amylases and glucose isomerase to convert glucose into fructose. An alternative method uses inulinase enzymes on inulin to produce 95% fructose yield. Maltose syrup is made through a process of mixing starch with water and enzymes like amylases, then filtering, concentrating, and crystallizing to produce a syrup high in maltose. Both HFCS and maltose syrup are used as sweeteners in food production.
This document discusses glucose syrup and invert sugar syrups. Glucose syrup is made from starch hydrolysis and typically contains 10-43% glucose. It is produced through soaking, gelatinization, hydrolysis, clarification, and evaporation steps. Invert syrup contains equal proportions of glucose and fructose produced through acid or enzymatic hydrolysis of sucrose. Both syrups are used as sweeteners and thickeners in foods like candy, ice cream, and baked goods due to properties like moisture retention and flavor enhancement. They have applications in confectionery, pharmaceuticals, and as flavoring agents.
The document summarizes the production processes of yoghurt and kefir. Yoghurt is made by fermenting milk with bacteria that convert lactose into lactic acid. The milk is standardized, pasteurized, homogenized, cooled, inoculated with cultures, fermented, cooled, flavored, and packaged. Kefir is made by inoculating milk with kefir grains containing bacteria and yeast. The microbes produce a polysaccharide that causes the milk to thicken during fermentation. Kefir grains are separated and reused in the process.
This document provides information on traditional Indian fermented foods. It begins with an introduction on fermentation and the benefits it provides foods. Some common Indian fermented foods are then described, including dosa, idli, and dhokla made from rice and legumes, and dairy products like curd, shrikhand, buttermilk, and yogurt. The microorganisms involved in fermenting these foods are noted. Fermented pickles and vegetables like gundruk and sinki are also discussed. The document concludes with brief descriptions of fermented fish products in India like ngari and hentak.
This document discusses cultured dairy milk and cream. It begins by defining cultured dairy milk as fermented milk produced using lactic acid bacteria. It then describes the composition and types of cultured dairy milk products. The main types discussed are cultured buttermilk, Bulgarian buttermilk, acidophilus milk, kefir, kumiss, cultured sour cream, and yogurt. Manufacturing processes are provided for some products. The document also discusses cultured cream, describing types like creme fraiche, sour cream, and clotted cream. It concludes by noting the advantages and disadvantages of cultured dairy products.
- Cheese is produced from milk through coagulation of the casein protein using rennet or lactic acid bacteria, which results in curds that are further processed (e.g. pressed, aged).
- The basic steps of cheese production are receiving/standardizing milk, adding cultures/rennet, coagulation, cutting/cooking curds, pressing/drying, and aging.
- There are various types of cheese classified by moisture content and aging process, from soft (high moisture) to hard/very hard (low moisture).
This document summarizes information about lactic acid bacteria and fermented foods. It discusses how lactic acid bacteria play an important role in food fermentation and preservation through producing lactic acid. Common fermented foods produced with lactic acid bacteria include sauerkraut, pickles, yogurt, cheese, buttermilk, kimchi, idli and dosa. The document provides details on the microorganisms involved and production processes for several of these fermented foods.
Lactic acid can be produced through fermentation by microorganisms. It has various industrial uses, especially in cosmetics, pharmaceuticals, chemicals, food, and medical industries. Lactic acid fermentation occurs in wooden fermenters of 25-125 klt capacity using organisms like Lactobacillus kept at temperatures between 30-50°C depending on the species. The pH is maintained between 5.5-6.5 through additions of calcium carbonate or hydroxide and the fermentation takes 5-10 days to complete. Purification includes filtration, acidification, washing, evaporation and passing through ion exchange resins to obtain 50-60% pure lactic acid.
Citric acid is a natural organic acid found in plants and animals. It is widely used in food and pharmaceuticals, with global production estimated at 736,000 tons per year. Historically, citric acid was first isolated from lemon juice in 1874. Commercially, it is produced using the fungus Aspergillus niger via submerged, surface, or solid state fermentation processes. Submerged fermentation, accounting for 80% of production, involves growing A. niger in a liquid medium in batch, continuous, or fed-batch systems. Citric acid is then harvested through precipitation, extraction, or adsorption/absorption methods.
Vinegar is produced through the fermentation of ethanol by acetic acid bacteria, yielding acetic acid. There are three main methods of production: (1) the Orleans method using wooden barrels, (2) submerged fermentation in large steel tanks with air bubbling, and (3) the generator method dripping alcohol through wood shavings with air blown through. The production process involves refining raw vinegar to remove bacteria, filtration, pasteurization, bottling, and concentration to increase the acetic acid content.
Wine is produced through a multi-step process involving harvesting grapes, crushing and extracting juice, fermenting the juice into alcohol, aging and blending, and finally bottling. Key steps include crushing grapes to extract juice sugars, inoculating the juice with yeast to convert sugars to alcohol through primary and secondary fermentation, aging and developing flavors, removing solids and particles, and bottling the finished wine. Distilled spirits like whiskey and brandy involve additional distillation of wine.
The document discusses the process of whisky production. It involves malting barley grains, mashing and filtering the grains, boiling wort, fermentation using yeast, double distillation to increase alcohol concentration, aging the distilled alcohol in oak barrels for 3-4 years, and finally blending and bottling the aged whisky. Key steps are malting, fermentation, double pot distillation, aging in oak barrels for several years to develop flavors, and blending to produce the final whisky product.
Vodka is produced through fermenting and distilling grains or vegetables to extract alcohol, which is then purified and diluted with water. The production process involves mashing the raw materials to convert starches to sugars, sterilizing and inoculating the mash with yeast to facilitate fermentation, distilling the resulting liquid to separate alcohol from other compounds, and bottling the diluted alcohol. Modern technologies have automated many steps but the basic process remains the same.
Rum is produced from sugar cane or its byproducts like molasses through a process of fermentation and distillation. Yeast is added to sugar cane juice or molasses which is then fermented for 1-3 weeks through either spontaneous or controlled fermentation. The fermented mash is then distilled, with heavier rums coming from pot stills and lighter rums from column stills. Rum is then aged in barrels like bourbon or cognac barrels to mature, though there is no minimum aging time.
This document summarizes the beer production process, which includes malting, kilning, milling, mashing, boiling the wort, fermentation, finishing, aging, maturation, and carbonation. During malting, barley grains are soaked, germinated, and dried. Milling crushes the grains. Mashing involves mixing the grist with warm water to produce sugars and flavor components. The wort is boiled with hops added for flavor and sterilization. Fermentation converts sugars to alcohol and carbon dioxide using yeast. Finishing involves aging, clarifying, and carbonating the beer.
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...Sérgio Sacani
Context. With a mass exceeding several 104 M⊙ and a rich and dense population of massive stars, supermassive young star clusters
represent the most massive star-forming environment that is dominated by the feedback from massive stars and gravitational interactions
among stars.
Aims. In this paper we present the Extended Westerlund 1 and 2 Open Clusters Survey (EWOCS) project, which aims to investigate
the influence of the starburst environment on the formation of stars and planets, and on the evolution of both low and high mass stars.
The primary targets of this project are Westerlund 1 and 2, the closest supermassive star clusters to the Sun.
Methods. The project is based primarily on recent observations conducted with the Chandra and JWST observatories. Specifically,
the Chandra survey of Westerlund 1 consists of 36 new ACIS-I observations, nearly co-pointed, for a total exposure time of 1 Msec.
Additionally, we included 8 archival Chandra/ACIS-S observations. This paper presents the resulting catalog of X-ray sources within
and around Westerlund 1. Sources were detected by combining various existing methods, and photon extraction and source validation
were carried out using the ACIS-Extract software.
Results. The EWOCS X-ray catalog comprises 5963 validated sources out of the 9420 initially provided to ACIS-Extract, reaching a
photon flux threshold of approximately 2 × 10−8 photons cm−2
s
−1
. The X-ray sources exhibit a highly concentrated spatial distribution,
with 1075 sources located within the central 1 arcmin. We have successfully detected X-ray emissions from 126 out of the 166 known
massive stars of the cluster, and we have collected over 71 000 photons from the magnetar CXO J164710.20-455217.
When I was asked to give a companion lecture in support of ‘The Philosophy of Science’ (https://shorturl.at/4pUXz) I decided not to walk through the detail of the many methodologies in order of use. Instead, I chose to employ a long standing, and ongoing, scientific development as an exemplar. And so, I chose the ever evolving story of Thermodynamics as a scientific investigation at its best.
Conducted over a period of >200 years, Thermodynamics R&D, and application, benefitted from the highest levels of professionalism, collaboration, and technical thoroughness. New layers of application, methodology, and practice were made possible by the progressive advance of technology. In turn, this has seen measurement and modelling accuracy continually improved at a micro and macro level.
Perhaps most importantly, Thermodynamics rapidly became a primary tool in the advance of applied science/engineering/technology, spanning micro-tech, to aerospace and cosmology. I can think of no better a story to illustrate the breadth of scientific methodologies and applications at their best.
(June 12, 2024) Webinar: Development of PET theranostics targeting the molecu...Scintica Instrumentation
Targeting Hsp90 and its pathogen Orthologs with Tethered Inhibitors as a Diagnostic and Therapeutic Strategy for cancer and infectious diseases with Dr. Timothy Haystead.
The cost of acquiring information by natural selectionCarl Bergstrom
This is a short talk that I gave at the Banff International Research Station workshop on Modeling and Theory in Population Biology. The idea is to try to understand how the burden of natural selection relates to the amount of information that selection puts into the genome.
It's based on the first part of this research paper:
The cost of information acquisition by natural selection
Ryan Seamus McGee, Olivia Kosterlitz, Artem Kaznatcheev, Benjamin Kerr, Carl T. Bergstrom
bioRxiv 2022.07.02.498577; doi: https://doi.org/10.1101/2022.07.02.498577
The technology uses reclaimed CO₂ as the dyeing medium in a closed loop process. When pressurized, CO₂ becomes supercritical (SC-CO₂). In this state CO₂ has a very high solvent power, allowing the dye to dissolve easily.
Authoring a personal GPT for your research and practice: How we created the Q...Leonel Morgado
Thematic analysis in qualitative research is a time-consuming and systematic task, typically done using teams. Team members must ground their activities on common understandings of the major concepts underlying the thematic analysis, and define criteria for its development. However, conceptual misunderstandings, equivocations, and lack of adherence to criteria are challenges to the quality and speed of this process. Given the distributed and uncertain nature of this process, we wondered if the tasks in thematic analysis could be supported by readily available artificial intelligence chatbots. Our early efforts point to potential benefits: not just saving time in the coding process but better adherence to criteria and grounding, by increasing triangulation between humans and artificial intelligence. This tutorial will provide a description and demonstration of the process we followed, as two academic researchers, to develop a custom ChatGPT to assist with qualitative coding in the thematic data analysis process of immersive learning accounts in a survey of the academic literature: QUAL-E Immersive Learning Thematic Analysis Helper. In the hands-on time, participants will try out QUAL-E and develop their ideas for their own qualitative coding ChatGPT. Participants that have the paid ChatGPT Plus subscription can create a draft of their assistants. The organizers will provide course materials and slide deck that participants will be able to utilize to continue development of their custom GPT. The paid subscription to ChatGPT Plus is not required to participate in this workshop, just for trying out personal GPTs during it.
The debris of the ‘last major merger’ is dynamically youngSérgio Sacani
The Milky Way’s (MW) inner stellar halo contains an [Fe/H]-rich component with highly eccentric orbits, often referred to as the
‘last major merger.’ Hypotheses for the origin of this component include Gaia-Sausage/Enceladus (GSE), where the progenitor
collided with the MW proto-disc 8–11 Gyr ago, and the Virgo Radial Merger (VRM), where the progenitor collided with the
MW disc within the last 3 Gyr. These two scenarios make different predictions about observable structure in local phase space,
because the morphology of debris depends on how long it has had to phase mix. The recently identified phase-space folds in Gaia
DR3 have positive caustic velocities, making them fundamentally different than the phase-mixed chevrons found in simulations
at late times. Roughly 20 per cent of the stars in the prograde local stellar halo are associated with the observed caustics. Based
on a simple phase-mixing model, the observed number of caustics are consistent with a merger that occurred 1–2 Gyr ago.
We also compare the observed phase-space distribution to FIRE-2 Latte simulations of GSE-like mergers, using a quantitative
measurement of phase mixing (2D causticality). The observed local phase-space distribution best matches the simulated data
1–2 Gyr after collision, and certainly not later than 3 Gyr. This is further evidence that the progenitor of the ‘last major merger’
did not collide with the MW proto-disc at early times, as is thought for the GSE, but instead collided with the MW disc within
the last few Gyr, consistent with the body of work surrounding the VRM.
hematic appreciation test is a psychological assessment tool used to measure an individual's appreciation and understanding of specific themes or topics. This test helps to evaluate an individual's ability to connect different ideas and concepts within a given theme, as well as their overall comprehension and interpretation skills. The results of the test can provide valuable insights into an individual's cognitive abilities, creativity, and critical thinking skills
1. IMMOBILIZED CELL TECHNOLOGY IN
BEER, WINE AND DAIRY INDUSTRY.
Submitted to : DR. RS singh
Submitted by : mumtaj begum
19011022
MSc. hons .biotechnology 2nd year
PUNJABI UNIVERSITY ,
PATIALA .
2. CONTENTS :
• WHAT IS IMMOBILIZATION.
• INTRODUCTION.
• IMMOBILIZED CELL SYSTEM
• TYPES OF SUPPORTS
• WINE PRODUCTION WITH IMMOBILIZED CELL
• BEER PRODUCTION WITH IMMOBILIZED CELL
• DAIRY INDUSTRY PRODUCTION WITH IMMOBILIZED CELL .
• ADVANTAGES
• DISADVANTAGES
3. INTRODUCTION :
• Immobilization is defined as the imprisonment of cell or
enzymes in a particular support or matrix.
• The immobilized whole cell system is an alternative to
enzyme immobilization where the target cell is immobilized.
• The support or matrix allows the exchange of medium
containing substrate or effector or inhibitor molecule.
• Allows cells to be held in place throughout the reaction,
following which they are easily separated from the products
and may be used again.
5. IMMOBILIZED CELL SYSTEM
• Immobilized cell systems Cells can be kept inside bioreactors in
suspension (free cells) or immobilized in various supports.
• There are four main immobilization techniques for yeast cells:
1. Attachment to a surface,
2. Entrapment within a porous matrix
3. Cell aggregation (flocculation)
4. Containment behind barriers
5
6. 1.Attachment to a surface
• The attachment to a surface can be done by natural adsorption,
electrostatic forces or covalent binding, with cross-linking agents.
• The attachment of cells to an organic or inorganic support may
be obtained also by creating chemical bonds (covalent) between
cells and the support using cross-linking agents.
• However, this immobilization procedure is generally incompatible
with cell viability, since the cross-linking agents are highly toxic
for the microbial cells decreasing their activity.
6
7. • As consequence, this method of immobilization is no longer
used for microbial cells but still remains suitable for the
immobilization of enzymes.
• The surface of the immobilization support is important in the
process of adsorption of cells as rough surfaces allows the cell
retention into the support’s cavities.
• This immobilization technique is often used as it is an easy and
natural process that takes place spontaneously.
• In the last years, natural adsorption is the most used technique
for yeast cell immobilization and further applied in wine making.
7
8. 2.Entrapment within a porous matrix
• Entrapment within a porous matrix can be performed by two
approaches:
• a) The cells are introduced in a porous material and, after
growing, their mobility is restricted by the presence of other cells
and by the matrix
b) A solid matrix is synthesized in situ around the cells. The cells
are incorporated in the matrix of a more or less rigid polymer.
• The polymers are synthetic such as polyacrylamide, or can be
made from proteins (gelatin, collagens) and polysaccharides
(cellulose, alginate, agar, and carrageenan).
8
9. 3.Cell Aggregation or Flocculation
• Cell aggregation or flocculation can occur naturally or by using
artificial flocculating agents.
• It is a complex process connected with the expression of
flocculation genes such as FLO1, FLO5, FLO8 and FLO11.
• Yeast flocculation is an attractive method because of its
simplicity and low costs.
• The flocculation depends on various parameters such as pH,
nutrients, dissolved oxygen, medium composition and
fermentation conditions (temperature and agitation) as well as
the age of the cell.
9
10. • In food industry, the main applications of the flocculation
are the alcohol production, some kind of beers and
sparkling wines (secondary fermentation).
• The flocculation is very important for the brewing industry
as it is an effective, environmentally friendly, easy and
without costs method to separate the yeast cells from beer
at the end of the fermentation.
• The flocculation of the yeast is a very important
characteristic also in the traditional making of sparkling
wines.
10
11. 4.Containment Behind a Barrier
• Containment behind a barrier can be achieved by two main
methods:
• Entrapment of the cells in microcapsules and by the use of
microporous membrane filters (hollow fiber) or by cell
immobilization onto an interaction surface of two immiscible
liquids.
• The method based on the entrapment of cells in microcapsule or
encapsulations, consists firstly in entrapping the cells in a
spherical gel and posterior coating with a polymer such as
polyethyleneimine.
• Then, the gel is dissolved but the cells are left in suspension,
contained behind the polymer barrier.
11
12. • The microporous membranes filters are normally made of
polymers, e.g. polyvinylchloride or polypropylene.
• The containment of the cells behind a barrier allows very high
cell concentrations. For this reason, the membranes used should
be freely permeable to nutrients and products released during
the fermentation , as well as mechanically resistant.
• This method of immobilization is normally used when a cell free
product is needed.
• The main disadvantages are related to mass transfer limitations
and the possibility of membrane fouling caused by the cell
growth.
12
13. Types of supports
• For successful industrial application of this technology the
proposed supports must ideally be of food grade
quality,abundant in nature and cost effective.
• These supports are mostly natural organic polysaccharides or
inorganic material abundant in nature. They may be used
without much modification
or after minor treatment to alter their properties(porosity,
surface charges, etc.), others can be commercially synthesized.
13
14. • Examples of supports and techniques proposed in wine-making
in the recent years include the following:
1) Inorganic supports for cell immobilization in wine-making
2) Organic supports for cell immobilization in wine-making
3) Membrane systems for cell immobilization in wine-making
4) Natural supports for cell immobilization in wine-making
14
15. Wine production with immobilized
cells
• Immobilization technology is used in
various fermentation processes.
• Immobilized cells were used for
bioethanol production,cider production,
vinegar and brewing as well as for wine-
making.
• In our days, the induction of alcoholic
fermentation and malolactic fermentation
is done with starter cultures of cells, i.e.
pure culture of cells isolated and
developed for conducting wine
fermentations.
15
16. • Most fermenters used in the winemaking industry are of a
batch type, i.e. separate lots (batches) and are individually
fermented till conclusion of the process.
• Some industries adopted continuous methods, because of its
advantages in controlling the yeast population and activity,
keeping them in their maximum.
17. • The environmental conditions of
continuous fermentations are favorable for
the yeast growth, thus the biomass
concentration is approximately two times
larger than traditional wine-making.
• One of the most important characteristic
of the continuous process is the high
volumetric productivity but, despite of its
potential advantages, it is only profitable
when working all year-round.
• Immobilized cell systems emerged as a
technique that provides also large amounts
of cells but is more economic than the free
cells continuous wine-making.
17
19. INTRODUCTION TO BREWING
• Brewing is one of the oldest biotechnologies with history back
more than 8000 yrs.
• Brewing is the production of beer through steeping a starch
source (commonly cereal grains) in water and then fermenting
with yeast.
• Ingredients are water, fermentable starch source, brewing yeast,
flavoring agent & secondary starch source(adjunct).
• Main steps involved are
• Malting ,Milling, Mashing
• Wort filteration and boiling ,
• Hop addition, Fermentation ,Aging etc.
19
20. CONTD..
• Fermentation is the essential part of the brewing process,
responsible for the formation of most flavor compounds, while
the secondary fermentation provides beer maturation and final
beer sensory properties.
• They are most time consuming steps in the overall beer
production.
20
21. IMMOBILIZED CELL TECHNOLOGY (ICT) IN BEER
INDUSTRY
• ICT has been attracting continual attention in brewing industry
over past 30 yrs.
• Reasons are:-
▶ Faster fermentation rates.
▶ Increased volumetric productivity.
▶ Continuous operation.
• It is well established now in secondary fermentation and alcohol
free & low alcohol beer production.
• Key parameters of this technology are:
Selections of carrier material
Method of immobilisation together with the bioreactor design.
21
22. CONTD..
• ICT processes have been designed for different stages in beer
production.
• 1. Bioflavoring during maturation
• 2 . Main fermentation
• 3 . Fermentation for production of alcohal free beer and low
alcohol beer.
22
23. ADVANTAGES
• Main advantages of using immobilised cells for production of
beer are:
Enhanced fermentation productivity due to higher biomass
densities
Improved cell stability
Easier implementation of continuous operation
Improved operational control and flexibility
Facilitated cell recovery and reuse
Simplified downstream processing.
23
24. BASIC MODES OF CELL IMMOBILIZATION USED
IN BREWING
• Cell immobilisation can be classified into four categories
based on the mechanism of cell localisation and the nature of
support material:
(i) Attachment to the support surface
(ii) Entrapment within a porous matrix
(iii) Containment behind or within a barrier
(iv) Self-aggregation
24
25. ATTACHMENT TO THE SUPPORT SURFACE
• Micro-organisms adsorb spontaneously on a wide variety of
organic and inorganic supports.
• Yeast cells are immobilised by ionic attraction. Adsorption
affinity of yeast cells for glass was increased by pretreating
the cells in aluminium ions.
• DEAE cellulose supports have been commonly used for the
production of alcohol-free beer and maturation of green beer.
25
26. ENTRAPMENT WITHIN A POROUS MATRIX
• It can be performed by two different basic methods:
first is gel entrapment, the porous matrix is synthesised in situ
around the cells to be immobilised.
In the second, cells are allowed to move into the preformed
porous matrix.
• Both methods provide cell protection from the fluid shear and
higher cell densities.
• The concept is that the matrix is porous enough for substrates and
products to traverse.
• Mechanical strength of matrix is important.
26
27. CONTD..
• Polysaccharides (e.g. alginate, chitosan, pectate and
carrageenan), synthetic polymers (e.g. polyvinylalcohol, PVA)
and proteins (gelatine, collagen) can be used under mild
conditions forentrapment with minimal loss of viability.
• Mainly gel entrapmet is used.
• Gels are mostly used in form of spherical beads with diameters
ranging from about 0.3 to 5 mm.
27
28. CONTAINMENT BEHIND OR WITHIN A BARRIER
It includes
Systems with a barrier formed around cells such as microcapsules
ie. microencapsulation
• Too expensive to be used in beer production
Systems with cells contained within a compartment separated by
a preformed membrane such as hollow fibre and flat membrane
modules.
• Polymeric microfiltration or ultrafiltration membranes, ceramic,
silicone or ion exchange membranes are used.
Mass transfer through the membrane is dependent on the pore
size and structure as well as on the hydrophobicity/hydrophilicity
and surface charge.
28
29. SELF-AGGREGATION
• Passive techniques based on sedimentation capabilities
• Based on formation of cell clumps or floccules, which can be
naturally occurring as in the case of flocculent yeast strains, or
induced by addition of flocculating agents.
• Simplest and the least expensive method.
• It has been successfully exploited over almost 40 years by
Dominion Breweries in New Zealand.
• Flocculent yeast cells are separated from the beer in a conical
settler by gravity. Recycled back into the hold-up vessel to
increase the cell density and to achieve better control of the
fermentation rates.
29
30. CARRIER SELECTION AND DESIGN
A list of the various carrier materials, which have been
investigated for application in beer production, is presented.
30
31. REACTOR DESIGN
• ICT bioreactors can be classified into three categories,
depending on the location of immobilised cells:
• 1. STATIONARY PARTICLE REACTOR
• 2. MOVING AND MIXED PARTICLE REACTOR
• 3. MOVING SURFACE REACTOR
For fermented food or beverage production, bioreactors of
category (i) and (ii) are usually employed
• Various modifications and combinations of stirred tank,
packed-bed, fluidised-bed, gas-lift and membrane
reactors were proposed for different phases in beer
production.
31
32. CONTD..
Selection of the appropriate reactor must be based on critical
issues such as:
Choice of cell carrier
Supply and Removal of gases and solutes in the liquid phase
and removal of excess biomass formed.
Reactor sterilisation and sterile transfer of immobilised
biocatalysts.
• Reactors that can be thoroughly sterilised and directly
inoculated with cells or cell-aggregates or reactors packed
with preformed porous carriers are desireable.
• Primary beer fermentation systems are much more sensitive
to contamination than the secondary fermentation systems.
32
33. CONTD..
• Enterobacteria and acetic acid bacteria have been detected in
immobilised yeast bioreactors used for primary fermentation
while lactic acid bacteria were found in reactors used for
secondary fermentations.
Various methods are available to fight development of
contaminants in an ICT bioreactor:
• Sulphite addition (widely used in the wine industry)
• Heat treatment, are also used in suspended and immobilised
cell systems.
• Use of high dilution rates or harsher environmental conditions
(e.g. pH, temperature, salt concentrations, …)
33
35. INTRODUCTION
Lactic acid bacteria (LAB) are widely used in the production
of fermented dairy products such as cheese, yoghurts and
creams because of their technological, nutritional and
eventual health properties.
The production of organic acids and the resulting
acidification is essential for the production, development of
typical flavour and preservation of these products.
The transformation of lactose by lactic cultures improves the
digestibility and various metabolic and enzymatic activities
of LAB lead to the production of volatile substances, which
contribute to flavour, aroma and texture developments in
fermented dairy products.
35
36. Probiotics are defined as microbial cells which transit the
gastrointestinal tract and which, in doing so, benefit the
health of the consumer.
Among these micro-organisms, LAB and especially
Lactobacilli and Bifidobacteria are already used in many
probiotic dairy products.
The dairy industry involves processing of raw milk into
products such as consumer milk, butter, cheese, yogurt,
condensed milk, dried milk (milk powder), and ice cream,
using processes such as chilling, pasteurization, and
homogenization.
Typical by-products include buttermilk, whey, and their
derivatives.
36
37. The dairy industry posses many practical difficulties:
Increased contamination
High down streaming cost
Unstable products
Coupling of biomass and metabolite productions
In order to overcome these drawbacks, Immobilization Cell
Technology (ICT) is applied. Immobilization refers to the “
technique in which a macromolecule or cell is confined
spatially i.e. they are associated with a support material
either in soluble or insoluble form, which limits its free
movement so that it can be retained there and reused in
successive process runs”.
37
38. Immobilization modifies:
1) Physiology of cells.
2) Affects the sensitivity of LAB to salt and penicillin.
ICT can be used:
To produce starters for the dairy industry
Aspects of biomass production in beads
Continuous cell release from beads
Continuous fermentations with filtration cell recycle are
examined.
38
39. Advantages of Cell Immobilization as compared
with Free-Cell (FC) Systems :
1) High cell density and very high volumetric productivity.
2) Reuse of biocatalysts.
3) High process stability (physical and biological) over long
fermentation periods.
4) Retention of plasmid-bearing cells.
5) Improved resistance to contamination.
6) Uncoupling of biomass and metabolite productions.
7) Stimulation of production and secretion of secondary
metabolites.
39
41. The purpose of these techniques is either to retain high
cell concentrations within the bioreactor or to protect
cells from a hostile environment.
For industrial applications in the food industry, the
carrier material must be non-toxic, readily available
and affordable.
It should also lead to high-cell loading and the cells
should have a prolonged viability in the support.
For food applications, the most widely used
immobilization technique is the entrapment of cells
within a food-grade porous polymeric matrix.
41
42. Thermal (κ-carrageenan, gellan, agarose, gelatin) or
ionotropic (alginate, chitosan) gelation of the droplets
are used to produce spherical gel biocatalysts.
These polymers are readily available and widely
accepted for use as additives in the food and
particularly dairy.
Gel entrapment is a relatively simple method resulting
in usually spherical beads with diameters ranging
from 0.3 to 3.0 mm with high biomass concentration.
A careful selection of polymer composition is
necessary to achieve high mechanical stability of gel
biocatalysts during long-term fermentation.
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43. LAB Immobilization By Entrapment
technique
SUPPORT SPECIES MAXIMUM CELL
CONCENTRATION
1) Ca-alginate Lactococcus lactis ssp. 2 x 1011 cfu ml-1
2) Ca-alginate Lactococcus lactis ssp. 3.8 x 1011 cfu ml-1
3) k-carrageenan-LBGb Lactobacillus casei 5.1 x 1011 cfu ml-1
4) k-carrageenan-LBGb Lactococcus lactis ssp. 1.3x1011 cfu g-1
5) Gellan gum Bifidobacterium longum 6.8 x 1010 cfu g-1
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44. Various dairy products
Yogurt production:
• Yogurt posses organoleptic, nutritional and eventually
therapeutic qualities. Continuous yogurt prefermentation of
milk in a stirred tank reactor by entrapped cells in Calcium
alginate was proposed to increase performance.
Cheese manufacturing:
ICT demonstrates
High productivity
Better control of mixed culture with impact in the quality
Economic process
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45. Quarg Cheese
A soft, unripened, cow's milk cheese with the texture
and flavor of sour cream. Quarg comes in two versions-
low-fat and nonfat .
The method of Quarg Cheese through ICT have been
proposed:
Direct Vat inoculation with alginate immobilized freeze
dried starter
Inoculation of milk
Proliferation of milk to pH 5.5 in a bioreactor
containing bead with subsequent fermentation to ph
4.6 carried out by cell released by beads.
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46. Cream fermentation:
Advantages Of ICT:
• Better control of pH of final product
• Can use of a continuous system upto month
Frozen desserts
Yogurts cultures, Bifidobacterium , Lb.
acidophilus are used for this preparation.
ICT is responsible for improving quality of such
products by increasing properties of living cells
in the product.
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47. • The preparation of lactose-hydrolysed milk and whey,
using β -galactosidase.
• This is of great significance in a country like India where
lactose intolerance is quite prevalent.
• Lactose hydrolysis also enhances the sweetness and
solubility of the sugars, and can find future potentials in
preparation of a variety of dairy products.
• Lactose-hydrolysed whey may be used as a component
of whey-based beverages, leavening agents, feed stuffs,
or may be fermented to produce ethanol thus converting
an inexpensive byproduct into a highly nutritious, good
quality food ingredient.
• The first company to commercially hydrolyse lactose in
milk by immobilized lactase was Centrale del Latte of
Milan, Italy, utilizing the Snamprogetti technology. The
process makes use of a neutral lactase from yeast
entrapped in synthetic fibres.
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48. • Specialist Dairy Ingredients, a joint venture between the Milk
Marketing Board of England and Wales and Corning, had set up
an immobilized β -galctosidase plant in North Wales for the
production of lactose-hydrolysed whey.
• The β -galactosidase of fungal origin has been used for this
purpose.
• An immobilized preparation obtained by cross-linking β -
galactosidase in hen egg white (lyophilized dry powder) has
been used for the hydrolysis of lactose.
• A major problem in the large-scale continuous processing of
milk using immobilized enzyme is the microbial contamination
which has necessitated the introduction of intermittent
sanitation steps.
• A co-immobilizate obtained by binding of glucose oxidase on
the microbial cell wall using Con A has been used to minimize
the bacterial contamination.
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49. Control of contaminants
Yeast contamination
Yeast is present due to introduction of unwanted microorganism.
Yeast introduction in bioreactor containing lactobaccilli
immobilized alginate beads, is not able to grow in system because
of rapid substrate turnover.
Bacteriophages contamination:
It is a major concern for dairy industries in lactic fermentations.
Most bacteriophages aren’t destroyed by temperatures used for
milk pasteurization and raw milk will introduce phages in the
plant.
When LAB are immobilized in alginate beads, the presence of
bacteriophages doesn’t significantly affect the acidification rate.
For ICT processes designed for the continuation of milk ,free cells
are not protected from phages. Thus phage control strategies have
to be developed or phage resistant strain are used.
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50. Use of Recycled immobilized cell
• Use of immobilized LAB as affinity matrices for removal of
unwanted compounds in liquid.
• Removal of heavy or radioactive metals from waste streams.
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51. REFERENCES
C P Champagne, C Lacroix, I Sodini-GallotCritical Reviews in
Biotechnology1994; 14(2):109-34
http://www.nisco.ch/download/27.pdf
Cell Immobilization For The dairy Industry by Yann Doleyres
and Christophe Lacroix
www.sciencedirect.com
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