1) The document discusses the use of biosensors for detecting heavy metals using bioluminescent bacterial sensors. It describes how reporter genes like luciferase are induced by heavy metals, causing the bacteria to luminesce.
2) Methods of immobilizing bacteria in calcium alginate beads for use in biosensors are covered. The document also examines how biosensors can be used to test environmental samples and monitor toxicity over time and with reuse.
3) Detection techniques like colorimetric assays are discussed, where changes in color can indicate heavy metal presence without specialized instruments. The document evaluates biosensor performance in detecting various heavy metals in water and soil.
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.
Release of genetically engineered microbes in the enviromentKaayathri Devi
Genetically engineered microbes are being released into the environment. However, this could have unintended consequences that are difficult to foresee. More research is needed to fully understand the risks before widespread release of genetically modified microorganisms.
Plants can be used as bioreactors to produce valuable compounds. Transgenic plants and plant cell cultures can produce large quantities of proteins, vaccines, and other molecules through biochemical reactions using techniques like genetic engineering. Some key advantages of plant bioreactors are that they are cost-effective, can produce high biomass, and allow storage of products for a long time. However, differences in plant and bacterial genetics can impact expression efficiency and safety testing is required.
Biosorption uses inactive microbial biomass to bind and concentrate heavy metals from aqueous solutions, even very dilute ones. It is a promising alternative to traditional chemical precipitation for treating industrial effluents due to its low cost and high metal binding capacity. Biosorption is a metabolically passive process where heavy metals bind to functional groups on the cell surface through mechanisms like ion exchange, complexation, and chelation. Algae, fungi, bacteria, and plants have all been studied for their ability to biosorb and bioremediate heavy metals through various metabolic and non-metabolic pathways.
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.
Biosensors show the potential to complement laboratory-based analytical methods for
environmental applications. Although biosensors for potential environmental-monitoring
applications have been reported for a wide range of environmental pollutants, from a regulatory
perspective the decision to develop a biosensor method for an environmental application should
consider several interrelated issues. These issues are discussed in terms of the needs, policies,
and mechanisms associated with the identification and selection of appropriate monitoring
methods.
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.
Release of genetically engineered microbes in the enviromentKaayathri Devi
Genetically engineered microbes are being released into the environment. However, this could have unintended consequences that are difficult to foresee. More research is needed to fully understand the risks before widespread release of genetically modified microorganisms.
Plants can be used as bioreactors to produce valuable compounds. Transgenic plants and plant cell cultures can produce large quantities of proteins, vaccines, and other molecules through biochemical reactions using techniques like genetic engineering. Some key advantages of plant bioreactors are that they are cost-effective, can produce high biomass, and allow storage of products for a long time. However, differences in plant and bacterial genetics can impact expression efficiency and safety testing is required.
Biosorption uses inactive microbial biomass to bind and concentrate heavy metals from aqueous solutions, even very dilute ones. It is a promising alternative to traditional chemical precipitation for treating industrial effluents due to its low cost and high metal binding capacity. Biosorption is a metabolically passive process where heavy metals bind to functional groups on the cell surface through mechanisms like ion exchange, complexation, and chelation. Algae, fungi, bacteria, and plants have all been studied for their ability to biosorb and bioremediate heavy metals through various metabolic and non-metabolic pathways.
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.
Biosensors show the potential to complement laboratory-based analytical methods for
environmental applications. Although biosensors for potential environmental-monitoring
applications have been reported for a wide range of environmental pollutants, from a regulatory
perspective the decision to develop a biosensor method for an environmental application should
consider several interrelated issues. These issues are discussed in terms of the needs, policies,
and mechanisms associated with the identification and selection of appropriate monitoring
methods.
The document outlines India's biosafety guidelines for research and development activities involving genetically modified organisms (GMOs). It discusses the four biosafety levels established by the Department of Biotechnology (DBT) and the containment facilities recommended for each level. It also categorizes experiments into three categories - Category I experiments which are exempt from approval, Category II experiments which require institutional biosafety committee intimation, and Category III experiments which require prior approval from competent authorities. The document provides examples of experiments that fall under each category and discusses the roles of different committees like IBSC, RCGM, GEAC that oversee and regulate GMO research in India.
•Introduction of bioremediation: Bioremediation refers to the process of using microorganisms to remove the environmental pollutants i.e. toxic wastes found in soil, water, air etc.
•In situ bioremediation:
It involves a direct approach for the microbial
degradation of xenobiotics at the sites of pollution
(soil, ground water).
•Types of in situ bioremediation:
Natural attenuation.
Engineered in situ bioremediation.
- Bioventing, biosparging, bioslurping,
phytoremediation.
•Ex situ bioremediation:
Waste or toxic pollutants can be collected from the polluted sites and bioremediation can be carried out at a designated place or site.
• Types of ex situ bioremediation
Land farming, windrow, biopiles, bioreactors.
•Microorganisms use in bioremediation:
A number of naturally occurring marine microbes
such as Pseudomonas sp. is capable of degrading oil and other hydrocarbons.
•Factors affecting bioremediation:
Nutrient availability, moisture content, pH, temperature, contaminant availability.
•References:
Satyanarayana U. Biotechnology. BOOKS AND ALLIED (P) Ltd.
Sharma P.D. Environmental Microbiology. RASTOGI PUBLICATIONS.
Gupta P.K. Biotechnology and Genomics. RASTOGI PUBLICATIONS.
Dubey R.C. A Textbook of Biotechnology. S Chand And Company Ltd.
Dubey R.C. A Textbook of Microbiology. S Chand And Company Ltd.
Willey/Sherwood/Woolverton. Prescott’s Microbiology. McGRAW-HILL INTERNATIONAL EDITION.
www.sciencedirect.com/bioremediation.
1. Acetone-butanol fermentation is a process that uses Clostridium bacteria to produce acetone and butanol from glucose through anaerobic fermentation.
2. Clostridium acetobutylicum and Clostridium beijerinckii are commonly used species that produce a 3:6:1 ratio of acetone, butanol, and ethanol.
3. Historically this fermentation process was used commercially until the 1970s to produce acetone and butanol, which are used as solvents and in producing synthetic rubbers, lacquers, and explosives.
Transgenic plants with biotic stress resistanceSakeena Asmi
This document discusses transgenic plants with resistance to biotic stress. It begins by defining biotic stress as damage caused by living organisms like bacteria, viruses, fungi and insects. Developing transgenic plants is presented as a way to improve crop yields by making plants resistant to these stresses. Specific examples of transgenic plants containing genes from Bacillus thuringiensis (Bt) that code for insecticidal proteins are described. Bt genes have been introduced into crops like corn, cotton and potatoes to resist pests like rootworms and Colorado potato beetles. While Bt crops have increased yields, there is a risk of pests developing resistance over time if not managed properly.
Hydrocarbon are major constituents of crude oil and petroleum. They can be biodegraded by naturally-occurring microorganisms in freshwater and marine environments under a variety of aerobic and anaerobic conditions. The ability of microorganisms - bacteria, archaea, fungi, or algae - to break down hydrocarbons is the basis for natural and enhanced bioremediation. To promote biodegradation, amendments such as nitrogen and phosphorous fertilizer are often added to stimulate microbial growth and metabolism
Bioremediation uses microorganisms or plants to remove pollutants from the environment. There are two main types - in situ treats pollutants on site, while ex situ removes pollutants to off-site facilities. Examples of in situ techniques include bioventing, biosparging, and in situ biodegradation which supply oxygen and nutrients to stimulate bacteria. Ex situ methods include slurry and aqueous reactors which process contaminated materials in a contained system. Bioremediation can degrade pollutants like copper but has limitations such as environmental constraints and long treatment time.
Bioleaching of iron, copper, gold. uraniumAnuKiruthika
This document summarizes the process of bioleaching, which uses microorganisms to extract metals like copper, gold, iron, and uranium from ores. It discusses how different bacteria are used to oxidize the metal sulfides in ores, making the metals soluble and able to be extracted. The main methods used are heap leaching and in-situ leaching. Bioleaching has advantages of being low-cost and able to process low-grade ores, but is also time-consuming. Specific examples of how bacteria aid in leaching copper, iron, gold, and uranium are also provided.
Biosensors in Environmental MonitoringSindhBiotech
This lecture is presented by our volunteer Bushra Umer, she is from Karachi, Pakistan, and she is covering "Biosensors in Environmental Monitoring"
For video: https://youtu.be/DoO2Aw7bRrk
bioplastics by microorganisms Polyhydroxyalkanoates And PolyhydroxybutyratePramod Pal
This document discusses bioplastics, which are plastics derived from renewable biomass sources such as vegetable oils, cornstarch, and pea starch. It notes that bioplastics are designed to biodegrade and can break down in either aerobic or anaerobic environments depending on how they are manufactured. Common types of bioplastics include polylactic acid (PLA), polyhydroxyalkanoic acids (PHAs), and polyhydroxybutyrate-co-valerate (PHBVs). The document also discusses the synthesis and production of bioplastics like PHAs and PHB by microorganisms, as well as their applications in packaging, catering, gardening, medical products, and sanitary products
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.
This document provides an introduction to various fermentation processes and products commonly found in the Indian market, including probiotics and yogurt. It defines fermentation as the chemical transformation of organic substances by microorganisms like bacteria, molds, or yeasts. The document outlines five categories of fermentation processes and provides examples of probiotic microorganisms like various Lactobacillus and Bifidobacterium species. It also lists the microbial contents of common probiotic supplements like Yakult and ViBact and discusses functional properties of yogurt like aiding lactose digestion and inhibiting harmful bacteria.
This document summarizes microbial degradation of various xenobiotics and pollutants. It discusses how microbes like bacteria, fungi and actinomycetes are able to degrade compounds like hydrocarbons, PAHs, pesticides, dyes and other xenobiotics. The microbes produce enzymes that allow them to use these compounds as carbon and energy sources and breakdown the compounds into simpler molecules like carbon dioxide and water.
This document provides information about bioremediation. It begins with an introduction defining bioremediation as using microorganisms to degrade hazardous chemicals into less toxic forms. It then discusses the types of microorganisms involved, including Pseudomonas genus and Xenobiotics-degrading microorganisms. Several examples of pollutants and degrading microorganisms are given. The mechanisms of bioremediation include aerobic and anaerobic transformations such as respiration, fermentation, and methane fermentation. Factors affecting bioremediation like moisture, nutrients, oxygen levels, pH, temperature, and pollutant characteristics are outlined. Methods of bioremediation include in-situ and ex-situ techniques
This document discusses plant molecular pharming (PMP), which uses plants as bioreactors for producing recombinant pharmaceutical proteins. It covers the definition, history, strategies, host systems, production of antibiotics/enzymes/vaccines in plants, advantages/disadvantages of plant systems, and issues of transgene pollution. Key points include:
- PMP uses whole plants, plant cells or tissues to produce commercially valuable proteins like vaccines via recombinant DNA.
- Early work in 1986 produced human growth hormone in tobacco and sunflower. Commercial production of various proteins in plants has occurred.
- Strategies include transforming host plants, growing biomass, processing/purifying the product of interest.
- Plants,
Transfection is a technique used to insert foreign nucleic acids like DNA or RNA into cells to alter their properties. There are various biological, chemical, and physical methods to accomplish transfection, either transiently or stably. In stable transfection, the foreign DNA integrates into the cellular genome and is passed to daughter cells, while transient transfection only expresses the DNA for a short time without integration. Genetic engineering techniques are used to transfer genes between organisms, like retrovirus-mediated gene transfer where the retrovirus acts as a vector to deliver transgenes into host cells. Embryonic stem cell-mediated gene transfer involves introducing DNA into stem cells that can integrate randomly or through homologous recombination and be passed to offspring. Liposome
Bioprocess development and technology-Introduction,History of bioprocess,Milestones of Bioprocess development,Bioprocess development,Impact on Biotechnology
Control systems are necessary in fermenters to carefully monitor and regulate parameters like temperature, pH, oxygen levels, agitation and foaming. Sensors integrated directly into the fermenter provide real-time readings of these parameters to control systems which can activate mechanisms to precisely adjust the fermentation process as needed through elements like heating/cooling systems, pumps to add acids/bases and valves to control gas flow. Proper monitoring and control of these critical parameters is essential for optimal microbial growth and product formation.
Bioleaching is a process that uses microorganisms like bacteria and fungi to extract metals from ores. It involves microbes transforming metal compounds into soluble forms that can then be recovered. Some key microbes used are Thiobacillus ferrooxidans and Thiobacillus thiooxidans, which produce acids that dissolve metals. Bioleaching is commercially done through methods like slope leaching, heap leaching, and in situ leaching. It provides a cost-effective way to extract low-grade ores and is more environmentally friendly than smelting. However, it is a slower process and requires careful control of temperature, pH, and other environmental factors.
Origin and function of a metal resistance gene isolated from eukaryotic soil ...Antoine Ziller
The document discusses a study that analyzed metal resistance genes isolated from eukaryotic soil metatranscriptomes. It found a family of cysteine-rich proteins (CRPs) that were highly expressed in metal-polluted soils. The objectives were to determine if the CRPs were metallothioneins, characterize their functional role, and identify their taxonomic origin. The study compared characteristics of the CRPs to known metallothioneins and found differences in length, cysteine content, cysteine motif occurrence and organization. Experiments were proposed to express CRP genes in E. coli and conduct in vitro metal chelation assays to help elucidate the CRPs' potential role in metal resistance.
The document describes a novel family of biosensors called Nomad biosensors that can measure G protein-coupled receptor (GPCR) activity in living cells. Nomad biosensors detect changes in intracellular second messenger concentrations caused by GPCR activation. Depending on the second messenger involved in the GPCR pathway, there are three versions of Nomad biosensors that detect cAMP, calcium, or DAG. Nomad biosensors have been validated by expressing them in cells with several GPCRs and measuring changes in cytoplasmic granularity following receptor activation. They provide a sensitive method for high-throughput screening of drug libraries to identify compounds that modulate GPCR activity.
The document outlines India's biosafety guidelines for research and development activities involving genetically modified organisms (GMOs). It discusses the four biosafety levels established by the Department of Biotechnology (DBT) and the containment facilities recommended for each level. It also categorizes experiments into three categories - Category I experiments which are exempt from approval, Category II experiments which require institutional biosafety committee intimation, and Category III experiments which require prior approval from competent authorities. The document provides examples of experiments that fall under each category and discusses the roles of different committees like IBSC, RCGM, GEAC that oversee and regulate GMO research in India.
•Introduction of bioremediation: Bioremediation refers to the process of using microorganisms to remove the environmental pollutants i.e. toxic wastes found in soil, water, air etc.
•In situ bioremediation:
It involves a direct approach for the microbial
degradation of xenobiotics at the sites of pollution
(soil, ground water).
•Types of in situ bioremediation:
Natural attenuation.
Engineered in situ bioremediation.
- Bioventing, biosparging, bioslurping,
phytoremediation.
•Ex situ bioremediation:
Waste or toxic pollutants can be collected from the polluted sites and bioremediation can be carried out at a designated place or site.
• Types of ex situ bioremediation
Land farming, windrow, biopiles, bioreactors.
•Microorganisms use in bioremediation:
A number of naturally occurring marine microbes
such as Pseudomonas sp. is capable of degrading oil and other hydrocarbons.
•Factors affecting bioremediation:
Nutrient availability, moisture content, pH, temperature, contaminant availability.
•References:
Satyanarayana U. Biotechnology. BOOKS AND ALLIED (P) Ltd.
Sharma P.D. Environmental Microbiology. RASTOGI PUBLICATIONS.
Gupta P.K. Biotechnology and Genomics. RASTOGI PUBLICATIONS.
Dubey R.C. A Textbook of Biotechnology. S Chand And Company Ltd.
Dubey R.C. A Textbook of Microbiology. S Chand And Company Ltd.
Willey/Sherwood/Woolverton. Prescott’s Microbiology. McGRAW-HILL INTERNATIONAL EDITION.
www.sciencedirect.com/bioremediation.
1. Acetone-butanol fermentation is a process that uses Clostridium bacteria to produce acetone and butanol from glucose through anaerobic fermentation.
2. Clostridium acetobutylicum and Clostridium beijerinckii are commonly used species that produce a 3:6:1 ratio of acetone, butanol, and ethanol.
3. Historically this fermentation process was used commercially until the 1970s to produce acetone and butanol, which are used as solvents and in producing synthetic rubbers, lacquers, and explosives.
Transgenic plants with biotic stress resistanceSakeena Asmi
This document discusses transgenic plants with resistance to biotic stress. It begins by defining biotic stress as damage caused by living organisms like bacteria, viruses, fungi and insects. Developing transgenic plants is presented as a way to improve crop yields by making plants resistant to these stresses. Specific examples of transgenic plants containing genes from Bacillus thuringiensis (Bt) that code for insecticidal proteins are described. Bt genes have been introduced into crops like corn, cotton and potatoes to resist pests like rootworms and Colorado potato beetles. While Bt crops have increased yields, there is a risk of pests developing resistance over time if not managed properly.
Hydrocarbon are major constituents of crude oil and petroleum. They can be biodegraded by naturally-occurring microorganisms in freshwater and marine environments under a variety of aerobic and anaerobic conditions. The ability of microorganisms - bacteria, archaea, fungi, or algae - to break down hydrocarbons is the basis for natural and enhanced bioremediation. To promote biodegradation, amendments such as nitrogen and phosphorous fertilizer are often added to stimulate microbial growth and metabolism
Bioremediation uses microorganisms or plants to remove pollutants from the environment. There are two main types - in situ treats pollutants on site, while ex situ removes pollutants to off-site facilities. Examples of in situ techniques include bioventing, biosparging, and in situ biodegradation which supply oxygen and nutrients to stimulate bacteria. Ex situ methods include slurry and aqueous reactors which process contaminated materials in a contained system. Bioremediation can degrade pollutants like copper but has limitations such as environmental constraints and long treatment time.
Bioleaching of iron, copper, gold. uraniumAnuKiruthika
This document summarizes the process of bioleaching, which uses microorganisms to extract metals like copper, gold, iron, and uranium from ores. It discusses how different bacteria are used to oxidize the metal sulfides in ores, making the metals soluble and able to be extracted. The main methods used are heap leaching and in-situ leaching. Bioleaching has advantages of being low-cost and able to process low-grade ores, but is also time-consuming. Specific examples of how bacteria aid in leaching copper, iron, gold, and uranium are also provided.
Biosensors in Environmental MonitoringSindhBiotech
This lecture is presented by our volunteer Bushra Umer, she is from Karachi, Pakistan, and she is covering "Biosensors in Environmental Monitoring"
For video: https://youtu.be/DoO2Aw7bRrk
bioplastics by microorganisms Polyhydroxyalkanoates And PolyhydroxybutyratePramod Pal
This document discusses bioplastics, which are plastics derived from renewable biomass sources such as vegetable oils, cornstarch, and pea starch. It notes that bioplastics are designed to biodegrade and can break down in either aerobic or anaerobic environments depending on how they are manufactured. Common types of bioplastics include polylactic acid (PLA), polyhydroxyalkanoic acids (PHAs), and polyhydroxybutyrate-co-valerate (PHBVs). The document also discusses the synthesis and production of bioplastics like PHAs and PHB by microorganisms, as well as their applications in packaging, catering, gardening, medical products, and sanitary products
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.
This document provides an introduction to various fermentation processes and products commonly found in the Indian market, including probiotics and yogurt. It defines fermentation as the chemical transformation of organic substances by microorganisms like bacteria, molds, or yeasts. The document outlines five categories of fermentation processes and provides examples of probiotic microorganisms like various Lactobacillus and Bifidobacterium species. It also lists the microbial contents of common probiotic supplements like Yakult and ViBact and discusses functional properties of yogurt like aiding lactose digestion and inhibiting harmful bacteria.
This document summarizes microbial degradation of various xenobiotics and pollutants. It discusses how microbes like bacteria, fungi and actinomycetes are able to degrade compounds like hydrocarbons, PAHs, pesticides, dyes and other xenobiotics. The microbes produce enzymes that allow them to use these compounds as carbon and energy sources and breakdown the compounds into simpler molecules like carbon dioxide and water.
This document provides information about bioremediation. It begins with an introduction defining bioremediation as using microorganisms to degrade hazardous chemicals into less toxic forms. It then discusses the types of microorganisms involved, including Pseudomonas genus and Xenobiotics-degrading microorganisms. Several examples of pollutants and degrading microorganisms are given. The mechanisms of bioremediation include aerobic and anaerobic transformations such as respiration, fermentation, and methane fermentation. Factors affecting bioremediation like moisture, nutrients, oxygen levels, pH, temperature, and pollutant characteristics are outlined. Methods of bioremediation include in-situ and ex-situ techniques
This document discusses plant molecular pharming (PMP), which uses plants as bioreactors for producing recombinant pharmaceutical proteins. It covers the definition, history, strategies, host systems, production of antibiotics/enzymes/vaccines in plants, advantages/disadvantages of plant systems, and issues of transgene pollution. Key points include:
- PMP uses whole plants, plant cells or tissues to produce commercially valuable proteins like vaccines via recombinant DNA.
- Early work in 1986 produced human growth hormone in tobacco and sunflower. Commercial production of various proteins in plants has occurred.
- Strategies include transforming host plants, growing biomass, processing/purifying the product of interest.
- Plants,
Transfection is a technique used to insert foreign nucleic acids like DNA or RNA into cells to alter their properties. There are various biological, chemical, and physical methods to accomplish transfection, either transiently or stably. In stable transfection, the foreign DNA integrates into the cellular genome and is passed to daughter cells, while transient transfection only expresses the DNA for a short time without integration. Genetic engineering techniques are used to transfer genes between organisms, like retrovirus-mediated gene transfer where the retrovirus acts as a vector to deliver transgenes into host cells. Embryonic stem cell-mediated gene transfer involves introducing DNA into stem cells that can integrate randomly or through homologous recombination and be passed to offspring. Liposome
Bioprocess development and technology-Introduction,History of bioprocess,Milestones of Bioprocess development,Bioprocess development,Impact on Biotechnology
Control systems are necessary in fermenters to carefully monitor and regulate parameters like temperature, pH, oxygen levels, agitation and foaming. Sensors integrated directly into the fermenter provide real-time readings of these parameters to control systems which can activate mechanisms to precisely adjust the fermentation process as needed through elements like heating/cooling systems, pumps to add acids/bases and valves to control gas flow. Proper monitoring and control of these critical parameters is essential for optimal microbial growth and product formation.
Bioleaching is a process that uses microorganisms like bacteria and fungi to extract metals from ores. It involves microbes transforming metal compounds into soluble forms that can then be recovered. Some key microbes used are Thiobacillus ferrooxidans and Thiobacillus thiooxidans, which produce acids that dissolve metals. Bioleaching is commercially done through methods like slope leaching, heap leaching, and in situ leaching. It provides a cost-effective way to extract low-grade ores and is more environmentally friendly than smelting. However, it is a slower process and requires careful control of temperature, pH, and other environmental factors.
Origin and function of a metal resistance gene isolated from eukaryotic soil ...Antoine Ziller
The document discusses a study that analyzed metal resistance genes isolated from eukaryotic soil metatranscriptomes. It found a family of cysteine-rich proteins (CRPs) that were highly expressed in metal-polluted soils. The objectives were to determine if the CRPs were metallothioneins, characterize their functional role, and identify their taxonomic origin. The study compared characteristics of the CRPs to known metallothioneins and found differences in length, cysteine content, cysteine motif occurrence and organization. Experiments were proposed to express CRP genes in E. coli and conduct in vitro metal chelation assays to help elucidate the CRPs' potential role in metal resistance.
The document describes a novel family of biosensors called Nomad biosensors that can measure G protein-coupled receptor (GPCR) activity in living cells. Nomad biosensors detect changes in intracellular second messenger concentrations caused by GPCR activation. Depending on the second messenger involved in the GPCR pathway, there are three versions of Nomad biosensors that detect cAMP, calcium, or DAG. Nomad biosensors have been validated by expressing them in cells with several GPCRs and measuring changes in cytoplasmic granularity following receptor activation. They provide a sensitive method for high-throughput screening of drug libraries to identify compounds that modulate GPCR activity.
Biotechnology is an indigenous wave of innovation. This enhances the quality of the environment by protecting the natural resources. It plays key role for sustainable agriculture.
Mitochondria are organelles found in plant cells that provide energy and regulate important metabolic processes. Plant mitochondrial genomes vary significantly in size but generally encode proteins involved in oxidative phosphorylation as well as rRNAs and tRNAs. These genomes often contain introns, open reading frames, and chloroplast DNA sequences. Mutations in mitochondrial DNA can impact plant development and cause cytoplasmic male sterility. Expression of chimeric mitochondrial genes is associated with some cases of male sterility. Studies examine the role of mitochondrial proteins like uncoupling proteins and alternative oxidases in conferring stress tolerance in plants. The WA352 mitochondrial gene is implicated in cytoplasmic male sterility in rice through interaction with the nuclear-encoded protein COX11.
The document discusses using MALDI-TOF mass spectrometry to identify bacteria through protein biomarkers. MALDI-TOF allows analysis of intact bacterial cells by generating species-specific fingerprints from cell envelope components. These fingerprints can be entered into a database to rapidly identify unknown bacterial isolates.
This document summarizes an experiment to engineer ascorbate peroxidase (APX) activity into cytochrome c peroxidase (CCP) by introducing the APX ascorbate-binding site into CCP. Specifically, the researchers replaced the ascorbate-binding loop and a critical arginine residue in CCP with the corresponding residues from APX to create a mutant called CCP2APX. While wild-type CCP showed no APX activity, CCP2APX was able to catalyze the peroxidation of ascorbate, demonstrating that the engineered ascorbate-binding site could bind ascorbate. Crystal structures of CCP2APX confirmed that the engineered binding site
This document summarizes a study that identified a novel reactive chlorine species (RCS) defense mechanism in the bacterium Azospira suillum. The study found:
1) A sigma factor (SigF) and its anti-sigma factor (NrsF) regulate expression of genes involved in RCS defense, including a methionine-rich periplasmic protein (MrpX) and a methionine sulfoxide reductase (YedY1).
2) MrpX is strongly induced by RCS like hypochlorite and acts to scavenge it through methionine oxidation, while YedY1 regenerates reduced MrpX.
Genomic Analysis Of Nitrogen Fixation In Synechococcus Sp. PCC 7002 and Chlor...Heather Jordan
This document summarizes research being conducted on nitrogen fixation in the cyanobacteria Synechococcus sp. PCC 7002 and the green sulfur bacterium Chlorobium tepidum. For Synechococcus, the researcher aims to knock out the NAR1 gene and observe the phenotypic effects. For C. tepidum, the researcher previously inactivated the nifD gene and aims to create a similar nifD knockout construct. The document provides background information on both organisms, including their optimal growth conditions and genetic transformation methods.
Comparison of three different Bioleaching systems for Li recovery from lepido...Suby Mon Benny
Nature article about Lithium Bioextraction by J. Sedlakova-Kadukova, R. Marcincakova, A. Luptakova, M. Vojtko, M. Fujda and P. Pristas explained in a simple manner.
This document reports on a laboratory project investigating the role of the non-coding RNA rli60 in Listeria monocytogenes. The student constructed an rli60 knockout strain of L. monocytogenes and examined gene expression and metabolism compared to the wild type strain when grown in different media. Quantitative PCR results showed that rli60 is involved in down-regulating the ilv operon, which encodes for branched amino acid biosynthesis. Growth curve and luminescence assays indicated normal bacterial growth and hly virulence gene expression in the rli60 mutant strain. In summary, this project found that the non-coding RNA rli60 regulates metabolic gene expression but does not affect growth or a key virulence factor in L. monocytogenes
The document discusses the role of siderophores in plant pathogen interactions. It provides background on siderophores, their importance in microbial metabolism, types of siderophores produced by different organisms, and their mechanism of iron acquisition. It summarizes several case studies that demonstrate how siderophore-producing bacteria can be used for biocontrol of plant pathogens through competition for iron and activation of plant defense responses. Siderophores are shown to elicit plant defenses, modulate signaling pathways, and promote bacterial growth during infection.
Metabolic Pathways in Higher Plants and their DeterminationDr. Siddhi Upadhyay
a) Brief study of basic metabolic pathways and formation of different secondary metabolites through these pathways- Shikimic acid pathway, Acetate pathways and Amino acid pathway.
b) Study of utilization of radioactive isotopes in the investigation of Biogenetic studies.
The document summarizes a study that assessed different aeration to non-aeration time ratios in reactors treating wastewater with high ammonia levels. Nitrifying bacteria populations were monitored using molecular methods like FISH and slot-blot hybridization. Results showed that longer non-aerated periods selected for Nitrospira over Nitrobacter. Measured nitrifier levels were sometimes lower than predicted by modeling, possibly due to undetected novel ammonia-oxidizing bacteria.
Effect of mixed species biofilm on corrosion of cast ironferibman
This document outlines an experimental study on the effect of mixed bacterial biofilms on corrosion of cast iron. The study aims to evaluate corrosion under mixed biofilms of denitrifying Pseudomonas aeruginosa and sulfate-reducing bacteria compared to individual biofilms. Materials and methods describe culturing the bacteria, preparing cast iron coupons, and electrochemical techniques like EIS and potentiodynamic polarization to characterize corrosion both with biofilms intact and after their removal. Preliminary results show the mixed biofilm reduces corrosion compared to sulfate-reducing bacteria alone, possibly due to interactions between denitrification and sulfate reduction pathways. Further experiments are planned to quantify biofilms and observe morphologies.
The document outlines an iGEM project that aims to detect and remove cadmium from the environment by engineering Bacillus subtilis to sense and sequester cadmium. It describes several subprojects involving building genetic circuits for cadmium sensing, stochastic switching of gene expression, cadmium sequestration using metallothioneins, and modeling the population-level effects of modifying the bacteria's life cycle. The overall goal is to render cadmium bio-unavailable by packaging it inside resilient bacterial spores.
Optimization of experimental protocols for cellular lysisExpedeon
In this project, we have compared existing sample preparation methods for proteomics studies against newly developed FASP method and our in-house developed SDS-TCA protocol. For our
preliminary studies, we have chosen a very well characterized soil microbe Pseudomonas putida.
Accessing genetically tagged heterocycle libraries via a chemoresistant DNA s...Laura Berry
Presented at the Global Medicinal Chemistry and GPCR Summit. To find out more, visit:
www.global-engage.com
Andreas Brunschweiger, an Independent Group Leader at TU Dortmund, discusses the limitations of DNA-encoded compound libraries (DELs) and getting around these.
GRAS proteins expression and purification Mesele Tilahun
The document summarizes the production and analysis of the GRAS protein Os-SCL7. It describes how the gene encoding the GRAS domain of Os-SCL7 was cloned and expressed in E. coli. The protein was then purified using nickel affinity chromatography and size exclusion chromatography. Sequence analysis revealed the protein is 378 amino acids with a predicted molecular weight of 41.5 kDa. Potential cleavage sites for specific proteases were also identified from the amino acid sequence.
Synthesis, spectroscopic, magnetic properties and superoxide dismutase (SOD) ...IOSR Journals
Three new ternary copper(II) complexes formulated as [Cu(HIda)(bipy)] 1; [Cu(HIda)(phen)] 2; [Cu(HIda)(dmp)] 3; where HIda =N-(2-hydroxyethyl)-2- iminodiacetic acid ; bipy = 2, 2’- bipyridine; phen = 1,10- phenanthroline; dmp = 2,9-dimethyl 1,10-phenanthroline, have been synthesized and characterized by partial elemental analysis, FAB-mass (m/z), EPR, UV-visible and CV measurements. The magnetic and spectroscopic data of all these complexes 1-3 indicate distorted octahedral geometry. The EPR spectra of these complexes in frozen DMSO solutions showed a single at g ca. 2. The trend in g-value (g||>g>2.0023) suggests that the unpaired electron on copper (II) has dx2–y2 character. The SOD activities of the complexes have been investigated. Antibacterial and antifungal activity of these complexes were also measured and discussed.
Similar to Microbial biosensors structure and mechanism (20)
How to Setup Warehouse & Location in Odoo 17 InventoryCeline George
In this slide, we'll explore how to set up warehouses and locations in Odoo 17 Inventory. This will help us manage our stock effectively, track inventory levels, and streamline warehouse operations.
ISO/IEC 27001, ISO/IEC 42001, and GDPR: Best Practices for Implementation and...PECB
Denis is a dynamic and results-driven Chief Information Officer (CIO) with a distinguished career spanning information systems analysis and technical project management. With a proven track record of spearheading the design and delivery of cutting-edge Information Management solutions, he has consistently elevated business operations, streamlined reporting functions, and maximized process efficiency.
Certified as an ISO/IEC 27001: Information Security Management Systems (ISMS) Lead Implementer, Data Protection Officer, and Cyber Risks Analyst, Denis brings a heightened focus on data security, privacy, and cyber resilience to every endeavor.
His expertise extends across a diverse spectrum of reporting, database, and web development applications, underpinned by an exceptional grasp of data storage and virtualization technologies. His proficiency in application testing, database administration, and data cleansing ensures seamless execution of complex projects.
What sets Denis apart is his comprehensive understanding of Business and Systems Analysis technologies, honed through involvement in all phases of the Software Development Lifecycle (SDLC). From meticulous requirements gathering to precise analysis, innovative design, rigorous development, thorough testing, and successful implementation, he has consistently delivered exceptional results.
Throughout his career, he has taken on multifaceted roles, from leading technical project management teams to owning solutions that drive operational excellence. His conscientious and proactive approach is unwavering, whether he is working independently or collaboratively within a team. His ability to connect with colleagues on a personal level underscores his commitment to fostering a harmonious and productive workplace environment.
Date: May 29, 2024
Tags: Information Security, ISO/IEC 27001, ISO/IEC 42001, Artificial Intelligence, GDPR
-------------------------------------------------------------------------------
Find out more about ISO training and certification services
Training: ISO/IEC 27001 Information Security Management System - EN | PECB
ISO/IEC 42001 Artificial Intelligence Management System - EN | PECB
General Data Protection Regulation (GDPR) - Training Courses - EN | PECB
Webinars: https://pecb.com/webinars
Article: https://pecb.com/article
-------------------------------------------------------------------------------
For more information about PECB:
Website: https://pecb.com/
LinkedIn: https://www.linkedin.com/company/pecb/
Facebook: https://www.facebook.com/PECBInternational/
Slideshare: http://www.slideshare.net/PECBCERTIFICATION
A workshop hosted by the South African Journal of Science aimed at postgraduate students and early career researchers with little or no experience in writing and publishing journal articles.
it describes the bony anatomy including the femoral head , acetabulum, labrum . also discusses the capsule , ligaments . muscle that act on the hip joint and the range of motion are outlined. factors affecting hip joint stability and weight transmission through the joint are summarized.
How to Add Chatter in the odoo 17 ERP ModuleCeline George
In Odoo, the chatter is like a chat tool that helps you work together on records. You can leave notes and track things, making it easier to talk with your team and partners. Inside chatter, all communication history, activity, and changes will be displayed.
Executive Directors Chat Leveraging AI for Diversity, Equity, and InclusionTechSoup
Let’s explore the intersection of technology and equity in the final session of our DEI series. Discover how AI tools, like ChatGPT, can be used to support and enhance your nonprofit's DEI initiatives. Participants will gain insights into practical AI applications and get tips for leveraging technology to advance their DEI goals.
Executive Directors Chat Leveraging AI for Diversity, Equity, and Inclusion
Microbial biosensors structure and mechanism
1. IN THE NAME OF GOD
Subject:
structure and mechanism of biosensor performance in toxic metal detection
Seyed Reza khedmati
P.H.D student of Microbiology
2. WAT IS BIOSENSOR?
Self-contained integrated device that capable of providing specific
qualitative or semi-qualitative analytical information using a biological
recognition element which is in direct-spatial contact with a transduction
element.(IUPAC,1998)
Biosensor: bio analytical System
7. WHAT KIND OF MOLECULES CAN BE DETECT?
• Protein
• Toxin
• Peptide
• Vitamin
• Sugar
• Metal ion
Cholera toxin Glucose Heavy metals
8. HEAVY METAL DETECTION
Heavy
metal
Detectio
n
by
Microorganism
s
Prokaryote
s
Eukaryotes
In Water
(Articles)
In Soil
(Articles)
A novel approach for rapidly and cost-effectively assessing
toxicity of toxic metals in acidic water using an acidophilic iron-
oxidizing biosensor
Love-wave bacteria-based sensor for the detection of heavy metal
toxicity in liquid medium
DETECTING BIOAVAILABLE TOXIC METALS AND METALLOIDS FROM NATURAL
WATER SAMPLES USING LUMINESCENT SENSOR BACTERIA
Immobilized bacterial biosensor for rapid and effective
monitoring of acute toxicity in water
Bioluminescent Bacterial Biosensors for the Assessment of Metal
Toxicity and Bioavailability in Soils
Assessment of heavy metal bioavailability in contaminated
sediments and soils using green fluorescent protein-based
bacterial biosensors
General
Articles
Heavy metal whole-cell biosensors using eukaryotic
microorganisms: an updated critical review
Whole-Cell Bacterial Biosensors and the Detection of
Bioavailable Arsenic
9. GENERAL DETECTION OF HEAVY METALS
Saccharomyces cerevisiae model among yeasts
Tetrahymena thermophila model for ciliates
Chlamydomonas model for microalgae
By Selecting The specific Reporter Gene
Heavy metal whole-cell biosensors using eukaryotic microorganisms: an updated critical
review
Reporter gene Advantages Disadvantages
β-Galactosidase(lacZ) Good stability Substrate dependent
Sensitivity depending on
substrate.
Low permeability
No ATP requirement Cellular lysis requirement
Eukaryotic luciferase
(luc)
Rapid response Substrate dependent
Very high sensitivity O2 and ATP
requirement
Low permeability and
stability Cellular lysis
requirement1
Green Fluorescent
Protein (gfp)
Good stability Moderate sensitivity
Substrate independent Lag-time for stable
fluorescence.
No ATP requirement Fluorescence after cell
death
No cellular lysis Autofluorescence
Comparison of the advantages and disadvantages of different reporter genes used in WCBs.
Part
1
10. Three main chelating molecules are involved in the
cellular
response to metals: glutathione (a tripeptide) , phytochelatins
(oligopeptides), and metallothioneins (proteins;
The two main reporter systems that are the best options for eukaryotic WCBs(whole-cell
biosensor) are:
the eukaryotic luciferase - the GFP(green fluorescent protein )
• CONCLUDING
• (i) eukaryotic microorganisms used as WCBs have certain advantages over prokaryotic cells. Among them, extrapolation of
the results to higher eukaryotic organisms is more reliable than using bacteria;
• (ii) inducible systems are more appropriate for designing heavy metal WCBs; (iii) the decision as to whether to use substrate-
dependent or independent reporters will be determined by the greater or lesser capacity for permeability of the substrate
through the wall or membrane of the cellular system used as WCB;
• (iv) in general, few WCBs are validated using bioassays with real environmental samples;
• (v) the biotechnology for using microalgae as WCBs is still underdeveloped, although these photosynthetic microorganisms
have a great potential as biosensors based on genetic constructs involving photosynthesis genes;
• (vi) ciliates are eukaryotic microorganisms that have a series of advantages over yeasts or microalgae for designing heavy
metal WCBs; and (vii) with regard to heavy metal WCBs for use in real environmental polluted samples, the capacity for
sensitivity of the biosensor is more important than its level of specificity to a metal.
11. WHOLE-CELL BACTERIAL BIOSENSORS AND THE
DETECTION OF BIOAVAILABLE ARSENIC
• Bioavailable Arsenic is able to penetrate the membrane of the bacterial biosensor
and trigger the detectable response, luminescence, which can be measured.
bioavailable arsenic can be estimated by this.
• Compounds found are arsenic trioxide, sodium arsenate, and arsenic trichloride.
• In gram-negative bacteria, the arsenic resistance gene remains inactive with the
absence of As(III) in the cell due to the binding of the ars operon repressor protein to
the promoter region of the gene. As(III) activates the system by binding to the
repressor protein and freeing the promoter region for transcription .The freed
promoter region is transcribed to produce various components of the mechanism such
as arsB, an arsenite-translocating protein that serves as a transmembrane efflux
channel. This protein functions either chemiosmotically, without an energy source, or
by ATP hydrolysis when coupled with arsA, an arsenite-specific ATPase. ArsC, the
enzyme arsenate reductase, is also transcribed to reduce As(V) to As(III), since As(V)
cannot pass through the arsB/arsA pump. ArsD is a regulatory protein for additional
control over the expression of the system and arsR is a transcriptional repressor (Figure
1) The mechanism varies slightly in gram-positive bacteria, which lack arsA and arsD.
12. Oraganisms by luxAB in plasmid
p1258:
Staphylococcus aureus
Escherichia coli
Attachment of As ion: Activation of
GFP & luminescence
Detect by :Fluorimeter & Luminometer
13.
14. BIOLUMINESCENT BACTERIAL BIOSENSORS FOR THE ASSESSMENT OF
METAL TOXICITY AND BIOAVAILABILITY IN SOILS
• Important factors
Bioavailability of heavy metals
PH of soil
organic matter and the clay content of soil
nature of microorganisms
texture and iron oxide concentration & binding of metals by cell walls or by
proteins and extracellular polymers, • formation of insoluble metal sulfides, •
volatilization, and • enhancement of export from cells
attachment factors
either adsorption to the polysaccharide coating or adsorption to binding sites such as
carboxyl, phosphate, sulfhydryl, or hydroxyl groups on the cell surface.
Other factors:
releasing specific compounds that form complexes with metals and by modifying soil pH
Gene resistance on plasmids and transposons
DETECTION OF HEAVY METALS IN SOIL
Part
2
15. • lux genes from Vibrio fischeri
• plasmid pI258 (cad A & cad C genes) from Staphylococcus aureus
• genes, zntA and zntR from Escherichia coli
• Genes of Arsenic ion’s resistance in Gram-negative bacteria
• chrA and chrB genes from Ralstonia metalliduran
In orther to using measurable signal :
inaZ coding for ice nucleation protein, lacZ coding for ß-galactosidase, gfp coding for
for the green fluorescent protein, lux genes coding for the bacterial luciferase system,
lue genes coding for firefly luciferase, and genes encoding for the enzyme catechol 2,3-
2,3-dioxygenase
16. • A green fluorescent protein (GFP)-based bacterial biosensor Escherichia coli DH5a
(pVLCD1) was developed based on the expression of gfp under the control of the cad
promoter and the cadC gene of Staphylococcus aureus plasmid pI258
• This mechanism increase detection of heavy metals in soil
To detect the bioavailable heavy metals in soil:
one approach is based on the use of bacteria that are genetically engineered so that a
measurable signal is produced when the bacteria are in contact with bioavailable metal ions:
reporter genes such as lacZ, lux, and luc to detect Cd(II)/Pb(II)
colorimetric enzyme assay and bioluminescence have been very successful as a reporter for
Cd(II)/Pb(II) detection
The gene for green fluorescent protein (GFP) from the jellyfish Aequoria Victoria for detection
Assessment of heavy metal bioavailability in contaminated sediments and soils
using green fluorescent protein-based bacterial biosensors
17. • In this research , researcher describe the construction of a
nonpathogenic Escherichia coli whole-cell biosensor for the detection of Cd(II), Pb(II),
and Sb(III) by employing red-shifted GFP (rs-GFP) as a reporter protein
. The sensor plasmid is based on the expression of rs-GFP under the control of the cad
promoter and the cadC gene of the cadA resistance determinant of Staphylococcus aureus
plasmid pI258
To make new detector plasmid for heavy metal’s:
Colonization and extraction of Plasmid pI258 isolated from S. aureus
PCR primers designed with either EcoRI (forward primer) or BamHI (reverse primer)
recognition sequence extensions and addeded
The resulting recombinant plasmid, pVLCD1 ,was transformed into E. coli DH5a by the
CaCl2 competent cell method
Schematic organization of the biosensor plasmid pVLCD1.
18. • The induction of the sensing system by a variety of metal
ions,
including As(III), Co(II), Cu(II), Fe(II), Hg(II), Mn(II), Ni(II), Sn(II), Cd(II),
Pb(II), Sb(III), and Zn(II) was studied by measuring the green fluorescence produced
For chemical analysis, concentrations of Cd(II), Pb(II) in water extracts of soil samples
were determined with inductively coupled plasma atomic emission spectroscopy analyzer
(PerkinElmer 3000SC, Norwalk, CT, USA).
Result:
Selectivity of the bacterial biosensor to metal ions. DH5a (pVLCD1) was treated with 1 mmol L1 of various individual metal ions or
mixtures of metal ions for 2 h. Induction intensity (in %) is defined as value of culture specific fluorescence (in SFI) with metal
treatment minus culture specific fluorescence (in SFI) of control then divided by culture specific fluorescence (in SFI) of control.
. Fluorescence of biosensor strain carrying pVLCD1
exposed to Sb(III). DH5a (pVLCD1) was treated with 1
mmol L1 of Sb(III) for 2 h at 37 C in LuriaeBertani (LB)
medium
19. Time-dependent induction of green
fluorescence with effectors. The DH5a cells
harboring the pVLCD1 plasmid were exposed
to 4 mmol L1
Cd(II), Pb(II), or Sb(III), and the specific
fluorescence intensity (in SFI) was determined
after different exposure periods
20. • Detection technique using a change of color, colorimetric, is widely used in the detection of
biomolecules, metal ions, and the presence of other compounds because the response can be
seen directly with the naked eye without the need for specialized instruments
• Immobilization is a technique that allows the bacteria can be used for long-term without
reducing its ability as bioreceptor
• Among the immobilization techniques, trapping method is one method which can be used for
cell immobilization.
• . In this method, the bacteria will be in a trap in the form of a matrix polymer
• example of matrix polymers: agarose, acrylamide, chitosan and alginate
Part
3
DETECTION OF HEAVY METALS IN WATER
Immobilized bacterial biosensor for rapid and effective monitoring of acute toxicity in water
21. IN THIS STUDY:
• E. coli was entrapped in the form of beads using calcium alginate
• Other materials as a reagent for sensing toxicant : Sodium alginate - calcium chloride -
Potassium ferricyanide -sodium chloride- ferric chloride - sulfuric acid, hydrochloric acid,
disodium hydrogen phosphate, sodium dihydrogen phosphate
Using :E. coli ATCC 25922
Culture & incubation & centrifuge & mix by calcium alginate
Absorbance value measured using spectrophotometer ( 600 nm)
Prussian blue was a colorimetric indicator of the process of
reduction of ferricyanide to ferrocyanide during bacterial
respiration with the addition of FeCl3 and can be measured
using spectrophotometer
22. • The relative activity of bacterial beads for monitoring
various
toxicants with different concentration levels (below Fig ) proved that
the higher the toxicant concentration, the smaller the response value. The higher the
toxicant concentration, the less the amount of K3[Fe(CN)6] converted to K4[Fe(CN)6]
during bacterial respiration process resulted in the amount of Prussian blue produced
was also decreased because toxicant inhibits bacterial respiration
23. Profile of reuse cycles of bacterial
biosensor beads for monitoring of
The stability of bacterial biosensor beads after
different storage times at 4 for monitoring toxicants
in water
24. • The ®re¯y (Photinus pyralis) luciferase (Luc) gene was used as a reporter in sensor strains
constructed.
• Luciferase catalyzes the oxidation of the heterocyclic substrate D-luciferin in the presence of
ATP producing visible light.
• The quanti cation of light emission . bioluminescence is one of the most sensitive means of
detection and it can be measured from living cells with a liquid scintillation counter, a
luminometer or even with X-ray ®lm
DETECTING BIOAVAILABLE TOXIC METALS AND METALLOIDS FROM
NATURAL WATER SAMPLES USING LUMINESCENT SENSOR BACTERIA
It presents schematically how the sensor bacteria work. In the absence of the inducing metal the expression of the
reporter gene is repressed, once the metal is added the expression of the reporter gene is induced and after substrate
addition (D-luciferin) the cells become luminescent, which can easily be measured
25. • Freeze-dried cells were reconstituted by two diffrent methods. Bacillus subtilis strains,
BR151(pTOO21) and BR151(pTOO24), and toxicity measurement strains Staphylococcus
aureus RN4220(pCSS810) and Escherichia coli MC1061(pCSS810) were reconstituted
• containing 0.5% casein hydrolysate for E. coli and S. aureus strains detecting mercury and
arsenite,
• containing 0.1% casein hydrolysate for the arsenite detecting B. subtilis strain
After addeding heavy metals:
The luminescence was measured with a microtiter plate reading luminometer, Luminoskan
(Labsystems, Helsinki, Finland)
26. RESULT
Spiked water samples. Metal sensing ability presented as induction coefficients of natural
water samples spiked with arsenite (AsOÿ
2 ), cadmium (Cd2+), lead (Pb2+) and mercury
(Hg2+).