OPPURTUNITIES IN ENVIRONMENTAL BIOTECHNOLOGY

MADE BY: MUKUL ATTRI SEC-S 9013 SUBMITTED TO: Ms. Sunishtha yadav

INDEX
HIGHLIGHTS OF CURRENT TRENDS IN ENVIRONMENTAL BIOTECHNOLOGY THE HOPE: A SUSTAINABLE FUTURE THE OPPURTUNITY: ENVIRONMENTAL BIOTECHNOLOGY RETRIEVING LOST ENERGY THE SCINCE BASE PROBING THE MICROBIAL COMMUNITIES ENVIRONMENTAL BIOTECHNOLOGY BENEFITS SOCIETY CENTER FOR ENVIRONMENTAL BIOTECHNOLOGY BIODESIGN INSTITUTE AT ARIZONA STATE UNIVERSITY WHAT MAKES A SUCCESSFUL ENVIRONMENTAL BIOLOGY A SUCCESS STORY:HYDROGEN BASED MEMBRANE FILM BIOREACTOR AN EMERGING REVOLUTION:MICROBIAL FUEL CELL BIOREMEDIATION OF RADIOACTIVE CONTAMINANTS BIOFILMS-THE GOOD AND BAD PHOTOBIOCATALYSIS MOLECULAR MICROBIAL ECOLOGY BIOHYDROGEN

HIGHLIGHTS OF CURRENT TRENDS IN ENVIRONMENTAL BIOTECHNOLOGY

Cleaning up polluted water Biodegrading toxins Protecting human health Generating power with bioelectricity Forging partnerships with industry Launching sustainable technologies
The Science Base Microbial Communities Ne

Research

THE HOPE: A SUSTAINABLE FUTURE

To safeguard the future for coming generations, we must conserve natural resources and rein in pollution. At the same time, we must create jobs to raise global living standards, including the living standards of those who currently suffer from lack of water, sanitation, education, and economic opportunity. How is it possible to conserve resources, reduce pollution, and promote dynamic economic growth? Sustainable development will allow us to meet our present needs without compromising the ability of future generations to meet their needs.

In Mexico, a child plays near a landfill, while his mother washes in a pool of rainwater.

As our population grows, our footprints must become smaller so that we do not leave a polluted world, stripped of resources for future generations. Using renewable resources, reducing pollution, and capturing energy resources from materials that we now throw away will transform society in ways that improve human life, now and in the future. The most precious resource in the 21st century will be water. Experts on the Middle East already acknowledge that water, not oil, will create conflict in that war-torn region. Like the Middle East, Arizona, too, is arid, and the states future prosperity depends on achieving water sustainability. Arizona is at the vanguard of a waterlimited era, but any region with a dense population, modern agriculture, or industrial activity is at risk. This is because fresh water is essential to our economic and physical survival, and, often, that water is in short supply. Increasing demand for fresh water forces society to use lowquality sources: i.e., wastewater, salty water, and polluted groundwater and river water. Low-quality sources must be treated to remove contaminants that affect public health, make water unsuitable for manufacturing, or give it a bad taste, odor, or color.

A renewable fuel source, bioelectric power, could fuel factories and reduce air pollution from combustion. In arid regions, water, not oil, will be the most precious resource. Above is the Egyptian desert and below an irrigated, Egyptian date palm plantation.

Highlights AS

Standing alongside water as a precious resource is energy. To provide a sustainable supply of energy in the future, we cannot rely on the primary sources of energy we use today. Now, we are extracting natural resources oil, coal, and natural gas that were accumulated over many millions of years. The earth cannot renew them as fast as we are extracting them. To be sustainable, our societys energy sources must come from renewable sources, instead of from fossil fuels. Oil priced greater than $50 per barrel is a harbinger of skyrocketing fossil-fuel prices in the future. The growing demands from China and India will accelerate the supply-demand tug-of-war. But, even at todays level of demand, pollution from combustion creates environmental problems that threaten sustainability. Energy generation from combustion is inherently inefficient. If we could run our cars and factories with no combustion, we could take a giant step toward sustainability. Renewable energy sources that do not require combustion are essential if we want to keep driving cars and running the turbines in our factories.

The Opportunity: Environmental Biotechnology

Advances in molecular biology, materials, and computing have opened up a new discipline, one that cuts across all of these specialties and that can help make sustainability a realistic goal. The name of this field is environmental biotechnology. It uses communities of naturally occurring microorganisms to improve the quality of water, air, and soil. Environmental biotechnology also captures pollutants and transforms them into energy. The ultimate strength of environmental biotechnology is that it eliminates the waste, potentially turning it into a resource. Microorganisms can remove every significant chemical contaminant, except for salt, from low-quality water. This is possible because naturally occurring microorganisms gain energy by oxidizing (removing electrons) contaminants while others gain energy by reducing (adding electrons) contaminants. These oxidation-reduction reactions turn toxic contaminants into harmless compounds, such as table salt. When microorganisms remove contaminants to benefit human society, they gain energy to grow and sustain themselves. Thus, environmental biotechnology creates a winwin scenario in which the microorganisms provide society a renewable water resource, while they gain what they need to live. Low-quality water may also contain resources such as energy, nutrients, and valuable metals. When captured and reused, these resources reduce societys demand for extracting them from the environment. However, if these substances are discharged into the environment, they cause environmental assaults, including oxygen depletion, eutrophication, and toxicity to humans and ecosystems. We can use environmental biotechnology to capture these resources, instead of having them pollute the environment.

Polluted water contains valuable resources that can be extracted to serve useful purposes. Dr. Bruce E. Rittmann, Director of the Center for Environmental Biotechnology at Arizona State University, stands in front of the CEB laboratory.

Environmental biotechnology can accomplish many goals that lead to a sustainable future. Environmental biotechnology has such promise because of the almost infinite versatility of microorganisms to find ways

to live; their ability to thrive in unusual conditions serves society. Where do we find these microorganisms? We find them living naturally in microbial communities. These busy little villages of microbes can only be seen with a microscope, but they are everywhere. Microbial communities are ecosystems that organize themselves when conditions are right. The environmental biotechnologist first understands the conditions that select for the right microorganisms and then creates a technology that sustains those conditions at a cost that society is able to pay. It is a perfect marriage of scientific research and engineering know-how.
Highlights A Sustainable Future Fresh Water & Energ

Retrieving Lost Energy

The greatest impact of environmental biotechnology may come from capturing the energy value of organic compounds in wastewaters, sludges, and agricultural products. Certain microorganisms naturally create valuable energy outputs as they biodegrade organic material. Some produce methane, or natural gas; others produce hydrogen gas, which can be used in a conventional fuel cell; and the newest discovery is that microorganisms generate electricity which can be used to power fuel cells. Were developing this discovery into a microbial fuel cell (MFC), which we think promises a clean, pollution-free way to power automobiles and run factories. Environmental biotechnology finds a way to harness these microorganisms to capture the energy outputs reliably and at a reasonable cost. In this way, environmental biotechnology yields a big payout to society. For example, converting the energy value in domestic sewage to electricity could save 2 million barrels of oil each year in the United States. Capturing the energy value from feedlot manure could be worth 6 million barrels per year.

A Sustainable FutureCapturing energy from feedlots could save millions of gallons of oil a year. Biohydrogen

The Science Base

Environmental biotechnology is truly a knowledge-driven technology. Once the right conditions are in place, the microbial community does the heavy lifting, accomplishing societys goals economically, simply, and safely. The first step toward success with an environmental biotechnology is being able to think like the microorganisms, which means performing scientific research to understand what the microorganisms do and what constitutes a good environment in which they can do it. The science core of environmental biotechnology is applying information from the science of microbial ecology. More specifically, the environmental biotechnologist needs answers to these fundamental questions of microbial ecology: What microorganisms are present? What can its members do? For example, can they destroy toxic chemicals or transform organic waste into a valuable energy source? What are its members actually doing? For example, is detoxification actually occurring? What are the interactions among the communitys members and with their environment? For example, do the members cooperate or compete? Do they require special spatial organization to function well? The environmental biotechnologist provides conditions so that the right microorganisms the ones that perform the right services thrive and stay within the community. This is the natural, ecologically based recipe for success with environmental biotechnologies.
A Sustainable Future Fresh Water & Energy The Opportunity Capturing Energy The Science Base Microbial Communities New Technology

Probing Microbial Communities

Thinking like the microorganisms demands a deep understanding of the microbial communities. Fortunately, we have powerful new tools to help us analyze these intriguing organisms. Among the tools we can use are molecular methods that probe the genetic information of the microorganisms in the community. By targeting different types of DNA or RNA, these tools can identify which microorganisms are present, what reactions they can perform, what reactions they are performing, and which physical and metabolic interactions are occurring. We know that microbial communities profoundly alter their environment by carrying out chemical reactions. Chemical tools measure the effects of microbial activity. Using these tools, we are able to understand how the microorganisms process chemicals to gain energy and grow (their biochemistry) and how those reactions alter the geochemical conditions of the environment in which the microbial community lives and works. Another set of tools includes mathematical models that systematically and quantitatively represent the community members, their reactions, and how they affect each other and their environment. Modeling uses computers, and it is a tool for integrating different types of knowledge. When we integrate information from all the tools, we gain a deep understanding of the biological and chemical activity in microbial communities.

In the image above, phase contrast (left) and fluorescent in situ hybridization (right) microscope images show the dense clusters of ammoniaoxidizing bacteria (red) and nitrite-oxidizing bacteria (green) within a larger aggregate. These bacteria came from a biological wastewater treatment plant. Model simulations, like the one below, are among the tools used to study microbial communities. This model simulates a biofilm structure with dense clusters of microorganisms. Such complex forms occur when the supply of food to support biofilm growth is restricted.

Hiring Energy

Environmental Biotechnology Benefits Society

Molecular microbial ecology and computer modeling are examples of the research bricks that go into creating a successful environmental biotechnology. However, to make advances in wateror energy-sustainability, these research bricks must be assembled into a building. The engineer designing this building is an environmental biotechnologist. Able to think like the microorganisms, the environmental biotechnologist creates systems that work for the microorganisms so that they can work for us. On page 12, the box entitled What Makes A Successful Biotechnology? shows what it takes to make microorganisms work for us. Any successful environmental biotechnology must work continuously in practical, scalable environments not just the lab, but in the real world. It must be reliable and cost- being translated into practical technologies. Both rely on modern materials and sound engineering. Two New Technologies The Membrane Biofilm Reactor (MBfR) has moved from the lab bench, to pilot projects, to field sites. Through partnerships with industry, this technology is being put it to work to clean up contaminated water. The MBfR reclaims low-quality water polluted with one or more oxidized contaminants, which cause serious health problems and are otherwise very difficult to remove . A second example is the Microbial Fuel Cell (MFC), which directly converts organic matter in wastes or renewable energy crops to electricity. It is still in the research phase, but moving forward rapidly. In both cases, the research needed to develop new biotechnologies, like the MBfR and MFC, must be deep, broad, and directed toward achieving success. The research integrates molecular microbial ecology, material science and engineering, quantitative modeling, and engineering design and optimization. Success comes about when an exciting scientific breakthrough, new materials, and good engineering are connected throughout research and development. Future
Fresh Water & Energy The Opportunity Capturing Energy The Science Base Microbial Co

The Center for Environmental Biotechnology

Integrating science-driven understanding with engineeringbased implementation is the greatest challenge in environmental biotechnology today. It is the challenge that inspires the new Center for Environmental Biotechnology in the Biodesign Institute at Arizona State University.

Use-Inspired Research for a Better Future Research at the Center for Environmental Biotechnology is use-inspired, which means that it is directed towards providing an essential service to make society more sustainable. All research in the Center is guided by these three principles: We employ leading-edge research tools including molecular microbial ecology and modeling so that we can develop a fundamental understanding of microbial communities and think like the microorganisms. We apply modern materials including membranes and nano-materials and good engineering strategies to create systems that work for the microorganisms so that they work for us. We partner with private and public companies, individual investors, and the public sector to commercialize our technologies and get them into the marketplace.

This breadth of activities at the Center of Environmental Biotechnology and its use-inspired focus are what make it unique as a research center. Researchers in the Center constantly integrate tools from many disciplines and are stimulated by real-world goals. The Center for Environmental Biotechnology is uniquely able to create new knowledge about microbial communities and apply that knowledge.
Highlights A Sustainable Future Fresh Water & Energy The OpportunitUse

The Biodesign Institute at Arizona State University

Dr. Michael Crow, President of Arizona State University, has launched an initiative to create the New American University, a university that advances the well being of all the citizens of Arizona, as well as the nation and the world. Reflecting that vision, ASU has made sustainability and the use of biology in the service of society its top priorities. The Biodesign Institute at ASU was created as a bold new approach to make those dreams a reality. The Biodesign Institute uses natures design as inspiration for highly inter-disciplinary discovery, development, and delivery of innovative solutions to societys urgent problems of health, security, environment, and sustainability. The Center for Environmental Biotechnology is located within this state-of-the-art facility.HighlightsA Sustainable Future The S-Curve of New Technology Any new technology takes time before it gains acceptance, and while this talk of microbes may seem far off, in fact, microbial activity has been taking care of our waste and helping clean up water for a long time. Below, you will find three examplesold and newof this technology.
Table 1-Technology That Relies on Microbial Activity

What Makes a Successful Environmental Biotechnology?

Success with an environmental biotechnology begins with managing the communitys structure so that its function does the job by providing a service. Although the structure-function relationship is the foundation for success, it is only the beginning. A truly successful environmental biotechnology also must meet these -world criteria: It must be practical at a large scale. The relevant units normally are huge: millions of gallons per day for flow tons/day for mass. It must operate reliably for continuous use. We cannot turn off the flow if something goes wrong. We cannot get a time out if the conditions change. It must be economical to build and operate. Environmental biotechnology provides essential services that cannot be priced outside societys reach.

A Success Story :The Hydrogen-Based Membrane Biofilm Reactor

One example of a new biotechnology used to treat low-quality water is the hydrogen-based membrane biofilm reactor (MBfR), an environmental biotechnology that Dr. Rittmann has taken from fundamental research through commercialization activity. In the MBfR, we deliver hydrogen gas (H2) to microorganisms by allowing it to diffuse through the walls of special membranes that H2 bubbles from forming. The microbial community grows naturally as a biofilm on the outer wall of the membranes, by removing electrons from H2 and transferring the electrons to one or more oxidized contaminants in the water.

The oxidized contaminants include many new water contaminants whose harmful effects have recently been discovered. These include perchlorate, chromate, and selenate. Reducing these contaminants renders them harmless or easily removed from the water by common watertreatment methods. The scientific breakthrough behind the MBfR is the discovery that bacteria that oxidize H2 are able to reduce almost any oxidized contaminant. This breakthrough is translated into an effective technology by having the biofilm grow on the outside of a special membrane that does not allow bubbling of the H2. The combination of a biofilm on a no-bubble membrane is the breakthrough that makes microbial use of H2 reliable, efficient, safe.
Highlights A Sustainable Future Fresh Water & Energy The Opportunity

Capturing Energy The Science Base

At right, a greatly magnified photo of a biofilm (red) shows the growth on the membrane wall.

, a schematic drawingshows how hydrogen diffuses through the wall. A biofilm grows naturally on the walls surface.

An Emerging Revolution:The Microbial Fuel Cell

A revolutionary new environmental biotechnology the Microbial Fuel Cell (MFC) turns the treatment of organic wastes into a source of electricity. Bacteria growing as a biofilm on an electrode in a fuel cell oxidize the organic pollutants and transfer the electrons to the electrode, into an electrical circuit, and eventually to oxygen at a second electrode. .

In the future, cars that now consume fossil fuel could be powered by microbial fuel cells. A new source of energy called bioelectric power is being developed, thanks to the researchers working on the microbial fuel cell.

HighlightsMicrobial Fuel Cell

The MFC is revolutionary for three reasons. First, it makes the treatment of organic pollutants a direct producer of electricity, not a consumer. Second, it expands fuel-cell technology to use renewable organic materials as a fuel; conventional fuel cells use hydrogen gas, which is today produced from fossil fuels. Furthermore, the MFC can use organic fuels that are wet, the usual form for wastes and fuel crops. Third, the MFC, by operating at ambient temperature, can double to triple the electricity-capture efficiency over combustion, while eliminating all the air pollution that comes from combustion. The scientific breakthrough leading to the MFC is the recent that some bacteria can transfer electrons into an electrode and create electricity. This breakthrough is translated into a technology by using modern membrane and electrode materials that are compatible with biofilm growth and operation at ambient temperature.Sus

Like a conventional fuel cell, the microbial fuel cell has an anode and cathode, but the fuel source can be renewable organic material. Colonies of microbes called biofilms live on the anode and catalyse free electrons from the organic fuel.

Rad Highlights A Sustainable Future Fresh Water & Energy The Opportunity Capturing Energy The Science Base Microbial Communities New Technology Use-Inspired Research The Biodesign Institut

BIOREMEDIATION OF RADIOACTIVE CONTAMINANTS

Molecular Microbial Ecology

Biohydrogen

Among the most toxic chemicals of global concern are the radionuclides found in contaminated waste streams and groundwaters. For example, former nuclear-weapons facilities are severely contaminated with plutonium, uranium, and neptunium because of past practices that released mixtures of the radionuclides and other organic and inorganic into adjacent soil and

groundwater. The greatest potential for cleanup of such sites is in situ bioremediation, which exploits the reactions of microorganisms to directly or indirectly alter the chemical form of the radionuclides, rendering them immobile and less toxic. The microorganisms interact directly with the radionuclides by catalyzing chemical redox transformations. The microorganisms act indirectly by producing acids, bases, and complexing ligands that react with the radionuclides.

Radionuclide bioremediation is a complicated situation, since the fate of radionuclide depends on so many different reactions, which proceed through multiple steps and at vastly different rates. Such a complex scenario can be understood and controlled only by using mathematical modeling. Such a unique biogeochemical mathematical model, CCBATCH, was developed by

Dr. Rittmann and colleagues to connect all the different types of reactions that control the fate of radionuclides and a large range of metals. Current research is focused on applying CCBATCH towards bioremediation of plutonium, one of the most toxic materials. To create a clean environment for all the people around the world, it is necessary to mitigate waste problems left behind as legacies of dangerous past activities, such as making nuclear weapons. When understood well, such as with CCBATCH, in situ bioremediation has great promise to deal with one of the most difficult challenges -- groundwater and soil contamination by radionuclides, such as plutonium.
Highlights

A Sustainable Future Fresh Water & Energy The Opportunity

Capt

Biofilms The Good and the Bad

Biofilms refer to the assemblages of microorganisms and biologically produced macromolecules attached to a surface. Biofilms are everywhere; they are found on surfaces in contact with moisture, such as water lines and soft tissues. Biofilms can be categorized as good or bad. Good biofilms are responsible for nutrient cycling in nature and for biodegradation of pollutants in water-treatment technology. Engineers often wish to develop a dense and firmly anchored biofilm in treatment technologies. Bad biofilms, or biofouling, develops on surfaces in which microorganisms are not desired. Biofouling in a water line may interfere with the systems circulation or may harbor pathogens. Biofouling is a major source of infection in humans and plants, therefore we want to prevent or remove biofouling biofilms. During the formation of biofilms, microorganisms usually encase themselves in extracellular polymeric substances (EPS) or slime matrix, which improves microbial adhesion to the surface and to other cells. EPS in biofilms protect microbial cells from biocides and also increase tolerance to physical and chemical disturbances. EPS clearly plays a role in making the biofilm firmly attached or easily removed.

PHOTOBIOCATALYSIS
Biological processes are the overwhelming choice for wastewater treatment due to their ease of use, excellent performance, and efficiency. However, when challenged with toxic organic compounds, these processes can be ineffective and in some cases fail, resulting in discharge of harmful pollutants into the environment. Highlights Advanced oxidation processes (AOPs), such as photocatalysis, have wide-ranging applicability, are not susceptible to process upsets from toxic inputs, but generally are not cost-effective and not practical options for many real situations. By combining these two technologies, a process called photobiocatalysis, we take advantage of the benefits of each, while minimizing their drawbacks. Traditional work on coupled chemicalbiological treatment focused on sequentially coupled systems, or those that have the chemical and biological processes in separate stages, but these systems suffer from the indiscriminate nature of advanced oxidation, which results in a large range of products, including those that are too oxidized, toxic themselves, or unavailable for biodegradation. This situation could be improved by combining the two operations into a single-stage, called intimate coupling, whereby bacteria are in close proximity to advanced oxidation, and can therefore remove biodegradable products as they are formed, focusing chemical oxidant on the non-biodegradable fraction. We achieve intimate coupling by using a photocatalytic circulating-bed biofilm reactor, or PCBBR, shown here, which exploits biofilm carriers to hold and protect the bacteria from harmful advanced oxidation and toxic compounds, but places the bacteria as close as possible to the advanced oxidation so that the biodegradable products are removed as soon as they are produced, focusing the chemical oxidant on the nonbiodegradable fraction. The PCBBR technology offers the potential to efficiently and thoroughly treat many toxic wastewaters including those contaminated with halogenated aromatics, endocrine disrupting compounds, munitions, and a wide range of harmful industrial inputs including pharmaceutical wastes and textile dyes, resulting in a cleaner water being discharged to the environment, a healthier environment, and a healthier world for us to live in.

Molecular Microbial Ecology

The scientific core of environmental biotechnology is microbial ecology, a scientific discipline dedicated to understanding complex communities of microorganisms: what microorganisms are present, what is their metabolic potential, what part of the potential are they realizing, and how they interact with each other and their environment. Fundamental scientific research In microbial ecology provides us with a deep understanding of how the complex communities work. In short, we seek to be able to think like the microorganisms.

Fortunately, we have powerful new tools to help us analyze these intriguing organisms and their community organization. Among the tools we can use are molecular methods that probe the genetic information of the microorganisms in microbial communities. By targeting genomic DNA, we can identify which microorganisms are present and what reactions they can perform; through the use of RNA-based techniques we can identify the members in the community that are actively growing and what reactions they are performing. Using in-situ techniques, we can also investigate the metabolic interactions that take place among microorganisms in a mixed community. Applying these molecular tools to understand microbial communities is called

molecular microbial ecology, and it is a critical research strength of the Center for Environmental Biotechnology. In particular, the Center has exceptional capabilities to investigate changes in microbial structure through the use of molecular techniques that target the 16S rRNA gene and the 16S rRNA, such as denaturing gradient gel electrophoresis (DGGE), real-time PCR, and fluorescent in situ hybridization (FISH). DGGE is especially powerful for doing detective work to identify important, but uncharacterized strains. Real-time PCR is powerful for quantifying the different types of microorganisms. FISH allows us to do 3-dimensinal visualization of the communities and understand the interactions among different strains. The Center also targets specific metabolic genes and their expression to investigate the role of critical enzymes in detoxification and energyinvestigate the role of critical enzymes in detoxification and energygenerating reactions occurring in natural or engineered techniques including reverse transcriptase PCR, RNA or cDNA microarrays, and real-time PCR to quantify over- and underexpression of specific genes and for microarray validation. Identification of specific gene-targets will allow us to investigate critical factors affecting the performance of natural or engineered microbial systems. The expression of such critical genes can be used to monitor and evaluate the success of engineered processes. In summary, the Center for Environmental Biotechnology employs a range of tools from molecular microbial ecology so that it can understand complex microbial ecosystems, allowing us to think like the microorganisms.

Biohydrogen, a Renewable Source for Clean Energy

Hydrogen is the proposed fuel of the future. The reason for this hope for hydrogen is the development of efficient fuel cells that convert the energy in hydrogen to electricity with high efficiency and zero emissions of air pollutants. However, the future for hydrogen is clouded. Although hydrogen itself is very clean and attractive, almost all of the hydrogen produced at the present time comes from non-renewable fossil sources, such as natural gas and coal. This nonrenewable cloud over hydrogen can be lifted by biohydrogen. Biohydrogen refers to hydrogen produced by algae, bacteria, or biological components of these organisms. These organisms use renewable biomass or sunlight to produce hydrogen. As our society strives for renewable hydrogen, biohydrogen will be a main alternative. Furthermore, it integrates waste treatment with clean-energy production from renewable sources. For example, biohydrogen be used to produce electricity in a fuel cell, and it also is a special electron donor for bacteria in treatment processes used for reduced contaminants (such as the hydrogen-based membrane biofilm reactor). Although microorganisms can produce biohydrogen by various processes, fermentation is the simplest process and the one we are exploring. Fermentation is the essential

first step in any process to recovery energy from biomass. Biomass is made up of complex organic molecules. Fermentation generates a mixture of simpler molecules: organic acids, alcohols, and hydrogen. Thus, a great advantage of fermentation is fast degradation of solids and other complex organics found in wastes and agricultural products. On the other hand, fermentation today converts only about 15% of the energy to hydrogen. While fermentaion is fast, it is not yet efficient for capturing the energy value of biomass to hydrogen. EBs goal is to increase the biohydrogen yield to around 85%. To do this, we are investigating a multi-faceted research agenda that involves two complementary strategies. The first strategy involves controlling the microbial ecology in the fermentation process so that electrons and energy flow to hydrogen instead of to other end products. The second strategy involves directing the electrons and energy in the other fermentation end products.