2011 Science Report

2 Great Lakes Bioenergy Research Center

Our MissionOur mission is grand, but simply stated: to perform the basic research that generates technology to convert cellulosic biomass to ethanol and other advanced biofuels.

Our RoleIn order to focus the most advanced biotechnology-based resources on the biological challenges of biofuel production, the Department of Energy (DOE) established three Bioenergy Research Centers (BRCs) in September 2007. The Great Lakes Bioenergy Research Center (GLBRC) is led by the University of Wisconsin-Madison, with Michigan State University as the major partner. Additional scientific partners are DOE National Laboratories, other universities and a biotechnology company. GLBRC’s researcher expertise covers a wide array of disciplines, from microbiology to economics and engineering. Each Bioenergy Research Center is pursuing the basic research underlying a range of high-risk, high-return biological solutions for bioenergy applications. Advances resulting from the BRCs will provide the knowledge needed to develop new bio-based products, methods and tools for the emerging biofuels industry.

Our MembersPacific Northwest National Lab

University of New Hampshire

Lucigen Corporation

Illinois State University Oak Ridge National LabUniversity of Missouri–Columbia

Iowa State University

University of Toledo

Cornell University

University of Wisconsin–Madison

Michigan State University

University of British Columbia

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Our researchers are investigating a variety of bioenergy crops that could fuel our trips around town, across the country or around the globe. We are beginning efforts to make biofuels from both crop residues and dedicated energy crops to improve the energy efficiency, cost and sustainability of the entire process. With this big picture in mind, our scientists are exploring ways to provide fuels that are capable of burning in a variety of engines.

We can now deconstruct corn stalks into a sugar stream and feed that material to microbes and catalysts to make fuel. By understanding the conversion and fuel synthesis process at a molecular level — and seeing the parts work in real time — we can identify the elements that can be improved to increase the efficiency and sustainability of this conversion.

We also know that cellulosic biofuels will be more easily adopted by consumers if they connect as seamlessly as possible with the large and complicated infrastructure that connects pipelines, tankers, trucks and even jets.

Consequently, research within our Center considers all factors that go into developing cellulosic biofuels. Our strategy, call it comprehensive, holistic or “field to pump,” integrates numerous approaches and disciplines to improve cellulosic biofuels production economically, environmentally and energetically.

By harnessing the expertise of scientists and staff to solve Center-wide goals, we have evolved into a tightly knit research community. And our collective effort is paying dividends. We are generating new technologies (~40 patents in the works), publishing trans-disciplinary research (almost 300 papers) and training aspiring bioenergy scientists

(from undergraduates to the next generation of faculty members or industry leaders).

Given the path we are traveling, the expert team we have assembled, and the insight that our collaborative approach affords us, the Great Lakes Bioenergy Research Center is well positioned to generate basic science breakthroughs that will help make cellulosic biofuels a cost-effective, energy-efficient and sustainable substitute for fossil fuels.

4 Integrated Research Creates Broader Impact

5 Our Energy Opportunity6 Plants8 Deconstruction

10 From Field to Fuel

12 Conversion14 Sustainability16 Technology Transfer18 Education & Outreach

20 Contact Us

As you read our 2011 Science Report, you will get a glimpse of the high-quality research that is underway along every stop in our biofuels research pipeline—from growing plants to converting sugars to generating fuels.

Great Lakes Bioenergy is Gaining Momentum

Contents

Tim Donohue

Director, Great Lakes

Bioenergy Research Center

“By harnessing the expertise of scientists and staff to solve Center-wide goals, we have evolved into a tightly knit research community. And our collective effort is paying dividends.”

4 Great Lakes Bioenergy Research Center

Year

4Q

1-Q

3Ye

ar 3

Year

2Ye

ar 1

Journal Impact Factor19+

13-18

10-126-9

4-5

2-3

<2new journals, book chapters

Collaboration

2 Labs

3 Labs

4+ Labs

1 Lab

Journal Impact Factor is a measure of the average number of citations of papers published in a journal—which serves as a way to measure the influence of that journal within the scientific field.

Since GLBRC was founded in 2007, research output in peer-reviewed journals has grown at a

steady rate. In the figure below, the gradual shift from black to red shows that as the

publication rate has grown, more publications were authored by

researchers from multiple labs within the Center. This reflects the

trans-disciplinary collaboration that is a result of bringing

400 researchers and sta� together in a Center

model.

The past four years have been an exciting time for the researchers at Michigan State University participating in the Great Lakes Bioenergy Research Center. Our partnership in the GLBRC has provided opportunities to collaborate across campuses and across disciplines that are unprecedented. While research collaborations between scientists and engineers are no longer rare in many fields, a number of projects in the Center have jumped much larger disciplinary boundaries.

Within the GLBRC we have chemical engineers, economists, modelers and ecologists working together on a very challenging problem: how to deploy new bioenergy technologies in the rural environment. Sustainable deployment of new bioenergy technologies, ranging from agricultural practices to end use, has emerged as a

critical national issue. The GLBRC is uniquely positioned to lead the way to solutions. These

solutions will require the diverse insights from many different disciplines, as well

as innovative communication and educational outreach programs.

Innovation occurs most readily when different

fields overlap, bringing together new ideas

and technologies.

By using an integrated approach to biofuels research, GLBRC is leveraging this innovation to generate top-tier research publications, new biofuel technologies and strategic partnerships.

Collaborative approaches to science become contagious once investigators learn how advantageous they can be in problem solving. At MSU, we have leveraged advances from the GLBRC

to expand collaborations even further. GLBRC Researcher Bruce Dale and MBI won a $4.3 million DOE grant to build an engineering scale plant for the ammonia fiber expansion pretreatment known as AFEX.™ This collaboration will test design factors for commercial scale-up, as well as produce sufficient AFEX™ material to evaluate it as an animal feed in a full-scale trial with dairy cattle. Working with animal nutritionists and veterinarians, we have gone beyond our bioenergy origins to explore the possibilities for having both fuel and food. Collaborations lead to very interesting places!

Integrated Research Creates Broader Impact

Doug Gage

Director, MSU BioEconomy

Network, Office of the VP for

Research and Graduate Studies

“By using an integrated approach to biofuels research, GLBRC is leveraging this innovation to generate top-tier research publications, new biofuel technologies and strategic partnerships.”

1. AFEX™ is a trademark of MBI.

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1http://www.eia.gov/totalenergy/data/annual/showtext.cfm?t=ptb0201e2https://bioenergykdf.net/content/billiontonupdate3http://www.eia.gov/energy_in_brief/foreign_oil_dependence.cfm

4http://www.eia.gov/tools/faqs/faq.cfm?id=36&t=65http://www1.eere.energy.gov/biomass/ethanol_myths_facts.html6http://www.newyorktransportation.com/info/empirefact2.html

Our Energy OpportunityIn 2010, the transportation sector of the United States consumed 27,425 trillion BTUs of fuel, 95.996% of which were from fossil fuels.1 The biomass resources identified by Department of Energy’s Billion Ton Study (2011)2 could be used to replace approximately 30 percent of the nation’s current petroleum consumption with clean, renewable biofuels.

Transportation Fuel Consumption

In 2010, the United States imported approximately 3.445 billion barrels of oil from foreign countries,3,4 approximately 51% of U.S. consumption.

In comparison to gasoline, ethanol made from cellulose and produced with power generated from biomass byproducts can result in an 86 percent reduction in greenhouse gas emissions.5

Foreign Oil

Fuel Gasoline Corn-grainEthanol

19%Reduction

CellulosicEthanol

86%Reduction

Fossil Fuels BiofuelsToday

Fossil Fuels Biofuels

67.19%Tomorrow’s Opportunity

Greenhouse Gases

Transitioning to renewable biofuels

keeps energy spending closer to home.

In 2010, the United States imported 547,864,671,000 liters of oil.4

That’s enough to fill the Empire State Building6 523 times.

Other

Picture or 2-3 pictures

6 Great Lakes Bioenergy Research Center

Productivity

Durability

Convertibility

First Stop: PlantsAs they explore how to degrade plant

biomass into simpler sugars or coax crops into growing more biomass per

acre, researchers at the Great Lakes Bioenergy Research Center (GLBRC)

are identifying the characteristics essential for better biofuels feedstocks.

The Center’s approach for creating better plants for biofuels is varied, from plant breeding to mapping the genome of developing maize to a quantitative analysis in switchgrass genetics. The ability to peer into the plant’s code, through the use of cutting-edge genomic tools, is available to researchers through a partnership with the U.S. Department of Energy Joint Genome Institute.

“We’re using genomics to speed up the selection process in plant breeding,” says Michael Casler, GLBRC researcher and USDA Agriculture Research Services geneticist. “It takes a long, long time to produce a new variety. So, it’s beneficial for research to shorten that period of time.”

Switchgrass is often mentioned among biofuel feedstock contenders, and two types of switchgrass are native to the United States—upland and lowland varieties. Adapted to the frigid falls and winters of the northern United States, the upland plants mature earlier. But, the downside to flowering early is that this variety accumulates less biomass. The lowland plants, on the other hand, have adapted for longer summers by flowering substantially

later in the season. They are fully mature months after the upland plants, and this allows them to grow larger and produce more biomass that can subsequently be converted to fuels.

Casler’s challenge? Combining the best characteristics, cold tolerance and maximum biomass, into a new variety.

“Our data says that we can increase biomass yields by 30 percent, just by shifting to the lowland plant type,” Casler says. “But, you can’t just shift to using that plant everywhere. The late flowering plants don’t survive very well in the North.”

After planting thousands of individual plants, Casler spends a year watching, waiting and hoping that some plants will survive for the next round of cross breeding.

Monitoring each generation of a trait through traditional plant breeding takes a substantial amount of time, which is why researchers often use faster-growing, genetically similar plant surrogates to speed up the process. By watching the development of the trait and tracking the results in so-called model crops, the team can determine how to activate the trait in

Plants

Our researchers are identifying important genetic traits that could make bioenergy crops like switchgrass more productive, more durable in harsh climates and more easily digested for conversion to biofuels.

72011 Science Report

another species.Model crops allow researchers to

monitor traits and genetic populations on much shorter life cycles, typically four to eight weeks.

“We’ve been able to use a model crop system to not only get faster insights into what genes and traits might be important, but also what mechanisms might be important,” says Kathryn Richmond, GLBRC’s director of enabling technologies. “So, when we move back to switchgrass or another similar species, we get a jumpstart.”

“The Center uses Brachypodium as a model crop for switchgrass because it is a grass species and shares a tremendous amount of genetic similarity,” Casler says. “When we find a gene that causes a desirable effect in Brachypodium, there’s a high chance of finding a similar gene in switchgrass and using it to create the same effect.”

Genomics helps researchers find and mark these desirable traits so they can track their progress in future generations and determine if the effect is maintained.

“The benefit is that we are selecting based on the markers for several generations. We can do that in seedlings, and we can accomplish the whole process in one year,” Casler says. “Whereas, if you select for a field-based trait, that process can take up to five years.”

After desirable traits have been selected

and hybrid plants bred, researched and analyzed, field trials begin to ultimately determine commercial viability of the new variety.

“For some species it’s straight forward to go from lab to bench to greenhouse to field,” Richmond says. “For other species it’s a more onerous and cautious step, and more safeguards need to be involved. In most cases we are cutting new ground here. We want to make progress safely and maximize the benefit for everyone.”

Moving Biofuels Forward with Genomics

Genomics is the study of genomes. It’s the investigation of a global cellular system that produces RNA, controls

gene expression and regulatory events, and ultimately determines an organism’s developmental process.

In the last five years, genomics research and technology has advanced rapidly. Then, the most daunting part of the process was sequencing an organism. Now, through advanced sequencing techniques, data can be generated quickly but takes significantly longer to review.

“In a couple of weeks we can collect data that could take us anywhere from months to a year to fully analyze,” says Richmond. “For research purposes, too much information and data is a great problem to have. Now we (and the rest of the field) are turning our sights on how to best visualize and mine this abundance.”

The GLBRC collaborates with the Joint Genome Institute, who receives funding from the U.S. Department of Energy specifically to secure a percentage of its capacity for bioenergy research.

In genomics, the idea is to be constantly improving the organism you are working on and studying. For the GLBRC this includes increasing plant biomass yields or developing novel ways to degrade biomass efficiently.

“Every time you turn around there’s been a breakthrough or new advancement,” Richmond says. “The JGI has really done a phenomenal job keeping up with the times, and our entire Center has benefitted in some way from this partnership.”

-E.A.

At the Arlington Agricultural Research Station in Wisconsin, GLBRC researchers are conducting field trials to evaluate how crops like corn stover, switchgrass, Miscanthus, poplar or native prairie would stack up as potential bioenergy cropping systems.

As his GLBRC colleague Michael Casler focuses on how to make switchgrass more productive, UW-Madison Biochemistry Professor John Ralph (pictured) and his team are working to create Zip-Lignin,™ a new technology that could more easily break apart one of the toughest compounds within plant cell walls.

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8 Great Lakes Bioenergy Research Center

Deconstruction

Second Stop: ProcessingJust as people need to chew food to

better access and digest the nutrients inside, mechanical and chemical

pretreatment of plants disrupts the cell walls and allows access to the

sugars within. Using ammonia, heat and pressure, a pretreatment method known

as AFEX™ (ammonia fiber expansion) blasts open cell walls, allowing enzymes easier access to the sugar polymers that

make up plant cellulose. Enzymes then break polymers apart into simple sugars

for conversion to biofuel.

“We’ve come up with a less costly way of doing AFEX™ that we think is ready to commercialize,” says Bruce Dale, GLBRC deconstruction lead and MSU professor of chemical engineering and materials science. Improvements to the AFEX™ pretreatment process have also reduced the need for costly enzymes by a factor of three. And if the team can push the technology further, accomplishing another three- or four-fold reduction, enzyme cost would no longer be a limiting factor in biofuel production.

As Dale’s team tinkered with different approaches to implementing this technology, they found that a modified approach to AFEX™ actually changed cellulose into a slightly different form that is five times easier for enzymes to break apart.

“We can understand in a much deeper way now how the AFEX™ process works, how it operates to produce digestible biomass,” Dale says. “Because we know that, we can do a much more rational job of picking the enzyme cocktails.”

Third Stop: EnzymesIn GLBRC’s early days, UW-Madison

Bacteriology Professor Cameron Currie’s work with leaf-cutter ants shed light on how these remarkable insects actually grow food—in one of the world’s oldest instances of farming—by tending leaves that provide nutrients for a strain of fungi that is the ant’s dietary staple. Along the way, Currie discovered something else: the ant’s nests are home to a number of previously unknown microbes whose enzymes may help break down the leaves. Currie recognized this property as a potential asset in the attempt to break down cellulose for biofuel.

That research has given Currie insight into the way cellulose-degrading microbes like bacteria or fungi work. One thing he’s seen so far is that microbes rarely go it alone. “Microbes in nature do not occur in isolation,” says Currie. “They do not break down plant biomass in a pure culture. In many systems, like the ant system, you have increased success and ability to compete with other organisms through beneficial symbiotic associations.”

At GLBRC, engineers and enzymologists are collaborating to design new ways of deconstructing plant biomass into its component sugars. Alkaline pretreatment strategies like AFEX™ are successful at loosening the tightly-packed cell wall components to increase access for enzymes that further break down cellulose into simple sugars.

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He predicts that within many of these symbiotic systems, combinations of microbial organisms are each producing different enzymes, and that these enzymes each play a part in the efficient breakdown of plant material.

The leaves and stalks of potential bioenergy plants are comprised of large quantities of cellulose, the most abundant organic compound on the planet. Cellulose is a polysaccharide, a long chain of tightly linked sugar subunits that must be broken down into simple sugars before they can be processed into biofuel.

The ability of enzymes to grow on pure cellulose and break it down into simpler sugars is a very unique capability—one that helps Currie and other GLBRC researchers substantially narrow the field of research subjects.

“If taking apart cellulose was easy, there wouldn’t be any trees around,” says Brian Fox, a UW–Madison professor of biochemistry who co-leads GLBRC’s deconstruction research.

From the 3,000 contenders that Currie and his team have sitting in cold storage, only 50 pass the cellulose test.

After making the first cut, Fox wants to know more about how the remaining bugs will function, including how they hold up in hostile environments. Are they picky, or can they withstand a variety of conditions?

After scouring natural environments and narrowing down the potential players, GLBRC deconstruction researchers have focused their attention on a novel microbe called Streptomyces sp. ActE (ActE).

Isolated from the community of wood-boring wasps, ActE seems to have an evolutionary advantage that makes it great at breaking down cellulose.

“The wood wasp is an interesting thing for us because it’s very specific,” Fox says. “For ten million years, they have been attacking pine trees, offering the potential to developed a specialized approach to deconstructing woody materials.”

From the start, GLBRC researchers realized that this bug had potential. Performance measures have shown that ActE is just as capable at breaking down cellulose as T. reesei, a fungus that is used heavily by industry as a source of biofuels enzymes.

Digging into ActE’s unique characteristics has surfaced an interesting discovery: ActE uses oxygen to metabolize cellulose.

“This realization is a brand new area in the whole of biofuels research. It’s a new paradigm,” Fox says.

Now that Fox’s team is intently focused on ActE, they’re using genomic tools like sequencing, microarrays and proteomics to learn why this bug seems to do its job so well. Since ActE secretes its proteins, studying how individual enzymes function is relatively simple.

Once specific ActE enzymes are isolated, Fox can look at each one and measure its activity, stability and impact on the overall effectiveness of the enzyme cocktail. “If protein X is the weak link, that’s a target for engineering.”

Currie and Fox are working to deliver newly characterized enzymes to other GLBRC researchers who study chemical pretreatment methods, solve three-dimensional crystal structures to better understand how the enzymes work and use a robot to create enzyme cocktails that release sugars from plant biomass. This robotics platform, GENPLAT, runs through an impressive 96 tests at once, allowing Jonathan Walton, MSU plant biologist and GLBRC colleague, and his team to quickly evaluate new combinations of enzymes on different types of bioenergy crops.

“ActE provides a natural example that is easy to grow in a test tube, and so provides many attractive starting points for further study,” Fox says. “As we begin to understand how it really works, we can then take that knowledge and improve on it.”

-M.B.

The opportunity to create renewable fuels from non-food crops is great, but first researchers need to break into the tightly-packed matrix of sugars that form plant cell walls. In this “Ask an Expert” video, Bruce Dale describes how GLBRC researchers use ammonia-based pretreatments to provide easier access to these sugars.

You can view more of our “Ask an Expert“ videos and ask your own questions by visiting glbrc.org/askanexpert or using the QR code below.

Ask an Expert Home Bruce Dale Video

What is Pretreatment?

Goal: To design energy crops that produce higher yields, contain more energy and are built of easy-to-access sugars

Research Progress: • A technology called Zip Lignin,™ which makes it easy

to break apart—or unzip—the lignin in plant cell walls to release the cellulose within, could significantly reduce the energy requirements needed to process biomass and therefore cut costs. Researchers have located and isolated a gene that could make this concept possible, and are now testing it in poplar trees.

• Using cutting-edge DNA sequencing technologies and high-throughput analysis software, researchers have screened thousands of plants to identify the genes that affect such key biomass traits as increased yield and digestibility.

Goal: To engineer low-cost, feedstock-flexible processing schemes to unlock plant sugars

Research Progress: • Using AFEX,™ researchers have created an alternate

form of cellulose that is five times more susceptible to breakdown by enzymes. This discovery paves the way for additional improvements to cellulosic biofuel processing.

• After scouring natural environments and narrowing down the potential players, researchers have focused their attention on a novel microbe called Streptomyces sp. ActE, which is just as capable at breaking down cellulose as T. reesei, a fungus that is used heavily by industry as a source of biofuels enzymes.

• GLBRC scientists developed a robotics platform called GENPLAT that tests the ability of new enzymes and enzyme cocktails to break down biomass of all sorts into fermentable sugars. The system, which can process up to 96 samples at a time, is far more efficient than the normal testing method.

Building Better Bioenergy Crops

Reimagining Plant Processing

From Field to Fuel: A selection of GLBRC research highlights

10 Great Lakes Bioenergy Research Center

Goal: To develop fuel technologies that are economically, environmentally and socially sustainable

Research Progress: • Planting perennial biofuel crops such as switchgrass could

increase the number of beneficial insects on the landscape and reduce the need for insecticides, which Midwestern farmers now spray on an extra 3.5 million acres to combat an uptick in crop pests. The increase in pests is the result of the ongoing conversion of natural habitat to farmland planted in annual crops, a trend called “landscape simplification.”

• The carbon costs of converting Conservation Reserve Program (CRP) lands to corn and soybeans is high – even when using no-till cultivation practices. Growing grasses rather than corn or soybeans on the existing 30 million acres of CRP land could maintain climate, wildlife and other conservation benefits indefinitely while providing a valuable bioenergy feedstock.

• A study confirmed that in order for farmers to start growing dedicated bioenergy crops, their net earnings from biomass crops would need to meet or exceed those from conventional crops and include a risk premium to account for the transition.

Goal: To create efficient and scalable ways —whether biological or chemical—to turn plant sugars into biofuels

Research Progress: • By combining gene expression and metabolism data with

in vivo testing, researchers have developed computational models that accurately predict how metabolism will change in response to particular mutations. This allows rational design of new microbes with improved fuel synthesis properties.

• GLBRC experts have increased yeast’s appetite for xylose, which is the second most abundant sugar in plant biomass. By encouraging these microbial powerhouses to consume a larger share of the available plant sugars, researchers could significantly increase the amount and speed with which biomass can be used to produce biofuels.

Creating Sustainable Landscapes

Converting Plant Sugars into Fuels

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12 Great Lakes Bioenergy Research Center

Conversion

Fourth Stop: ConversionOnce biomass has been pretreated

and the sugars released, GLBRC scientists work with bugs like yeast and

E. coli to optimize the way they churn through sugars and ferment them to

produce fuels.

Even though S. cerevisiae, an industrial yeast that has been used by brewers for centuries, is great at chewing through glucose, it hasn’t had much of an appetite for the five-carbon sugars like xylose—the second most abundant plant sugar—that make up a good part of the plant cell wall. That is, until now.

“Strains of yeast that are currently used for biofuel production can only convert xylose to ethanol very slowly and inefficiently,” says Dana Wohlbach, UW-Madison genetics researcher and GLBRC postdoc. “But the more sugars a yeast can consume, the better, since more sugar consumption means more ethanol.”

Researchers have identified a species of

yeast found in bark beetles that is able to efficiently use xylose. After engineering that species’ xylose-friendly genes into an industrial yeast, researchers found that the industrial yeast, too, could use xylose along

Our experts are examining both biological and chemical approaches to fuel production, including fermentation of biomass-derived sugars and direct chemical synthesis of fuels from biomass. By exploring biological and chemical pathways in parallel, we are working to create a suite of technology options for converting cellulosic biomass into advanced fuels.

Brian Pfleger, a GLBRC project leader, is working on producing hydrocarbons that could work in today’s engines and fuel pipelines.

132011 Science Report

with other sugars—a development that could significantly increase the amount and speed with which biomass sugars can be converted to biofuels.

Although encouraging bacteria and yeast to act as miniature biofuel factories shows incredible promise, GLBRC is putting a few other bets on the table.

“Ethanol will probably continue to have a place in the automotive industry in the U.S. and around the world for decades,” says GLBRC Director Tim Donohue, “but it is never going to be an acceptable biofuel for diesels or aviation or the shipping industry.”

Donohue is eager to expand the Center’s suite of fuels so that if an airline or shipping company comes knocking, they’ll find options to help them meet ambitious industry goals for reducing petroleum use. (The airline industry, for example, has committed to achieving carbon-neutral growth by 2020, as stated by the International Air Transport Association.) These industries are demanding ready-to-use fuels that can be “dropped in” to existing infrastructure such as engines, gas tanks and pipelines.

Demonstrating a vote of confidence in the emerging biofuels industry this December, U.S. Navy Secretary Ray Mabus and U.S. Department of Agriculture Secretary Tom Vilsack announced that the Defense Logistics Agency (DLA) signed a contract to purchase 450,000 gallons of advanced drop-in biofuel, the single largest purchase of biofuel in government history.

While the Navy fleet alone uses more than 1.26 billion gallons of fuel each year, this biofuel purchase is significant because it accelerates the development and demonstration of a homegrown fuel source that can reduce America’s, and our military’s, dependence on foreign oil.

“This unprecedented fuel purchase demonstrates the Obama Administration’s

commitment to seeking energy security and energy independence by diversifying our energy supply,” stated Secretary Mabus.

To meet this massive call for drop-in fuels, researchers at the GLBRC are leveraging knowledge about E. coli bacteria by applying new synthetic biology approaches to generate the cellulosic biofuel technologies needed.

“In an ideal world, you could go to the gas station and fill up your car or truck with a biofuel that is indistinguishable from today’s petroleum derived fuels,” says Brian Pfleger, GLBRC project leader and UW-Madison assistant professor of chemical and biological engineering. “If we can successfully develop methods of producing fuels from cellulosic biomass, we can reduce the need for new fueling infrastructure and the problems associated with that enormous change.”

Pfleger’s team is working to produce hydrocarbons that can work in today’s engines and fuel pipelines. As part of their quest to create a “green diesel,” researchers are relying on modified bacteria to convert sugar into building blocks that can later be used to create a variety of new products, including fatty acids, oils or biodiesel.

“Oils are an ideal source for biofuels, because they can be used for multiple forms of fuel, including biodiesel,” says Steve Slater, GLBRC scientific programs

manager. “While ethanol cannot be used in heavy transportation diesel engines, oils can be modified for diesel and gasoline replacements.”

Using the same sugar building blocks that can be upgraded into oils for fuel, researchers also hope to extend the technology in the future to create chemicals, bioplastics and other commercial products.

-M.B., A.V.

Read more about Pfleger’s fatty acid-producing microbes and other GLBRC technologies at glbrc.org/technologies.

“If we can successfully develop methods of producing fuels from cellulosic biomass, we can reduce the need for new fueling infrastructure and the problems associated with that enormous change.”

Biofuels generated by GLBRC get a reality check courtesy of UW-Madison Mechanical Engineer David Rothamer and the UW–Madison Engine Lab, where ethanol and other fuel precursors can be burned in engines to measure data on emissions and fuel performance. Participating in our “Ask an Expert” video series, Rothamer describes the characteristics that make ethanol a promising fuel.

You can view more of our “Ask an Expert“ videos and ask your own questions by visiting glbrc.org/askanexpert or using the QR code below.

What is Ethanol?

Ask an Expert Home David Rothamer Video

Picture or 2-3 pictures

14 Great Lakes Bioenergy Research Center

Sustainability

Fifth Stop: Sustainability How can GLBRC researchers be sure that successful fuel production at the

lab bench can be scaled up to meet the needs of a state, a region or a country?

One way is to look at the fuel from every angle—counting the inputs related to growing, transporting and converting plant material into fuel. By using robust modeling software, GLBRC researchers are examining the feasibility of potential fuels or technologies not just for scalability, but also for sustainability.

Much more than a buzzword, “sustainable” means that trade-offs—social, environmental and economic factors—have been measured, modeled and validated against actual “boots on the ground data” measured at agricultural research stations and on Midwestern farms, says Randy Jackson, a grassland ecologist and UW-Madison professor of agronomy who co-leads the Center’s sustainability research.

GLBRC research on bioenergy cropping systems, for example, has shown that such crops lead to everything from a reduced need for insecticide (due to an increase in beneficial insects) to increased bird and grassland diversity. “It’s really exciting that these systems offer the opportunity to actually improve both landscape

management and ecosystem services, or benefits, that we get from the land,” says Jackson.

For the Center, one such opportunity is the chance to provide research-based recommendations on the most suitable places to grow biofuels crops—an issue that has been the subject of intense debate.

According to a study conducted by GLBRC researchers at Michigan State University, farmers and policymakers should wait before converting Conservation Reserve Program (CRP) land to corn and soybean production.

This study, published in the Proceedings of the National Academy of Sciences in August 2011, shows directly for the first time that the carbon costs of converting CRP lands to corn and soybeans is high – even when care is taken to protect soil carbon from loss by using no-till cultivation practices.

Carbon debt results from carbon dioxide and other greenhouse gases released when land is converted from natural vegetation to agriculture. It’s called debt because until a

GLBRC Sustainability researchers have identified several key considerations in the choice of biofuel crop systems the are likely to lead to more sustainable outcomes, and they’ve dubbed them the “three Ps.” First, a successful biofuel crop must be productive for it to be economically profitable. Second, perennial crops are preferred for their ability to sequester carbon, reduce soil erosion and conserve biodiversity. Third is polyculture-tolerant, or the ability of potential biofuel crops to be grown in close proximity to other species or within short-term rotations.

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new biofuel crop creates enough renewable fuel to offset the lost CO2, the new biofuel crop has no climate benefit. In fact, it’s the same as burning fossil fuel as far as the atmosphere is concerned, says GLBRC’s Ilya Gelfand, an MSU postdoctoral researcher.

“Conversion creates carbon debt, which must be paid off before the biofuel crop can provide climate mitigation benefits,” Gelfand says. “No-till practices (planting without plowing) reduced by two-thirds the amount of debt created by the conversion, but still it would take 29 to 40 years for it to be repaid by growing corn and soybean for biofuel.”

Alternatively, growing CRP grasses harvested for cellulosic ethanol would create no debt and provide immediate energy and climate mitigation benefits, he added.

Nationally, more than 30 million acres are set aside as CRP land, and they provide significant climate, wildlife and other conservation benefits, says Phil Robertson, a co-author and MSU professor of crop and soil sciences who leads GLBRC sustainability research.

“Growing CRP grasses rather than using the land for corn or corn-soybean production could maintain these benefits indefinitely while providing a valuable bioenergy feedstock,” Robertson says. “It could be a win-win for farmers and the environment once a market for cellulosic biofuel develops.”

Another way to keep a cap on carbon is to minimize the number of miles that biomass must be transported to the factory gate.

In fact, in order to be economically viable, plant biomass would need to come from an area no more than 50 miles away from where it will be processed into fuel.

A GLBRC team is working to solve biofuel supply chain issues by exploring a new intermediate structure for gathering, processing and converting biomass into cellulosic biofuels. The proposed structures, called Regional Biomass Processing Depots (RBPDs), could improve both the economics and the sustainability of the biomass supply chain by keeping a portion of pretreatment and processing closer to the farm.

“By staging part of the processing closer to where biomass is grown, a distributed model like the RBPDs could help create jobs in rural communities while also transforming plant biomass into an energy intermediate that’s easier to transport,” says Pragnya Eranki, an MSU doctoral student in chemical engineering.

At the heart of the supply chain challenge is transportation.

“For cellulosic ethanol, we really don’t know what will happen between the field and the biorefinery,” says Bryan Bals, an MSU postdoctoral researcher. “Would it be stored on the farm in bales, sent to a storage facility or sent immediately to the biorefinery? Nobody really has that figured out yet.”

As modeled by the MSU research team, the RBPD concept looks somewhat like a web. Regional depots would be scattered across the state to perform pretreatment,

making the biomass more amenable to fuel conversion and compacting it for easier transport to a central biorefinery.

A recent life cycle assessment study conducted by Eranki and Bruce Dale, an MSU professor of chemical engineering, shows that this form of distributed network generates nearly the same net energy as the centralized biorefinery model, but generates significantly lower greenhouse gas emissions.

“If you can create a network of these depots across the country that can densify the biomass, you can ship it across the country without a major increase in cost for transportation,” says Bals.

-M.B.

At the Kellogg Biological Station in Hickory Corners, Mich., researchers transplant switchgrass from the greenhouse to experimental field plots.

The particular mixture of plant and animal species within a field or farm can have an impact on overall landscape health, which is an important factor in designing sustainable biofuels scenarios. As part of our “Ask an Expert” video series, Doug Landis talks about biodiversity and how it relates to biofuels research.

You can view more of our “Ask an Expert“ videos and ask your own questions by visiting glbrc.org/askanexpert or using the QR code below.

What is Biodiversity?

Doug Landis VideoAsk an Expert Home

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16 Great Lakes Bioenergy Research Center

Technology Transfer

The GLBRC is constantly inventing new technologies for improving the transformation of plant biomass into sustainable fuels. By leveraging our member institutions’ technology transfer and de-risking capacities, GLBRC researchers strive to produce, develop, and deliver fundamental biofuels technology breakthroughs. Wherever possible, the Center is interested in establishing research collaborations with private sector partners to enhance the flow of commercializable technologies from the lab to industry.  

Invention or Technology

Technology Transfer & Commercialization

Basic and Relevant Research

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Expected IDR production for $25.4 million/year at a major academic research institution

Invention Disclosure Reports (IDR)

Patent Applications

While the Center is focused on providing basic science breakthroughs that will form the foundation of the new bioeconomy, researchers also work with patent and licensing experts at the Wisconsin Alumni Research Foundation (WARF) and at MSU Technologies to disclose and license new research technologies that result from Center funding. In 2012, GLBRC inventions (shown above as Invention Disclosure Reports) surpassed the national average of expected disclosures for $25.4 million of funding each year at a major academic research institution.

172011 Science Report

To learn more about GLBRC Technologies, visit www.glbrc.org/industry/technologies

Moving Research from Bench to Biorefinery

No technology starts out at a commercially relevant scale. Before companies or entrepreneurs are willing to invest in an emerging technology, they want to see the proof of concept and probably some scale-up work.

Making cellulosic biofuels requires several steps. First, the biomass must be grown and harvested. Next, the biomass requires pretreatment and processing into a usable fuel.

Pretreatment of biomass is the most expensive step in the biofuels conversion process because not all plants respond the same way to the same pretreatments. AFEX,™1 a pretreatment process that combines ammonia and pressure, is expected to enable the rapid deployment of advanced biofuels because it breaks open plant cell walls and makes plant materials more easily accessible. At his Michigan State University lab, Bruce Dale and his colleagues have been working to make AFEX™ as efficient and productive as possible.

For ammonia fiber expansion technology, or AFEX,™ the research began with equipment the size of today’s big screen televisions, which had the capacity to produce five gallons of pretreated biomass. So, how has this technology grown to the expectation of processing one ton of biomass per day?

“We have known for some years that AFEX™ is a very effective pretreatment on the lab scale,” says Dale, GLBRC Deconstruction leader and MSU professor of chemical engineering and materials science. “But, we had not been able to figure out a low cost way of applying it on a large commercial scale.”

To bridge this gap between lab-scale basic research and commercial-scale applications, GLBRC has partnered with MBI, a subsidiary of Michigan State University’s Foundation.

“Our collaboration with the engineers at MBI has helped us develop a low-cost version of AFEX,™ which is now being scaled up for commercial applications,” Dale says. “It’s been a very successful story.”

Their close association with both industry and universities helps give MBI the perspective to inform research priorities

and advance technology toward commercialization.

“As a non-profit organization, we can handle intellectual property in a noncompetitive way. You can’t get that when working with a company,” says MBI President and CEO Bobby Bringi.

“The basic research going on at the Center has the potential to improve advanced biofuels at many points along the pipeline,” says Dave Pluymers, GLBRC’s intellectual property manager. “MBI takes this basic research and assesses its commercial potential.”

Though promising, an emerging technology like AFEX™ is not without risk. Actionable de-risking roadmaps are part of the service MBI provides the GLBRC. It’s a systematic way to articulate and label the risks involved with an early-stage technology and then address them to increase commercial potential.

“MBI will scale up technology incrementally from the lab bench to a commercial-sized application in an attempt to prove the technology’s value and appeal,” Pluymers says. “For this reason, de-risking is an important part of the process.”

“With Bruce Dale’s work, we needed to facilitate the AFEX™ process at a large scale, prove the concept and deal with the risks,” Bringi says. “MBI developed a novel reactor to carry out the technology. The breakthrough was getting it to the point of low-cost and high throughput. The new AFEX™ reactor is expected to be 30% less costly than prior reactor designs.”

This research and collaboration culminated in a U.S. Department of Energy grant of $4.3 million for the AFEX™ project. The grant will help scale the 10-liter lab reactor up to 1,000 liters and demonstrate AFEX™ treated biomass as a potential commercial feedstock.

“After de-risking the reactor, we de-risk the applications,” Bringi says. “We’re going to use the reactor to create significant quantities of biomass. Following which we

can demonstrate the value of AFEX™ to industry.”

Bringi expects the AFEX™-treated biomass to prove commercial significance by demonstrating that the process results in sugars that can be fermented to produce a wide range of bioproducts, and by confirming its use as an animal feed.

“After this, we’ll be much closer to commercial viability,” Bringi says. “Then we can take this technology and deploy it as widely as possible.”

-E.A.

View GLBRC’s Technologies on our website or on the Department of Energy’s Energy Innovation Portal. There you’ll find more information on each technology, including an overview of the invention, its benefits and potential applications. Go to glbrc.org/industry/technologies or follow one of the QR codes below to see more.

GLBRC TechnologiesDOE Energy

Innovation Portal

Technology

18 Great Lakes Bioenergy Research Center

Shaping Young Scientists, One Summer at a Time

When Jillian Foerster started her freshman year at Grand Valley State University in Allendale, Michigan, she knew that majoring in cell and molecular biology would open up several avenues for research exploration. What she didn’t know is that she would end up in Wisconsin studying biofuels.

“[With this major], you could do all these different things. You could go make wine. You could go into bioinformatics,” she says. “One of the things that interested me the most was renewable fuels. So, I just sat at the computer and Googled bioenergy internships.”

When she landed on the Research Experience for Undergraduates (REU) program offered by GLBRC, Foerster applied right away.

The REU programs, offered at UW–Madison and MSU, provide a cutting-edge research experience to undergraduate students interested in renewable energy. Each summer students participate in hands-on, contemporary bioenergy research with GLBRC scientists in the field or lab. They explore a wide range of topics including sustainability, biofuel production and considerations that are driving interest in

new bioenergy technologies. By experiencing the research

firsthand, conducting their own experiments, students are exposed to the process and culture of large-scale science.

“One of the biggest things I learned at the REU program is that scale of research and how it works,” Foerster says.

Gina Lewin, a summer 2009 REU student, is now a research assistant and graduate student working for the Center and continuing her education in microbiology.

“I went to a small undergrad school that had a lot of research, but nothing like they have here,” Lewin says. “For me the REU program was a really good chance to experience what research was like at a big institution and try to figure out if I was interested in grad school.”

Last summer, four students from around the country traveled to Madison to participate in the bioenergy REU. They spent ten weeks working with GLBRC mentors to complete cutting-edge research in the field.

In addition to providing a unique research experience, the REU program also emphasizes the ability to think critically.

As Foerster researched the very specific science of root architecture and lodging, she also participated in discussions with other students about broader topics, giving her the ability to think critically and talk about big picture science.

“I can say with confidence, that I would not be in grad school if I had not gone to this program,” Forester says.

Connecting Classrooms to Cutting-Edge Science

Students in Craig Kohn’s biotechnology classroom are on the hunt for biomass degraders. Scavenging for unique samples in piles of leaves or compost bins, they can use a simple filter paper test to determine if microbes within each environmental sample are capable of growing on the plant material.

Education and Outreach

The Center’s Education and Outreach group provides programs and resources to help broaden the understanding of current issues in bioenergy for the general public, and students and educators at the K-12, undergraduate and graduate levels.

“For me the REU program was a really good chance to experience what research was like at a big institution and try to figure out if I was interested in grad school.”

For more information on GLBRC’s opportunities for undergraduates, visit glbrc.org/education/programs

192011 Science Report

As they search, they’re doing more than completing an activity in their bioenergy unit — they’re searching for microbes that could play a role in creating biofuels more efficiently. In the process they’ll also get to experience scientific research that has not yet been described in their textbooks.

Since participating in the 2011 Bioenergy Institute offered by the GLBRC, Kohn, a science teacher at Waterford Union High School in Wisconsin, has been able to bring these real world research activities into his classroom.

During the week-long institute, Kohn says, “I got a really strong grasp of what bioenergy is and the realistic outlooks of what it could do, what it could be, and its limitations.”

The Bioenergy Institute is one of several programs GLBRC offers to assist educators with integrating bioenergy lessons into their curriculum. The programs bring researchers, teachers and curriculum coordinators together to form collaborations and produce high-quality educational materials.

Kohn took the knowledge he gained at the Institute and brought it back to his classroom, adding a biofuels component into a pre-existing biotechnology class. Using a combination of materials from the Center’s Education and Outreach team, Kohn provided students with an overview of biofuels within the context of biotechnology.

After completing the Bioenergy Institute, teachers have the option of gaining a more in-depth experience by participating in the Research Experience for Teachers (RET) program. As RET participants, teachers work very closely with specific GLBRC researchers to develop educational materials

that integrate bioenergy into lessons designed to teach fundamental scientific concepts.

In Kohn’s case, his participation in the summer 2011 RET program led to a collaboration with GLBRC‘s Cameron Currie, who explores symbiotic communities like those of the leaf-cutter ant. Currie’s research has taken him to locations in Central America in search of unique biomass-degrading microbes. Yet, when he gets samples back to the lab, one of the first tests his researchers use is the filter paper activity being used by Kohn’s students as they search for new biodegrading microbes.

With fresh activities, teachers have new tools at their disposal to interest their students in science.

“For a teacher to be able to spend a summer connected to a research lab and do some hands-on work really gives them a much better, more intuitive sense of what contemporary science is all about,” says John Greenler, GLBRC education and outreach director. “We want the teachers in the lab so they can get revved up about science and be able to communicate that excitement to their students.”

The Bioenergy Institute and the RET program work in tandem to provide teachers with an in-depth understanding of current bioenergy research and the tools necessary to bring that fundamental

knowledge into their classrooms.

The Educational Materials section of the GLBRC website features more than a dozen ready-to-use classroom activities ranging from bioprospecting to fermentation, several of which were designed by RET program participants.

“We’ve been thrilled to see such high demand for our activities,” says Greenler. “This level of

interest really shows us that teachers are looking for unique ways to teach science in their classroom. By using bioenergy as contemporary topic, students can get excited to work on problems that are still being solved by scientists.”

-A.V., M.B.

Craig Kohn runs through his new bioprospecting educational activity in a lab at the Microbial Science Building in Madison, Wisc.

GLBRC educational materials were developed by teachers and professional educators with input from our scientists. Many of the techniques described are the same, or closely mimic those conducted by researchers

within the Center, with adaptations made as necessary to work within the constraints of the K-16 classroom. You can view all of our educational materials by visiting glbrc.org/education or using the QR code.

Educational Materials

Great Lakes Bioenergy Research Center - w w w.glbrc .org/education

AAAS. 1993. Benchmarks for Science Literacy: Project 2061. Oxford University Press. New York

1T

Field Investigations: Biomass Yield and Carbon Cycling in Crops

Levels6 through undergraduate

SubjectsScience, Environmental Studies

Objectives• Design, conduct and analyze

an experiment to measure primary productivity and/or soil respiration rates in a field.

• Describe the movement of carbon through a field ecosystem, including above and below-ground components.

• Explain how land management practices (tilling, fertilization, etc) and different plants (prairie, grass, etc) have an effect on carbon cycling.

Materials• Field Investigations: Biomass

Yield and Carbon Cycling in Crops Package

Activity TimeHighly variable: Two 50-minute classroom period minimum plus wait time for plant growth.

Standards1B The Nature of Science:

Scientific Inquiry5E The Living Environment: Flow

of Matter and Energy8A The Designed World:

Agriculture11A Systems

Overview: Field investigations to strengthen student understanding of the ability of plants to sequester carbon above and below ground, and the role soil and soil microbes play in the carbon cycle. Students will measure primary productivity above and below ground, and infer cellular respiration rates of soil microbes in grassland environments. These activities are adaptable to school-yard plots, existing agricultural plots or natural areas.

beta version

Educational Materials

DResearch CentersOE Bioenergy GLBRC.org

Copyright © 2012. All rights reserved.

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Technology

David Pluymersdpluymers@glbrc.wisc.edu

Operations

Dan Laufferdlauffer@glbrc.wisc.edu

Media

Margaret Broerenmbroeren@glbrc.wisc.edu

Contact Us

Education & Outreach

John Greenlerjgreenler@glbrc.wisc.edu

Science

Ken Keegstrakeegstra@msu.eduSteve Slaterscslater@glbrc.wisc.edu

Credits

Published by the U.S. Department of Energy’s Great Lakes Bioenergy Research Center (GLBRC)Produced by the GLBRC Communications DepartmentWriters: Margaret Broeren, Eric Anderson, Amanda VoyeDesign and Photography (unless otherwise noted): Matthew Wisniewski

NewsletterGLBRC.org/newsletter