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Instruments' role in the biofuels roadmap.

One only needs to read the paper or watch the news to realize that fossil fuel has a tenuous future. The reserves are finite and cannot be replenished. Many of the regions where oil comes from are shaken by political turmoil. Furthermore, the impact these fuels have on the environment is considerable. There is no easy solution to figs problem, but one potential answer lies with biofuels.

According to Project Finance International, investments in the biofuels industry totaled $2.24 billion between 2000 to June 2006. The global market for biofuels in 2005 grew 15% from 2004 to $15.7 billion, and estimates by Clean Edge forecast that the market will reach $52.5 billion by 2015. Since mid-2005, $1 billion has been spent on building new ethanol plants. Investments ill biofuels are being made by both the public and private sector. Analytical instrumentation will play a vital role in making biofuels a reality. In addition, as a recent US project indicates, biofuel research will require developments in instrumentation that, if realized, could advance instrument technology. In June of this year, BP announced that it would supply $500 million over the next ten years to build a dedicated biosciences energy research lab. France established an ambitious biofuels plan, with usage goals of 5.75% by 2008, 7% by 2010, and 10% by 2015.

In his 2006 State of the Union Address, President Bush announced the Advanced Energy Initiative, which aims to limit the United States' dependence on foreign oil. The Initiative includes $2.1 billion in funding for the Department of Energy (DOE) for more research to be done on alternative sources of energy, including biofuels. The president's 2007 budget requests $1.2 billion for the DOE's office of Energy Efficiency and Renewable Energy for research and development of alternative fuel sources. The DOE has announced a goal of replacing 30% of transportation fuel with biofuels by 2030.

In June the "Breaking the Biological Barriers to Cellulosic Ethanol: A Joint Research Agenda" white paper was published, providing a scientific roadmap for the conversion of biomass to biofuels. A result of a 2005 workshop organized by the DOE's Office of Science and the Office of Energy Efficiency and Renewable Energy, the white paper examines the potential application of new technologies and instrumentation to the creation of biofuels.

The transformation of feedstock biomass to ethanol is not easy. Ethanol is a byproduct of the fermentation of sugars and can be obtained from a variety of feedstocks, such as corn, the United States' prime source, as well as from switchgrass, poplar trees and sugarcane, which has been a very successful source of biofuels for Brazil. The most significant obstacle to extracting ethanol from plants is the feedstock's recalcitrance to being processed as ethanol.

Lignin, which is bonded to the cellulose in the cell walls of a feedstock, provides a plant with structural stability and does not break down easily. Yet cellulose must be broken down into glucose, which is then fermented into ethanol. One answer to this problem of breaking down lignin is manipulating the genetic structures of the feedstock in order to grow plants with lower lignin content. Research is also being done into the formation of lignin and how it connects to other parts of the plants' cell walls.

Breaking cellulose and hemicellulose down into their constituent sugars is another challenge. To do so, enzymes are needed that will convert the polysaccharides once they are separated from the other components of the cell wall. Due to the heterogeneous nature of the cellular environment, discovering functional enzymes is a complicated process.

Once the sugars are obtained, they need to be fermented into ethanol. This requires finding microbes that can survive in sugar- and ethanol-rich environments and perform the fermentation process. Genetic study of the microbes will yield information that will help researchers develop microbes that are more efficient producers of ethanol.

The roadmap foresees three phases. The research phase, five years into the project, will focus on understanding the feedstock from a cellular perspective, optimizing enzymes that will break biomass down into polysaccharides, and improving the fermentation process. The technology deployment phase, 10 years into the project, will consist of growing feedstock that is optimal for being converted to ethanol and combining the breakdown and fermentation processes into a single step. After 15 years, the systems-integration phase will make further use of these optimal feedstocks in biorefineries, where the process of breakdown and fermentation will be further accelerated to become economically viable.

The white paper specifically notes what analytical techniques will be needed and how recent developments in systems biology and specifically transcriptomics, proteomics, glycomics, lignomics, and fluxomics will be applied to the problems of biomass conversion. Among the specific techniques highlighted in the white paper are nuclear magnetic resonance, mass spectrometry, gas and liquid chromatography, electron microscopy, atomic force microscopy and microarrays. However, as James Frederickson, a participant in the workshop that produced the white paper explains, "There's no one single magic bullet for these [barriers to converting feedstock into ethanol], so I think it's very important to be able to take these multiple technologies and approaches that can provide complementary information to try and resolve some of these issues."

Nuclear magnetic resonance spectroscopy (NMR) will be used to characterize and analyze plant structures. Dr. Frederickson said nuclear magnetic resonance spectroscopy (NMR) was "a major contribution to analyzing the structure of cell walls and that kind of information and detail has lacked research to this point in time." Specifically, NMR will be used to create 2D and 3D maps of cell wall composition. "NMR can also be useful to characterize the nature of breakdown products as well as the residual material," he added. NMR will also be used to complement certain mass spectrometric applications, such as characterizing metabolites, analyzing lignin, and monitoring the process of polysaccharide hydrolysis, and will be used to study metabolic pathways. Dr. Frederickson pointed out that one area of NMR technology that could be improved was its spatial resolution.

Mass spectrometry (MS) will be used as part of the project to characterize metabolites, analyze lignin and monitor the process of polysaccharide hydrolysis. "Separations and then mass spectrometry will end up being essential for a lot of these applications," explained Dr. Frederickson. In recommending improvements, Dr. Frederickson alluded to the complex mix of proteins in the samples being studied and said that the resolution of the technique could be increased.

In addition to its use in tandem with MS, liquid and gas chromatography will also be used to analyze the components of plant cell walls. According to Dr. Frederickson, chromatography is also important to be able to characterize cellular metabolites to determine how energy and materials are flowing through the cell. Because these metabolites have very short half-lives, there is always a need for faster separations, he noted.

Being able to create images of cellular structures will also be an important part of biofuels research. Referring to the utility of variants of electron microscopy in the project to come, Dr. Frederickson commented that "some of the systems now that are using the cryo-EM [cryoelectron microscopy] are actually getting quite good at characterizing the structure of even relatively large proteins and other types of molecular complexes. And so, I think there's potentially some complementarity between some of those electron microscopy techniques and the NMR." In addition, the increasing capacity of scanning electron microscopy to analyze samples that contain moisture will be utilized as part of the project. Electron microscopy can also be used to characterize cellular structures.

Dr. Frederickson was particularly optimistic about the use of microarrays in the project. "Arrays are very good in terms of being able to, using a relatively high throughput approach, expose cells to a variety of conditions and then you can use various types of modeling approaches to start to pull out those regulatory networks based on this expression analysis." When asked what kinds of improvements could be made on the technology, Dr. Frederickson replied, "I think the improvements have more to do with the design of the array and increasing the content in terms of coverage of the genome and the intragenic region," said Dr. Frederickson. The white paper calls for more microarrays that are specifically targeted to new plant varieties and microorganisms, and, given the heterogeneous populations of microbes that will be studied, for single-cell gene-expression methods applicable to diverse cell types.

Atomic force microscopy (AFM) is another technique discussed in the white paper. One of the advantages of using AFM is that the technique can be used in a liquid environment. "One of the major limiting factors is obviously the aspect of having the appropriate catalytic enzymes that are able to engage with and function on relatively heterogeneously comized." AFM will enable the observation of biomass interacting with various enzymes.

The road to a thriving biofuels program is not without its complications and frustrations. Given our situation, however, it is the right road to travel, and analytical instruments should help lead the way.
Advanced Energy Initiative 2006 Funding for Energy
Efficiency and Renewable Energy Programs

Hydrogen Technology 13%
Fuel Cell Technology 12%
Vehicle Technology 29%
Biomass 14%
Solar 13%
Wind 6%
Geothermal 4%
Program Management 9%

Note: Table made from pie chart.
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Publication:Instrument Business Outlook
Date:Sep 30, 2006
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