Progress 10/01/02 to 09/30/07
OUTPUTS: Canola planting trial. MSU has just begun a study to grow 200 acres of canola in the 2007-08 growing season. The goal is to determine costs, yields, losses, and potential challenges with canola as an energy crop for Michigan. Life-cycle Assessment. MSU has assessed various parameters to improve the overall environmental performance of corn based ethanol & soy biodiesel production. Optimization of ethanol fermentation. We are working to understand how various pretreatmens and enzymes act synergistically to hydrolyze cellulose and hemicellulose to fermentable sugars. We are determining how pretreatment conditions affect various species that might build up in process recycle streams. We are determining how best to modify plant cell wall composition for optimum response to pretreatment and enzymatic hydrolysis. We are working to reduce the overall cost of the ammonia fiber expansion (AFEX) process by careful analysis of overall process economics. Together with Purdue university
we have demonstrated that pretreated poplar hydrolyzate can be fermented to ethanol with 93% efficiency. We have been working on dry distillers grains (DDG). We have optimized the AFEX pretreatment and have demonstrated the fermentability of sugar to higher ethanol yield. In addition we have shown the amino acid balance and feed value on enhanced DDG. Biodiesel process development. MSU is developing reactive distillation as a novel, energy efficient, low capital cost route to biorenewable fuels and chemicals. MSU is working to develop a continuous process that produces biodiesel and utilizes the glycerol byproduct simultaneously. Biodiesel fuel composition and properties. MSU is investigating the effect of composition of fuel on the properties of fuel with primary emphasis on improvements of the cold flow properties by altering composition. Ozonolysis for biodiesel modification. Studies continue on the use of methanol-mediated ozonolysis for modification of soy methyl esters. Corncob
granules for biomaterials. Corncob granules have been obtained using an AFEX and silane treatments and evaluated for biocomposites. Biosensors and Biocatalysts. MSU has developed nanostructured bioelectronic and biomimetic interfaces containing proteins incorporated into interfaces containing either polyelectrolyte multilayers or tethered lipid bilayers. Reactive Distillation for Specialty Chemicals. In this work we are focusing on potential routes to utilize streams from fermentation streams producing succinic acid, considering products including esters. Properties of bio-derived compounds. We are developing a knowledge base of physical and thermodynamic properties for biorenewable chemicals. Many of the properties are not available in the literature. We perform measurements and predictions. MSU is offering a course entitled "Green and Sustainable Engineering" for the fourth time this Spring, 2008 semester. Michigan State University is developing a Biorefinery Training Facility,
funded by the Department of Labor, that will train Michigan workers to participate in the bioeconomy.
PARTICIPANTS: Kris A. Berglund Bruce E. Dale Carl T. Lira Dennis J. Miller Ramani Narayan R. Mark Worden
TARGET AUDIENCES: Cross-State Facilitation
PROJECT MODIFICATIONS: None.
This project has fostered important communication between universities to provide a change in knowledge for all parties involved. Further, this project has further enhanced all investigator's ability to reflect the state of the art when disseminating information through publications.
- Hassler, B. L.; Dennis, M.; Laivenieks, M.; Zeikus, J. G.; Worden, R. M., Mutation of Tyr-218 to Phe in Thermoanaerobacter ethanolicus Secondary Alcohol Dehydrogenase: Effects on Bioelectronic Interface Performance. Applied Biochemistry and Biotechnology 2007, in press.
- Kohli, N.; Hassler, B. L.; Parthasarathy, L.; Richardson, R. J.; Ofoli, R. Y.; Worden, R. M.; Lee, I., Tethered lipid bilayers on electrolessly deposited gold for bioelectronic applications. Biomacromolecules 2006, 7, (12), 3327-3335.
- Hassler, B. L.; Kohli, N.; Zeikus, J. G.; Lee, I.; Worden, R. M., Renewable dehydrogenase-based interfaces for bioelectronic applications. Langmuir 2007, 23, (13), 7127-7133.
- Hassler, B. L.; Worden, R. M., Versatile bioelectronic interfaces based on heterotrifunctional linking molecules. Biosensors & Bioelectronics 2006, 21, (11), 2146-2154.
- Asthana, N.S.; Kolah, A.K.; Vu, D.T.; Lira, C.T.; Miller, D.J. A Kinetic Model for Esterification of Lactic Acid and Its Oligomers, Ind. Eng. Chem. Res. 45, 5251-5257 (2006).
- Bals, B., Dale, B. E. and Balan, V. Enzymatic Hydrolysis of Distiller's Dry Grains and Solubles (DDGS) using Ammonia Fiber Expansion Pretreatment Energy and Fuels 20, 2732-2736, (2006).
- Laureano-Perez, L., Dale, B. E., O'Dwyer, J. P. and Holtzapple, M. Statistical Correlation of Spectroscopic Analysis and Enzymatic Hydrolysis of Poplar Samples Biotechnology Progress 22, 835-841, (2006).
- Shishir P. Chundawat, Balan Venkatesh and Bruce E. Dale. Effect of Corn Stover Composition and Particle Size on AFEX Pretreatment and Enzymatic Hydrolysis Biotechnology and Bioengineering 96, 213-219, (2006)
- Tamika Bradsha, Hassan Alizadeh, Farzaneh Teymouri, Balan Venkatesh and Bruce E. Dale. Ammonia Fiber Expansion Pretreatment and Enzymatic Hydrolysis of two growth stages of Reed Canary Grass Applied Biotechnology and Bioengineering. Vol. 136-140, 395-406. (2007)
- Callista Ransom, Venkatesh Balan, Gadab Biswas, Bruce E. Dale, Elaine Crockett and Mariam Sticklen. Heterologous Acidothermus cellulolyticus 1,4-β-endoglucanase E1 Produced with in corn biomass converts corn stover into glucose Applied Biotechnology and Bioengineering. Vol. 136-140, 207-220. (2007)
- Hesham Oraby, Balan Venkatesh, Bruce E. Dale, Rashid Ahmad, Callista Ransom, James Oehmke and Mariam Sticklen. Enzyme for Biofuel; High-Level Production of endo-1, 4-b-glucanase in Transgenic Rice with Polysaccharides into Fermentable sugars Transgenic Research. (2007)
- Masud Huda, Balan Venkatesh, Lawrence Drzal, Shishir P. Chundawat, Bruce E. Dale and Manjusri Misra. Effect of Ammonia Fiber Expansion (AFEX) and Silane Treatments of Corncob Granules on the Properties of Renewable Resource Based Biocomposites J. of Biobased Materials and Bioenergy. 1, 127-136 (2007).
- Kohli, N.; Vaidya, S.; Ofoli, R. Y.; Worden, R. M.; Lee, I., Arrays of lipid bilayers and liposomes on patterned polyelectrolyte templates. Journal of Colloid and Interface Science 2006, 301, (2), 461-469.
- Kohli, N.; Worden, R. M.; Lee, I., Direct transfer of preformed patterned bio-nanocomposite films on polyelectrolyte multilayer templates. Macromolecular Bioscience 2007, 7, (6), 789-797.
- Vu, D.T.; Lira, C.T.; Asthana, N,S; Kolah, A.K. Miller D.J. Vapor-liquid equilibria in the systems Ethyl Lactate + Ethanol and Ethyl Lactate + Water, J. Chem. Eng. Data. 51, 1220-1225, (2006).
- Kolah, A.K.; Asthana, N.S.; Vu, D.T.; Lira, C.T.; Miller, D.J. Reaction Kinetics of the Catalytic Esterification of Citric Acid with Ethanol, Ind.Eng. Chem. Res., 46, 3180-3187 (2007).
Progress 01/01/06 to 12/31/06
The elevated pressure reactive distillation column designed by Dr. Lira and Dr. Miller is in its final stages of construction. All column components have been acquired and are being installed. The supporting framework for the column is in place. The column is expected to be operational in February, 2007 and will be immediately put to use for two new projects that Dr.Miller and Dr. Lira and leading. One project, funded by the MEDC 21st Century Jobs Fund, is the production of succinic acid esters via reactive distillation. The second project, funded by the USDA NRI Program, is for the use of reactive distillation to produce an improved composition of biodiesel. The successful proposals submitted for these projects were significantly strengthened by the commitment of funds from the USDA 'Enabling Biorefineries' project. Mark Worden's lab is using nanofabrication methods to develop new, high-performance catalytic interfaces containing dehydrogenase enzymes, which carry
out oxidation and reduction reactions. The resulting bioelectronic interfaces can use spontaneous chemical reactions to generate electricity (biological fuel cells) or use electricity to drive chemical reactions (biocatalytic reactors). Site-directed mutagenesis resulted in production of a new secondary alcohol dehydrogenase enzyme that used NADH as opposed to the much more expensive NADPH. Also, methods have been developed that allow the labile components of the bioelectronic interfaces to be regenerated. Dr. Bruce Dale's lab is working on developing integrated pretreatment, hydrolysis and fermentation systems to convert cellulosic biomass to fuels such as ethanol. Five doctoral students and a post doc are currently involved in these integrated projects. Among the important advances achieved this year are: reduction in the amount of ammonia required and concentration of ammonia required for effective application of the ammonia fiber expansion (AFEX) process, better understanding of
the fundamental mechanisms that make AFEX effective, development of microplate screening methods for high throughput analysis of treated cellulosic biomass and the first demonstration that pretreated biomass (by AFEX) can be fermented by recombinant organisms without any detoxification or nutrient supplementation. Detoxification and nutrient supplementation are absolutely required for acid pretreated biomass. A major corn dry miller is now involved in scaling up and commercializing AFEX. The 2006 multi-state meeting was attended by Professor Dennis Miller.
The reactive distillation work will facilitate the simultaneous production and purification of biobased products, thereby making them more competitive with petroleum derived products. Biodiesel economics will be improved by this process. The large scale column will give us the opportunity to test concepts and develop process economics in a way that cannot be done with smaller devices. Biological fuel cell technology is improved by the nanofabrication technology. This marriage of biotechnology and nanotechnology is among the most fertile areas for advancing the use of biology to process agricultural materials for chemicals and fuel production. Finally, the work on cellulosic ethanol production is directly related to President Bush's call to reduce the cost of ethanol production from domestic energy crops such as switchgrass. The intense interest in this area is highlighted by the fact that the AFEX process is now in the early stages of commercial application.
- Vu, D.T.; Lira, C.T.; Asthana, N,S; Kolah, A.K. Miller D.J. Vapor-liquid equilibria in the systems Ethyl Lactate + Ethanol and Ethyl Lactate + Water, J. Chem. Eng. Data. 51, 1220-1225, (2006)
- Brian L. Hassler, Megan Dennis, Maris Laivenieks, J. Gregory Zeikus, and Robert M. Worden (2007) Mutation of Tyr-218 to Phe in Thermoanaerobacter ethanolicus secondary alcohol dehydrogenase: effects on bioelectronic interface performance, Applied Biochemistry and Biotechnology, (in press)
- Dale, B. E. Impacts of Ethanol on the Farm Economy. A High Growth Strategy for Ethanol. p. 31-42. Report of an Aspen Institute Policy Dialogue, published by the Aspen Institute, Queenstown, Maryland (2006)
- Bals, B.; Dale, B. E.; Balan, V. Enzymatic Hydrolysis of Distillers Dry Grains and Solubles (DDGS) using Ammonia Fiber Expansion Pretreatment. Energy and Fuels 20, 2732-2736, (2006).
- Saffron, C.; M., Park, J. H.; Dale, B. E.; Voice, T. C. Kinetics of Contaminant Desorption from Soil: Comparison of Model Formulations using the Akaike Information Criterion. Environmental Science and Technology 40, 7662-7667, (2006).
- Laureano-Perez, L.; Dale, B. E.; ODwyer, J. P.; Holtzapple, M. Statistical Correlation of Spectroscopic Analysis and Enzymatic Hydrolysis of Poplar Samples. Biotechnology Progress 22, 835-841, (2006).
Progress 01/01/05 to 12/31/05
Mark Worden's lab is using nanofabrication methods to develop new, high-performance catalytic interfaces containing dehydrogenase enzymes, which carry out oxidation and reduction reactions. The resulting bioelectronic interfaces can use spontaneous chemical reactions to generate electricity (biological fuel cells) or use electricity to drive chemical reactions (biocatalytic reactors). A novel approach that uses the amino acid cysteine to couple dehydrogenase enzymes, their cofactors, and electron mediators to an electrode has been developed and analyzed using cyclic voltammetry and chronoamperometry. The approach has been shown to work effectively for a variety of dehydrogenase enzymes, including sorbitol dehydrogenase, alcohol dehydrogenase, and secondary alcohol dehydrogenase. Over the past several years, Dennis Miller's group has collaborated with James Jackson in the Department of Chemistry at MSU to develop active catalysts and reaction conditions that carry out
several reactions that are key to the future biorefinery. Among these are the hydrogenation of carboxylic acids to the corresponding alcohol with retention of acid stereochemistry in the alcohol, condensation chemistry to form highly-functionalized monomers, and selective carbohydrate hydrogenolysis to convert biomass sugars to high-volume diols and polyols. Bruce Dale's lab is optimizing the ammonia fiber explosion (AFEX) process, which treats lignocellulosic biomass with liquid ammonia under pressure followed by explosive pressure release to enhance conversion of structural carbohydrates (cellulose and hemicellulose) to fermentable sugars. AFEX treatment increases the susceptibility of lignocellulose to enzymatic hydrolysis by decrystallizing cellulose, prehydrolyzing hemicellulose, reducing lignin content of the treated material, and greatly disrupting the fibrous structure of biomass. Theoretical yields of glucose from AFEX treated corn stover have been achieved at modest enzyme
loadings. Current research focuses on achieving theoretical xylose yields. Dr. Dale's lab is using life cycle analysis (LCA) to assess the environmental impacts of products and processes and thereby improve their sustainability. We have learned that it is possible to remove very large fractions of corn stover for conversion to fuels and chemicals if appropriate management tools are used, such as no till agriculture and cover crops. Surprisingly, residue removal reduces soluble nitrogen leaching from corn fields. Reactive distillation to simultaneously form and purify biobased fuels and chemicals Collaborative research involving Dennis Miller and Carl Lira is applying reactive distillation to produce chemicals in which the conversion reactions are equilibrium limited, including the recovery of diols (produced from biomass sugars) from aqueous solution via formation of their acetals, which have boiling points below that of water and are thus easily separated, the formation of organic
acid esters, particularly ethyl lactate, from fermentation-derived feedstocks, and the transesterification of plant triglycerides to form alkyl esters, e.g. biodiesel.
The nanostructured interfaces containing dehydrogenase enzymes have been used to develop novel biosensors and biosensor arrays and are now being adapted to design electrobioreactors suitable to produce biobased products. Products formed using the catalysts recently developed have potential to replace current petroleum-based monomers, solvents, and other products, as they can have improved function and lower cost as well as being renewable. In addition, reactive distillation should have a significant impact on biorefinery efficiency, as simultaneous reaction and purification in a single piece of process equipment substantially reduces capital and operating costs relative to conventional processing. A major roadblock to ethanol fuel production is the inherent resistance of lignocellulosic materials toward conversion to fermentable sugars. The AFEX pretreatment research is making significant improvements in the efficiency and economy of pretreatment, thus contributing to
the economic outlook for fuel ethanol. For the first time, we can design a new industry so that it meets both economic and environmental criteria. Recent life-cycle analysis research results that include analysis of ethanol's net energy debunk the notion that a negative net energy has any value, particularly to guide public policy. This result clarifies the need for commercial processes to produce biobased fuels and chemicals.
- Hassler, B. and R.M. Worden, "Versatile bioelectronic interfaces based on heterotrifunctional linking molecules," Biosensors and Bioelectronics, Online 11 November (2005).
- Kolah, A.; Lira, C.T.; Vu, D.T.; Asthana, N; Miller, D.J. "Modeling of oligimerization in concentrated lactic acid solutions", Fluid Phase Equil. 236, 125-135 (2005).
- Asthana, N.; Kolah, A.; Vu, D. T.; Lira, C.T. ; Miller D.J. "A Continuous Reactive Separation Process for Ethyl Lactate Formation", J. Org. Process Res. Dev. 9, 599-607, (2005).
- Kim, S and B.E. Dale, (2005) "Ethanol Fuels: E10 or E85 - Life Cycle Perspectives," The International Journal of Life Cycle Assessment, (Online First, 1-5).
- Kim, S and B.E. Dale, "Life Cycle Assessment Study of Biopolymers (Polyhydroxyalkanoates) Derived from No-Tilled Corn," The International Journal of Life Cycle Assessment, 10 (3) 200-210 (2005).
- Kim, S and B.E. Dale, "Environmental aspects of ethanol derived from no-tilled corn grain:nonrenewable energy consumption and greenhouse gas emissions," Biomass and Bioenergy 28 (2005) 475-489.
Progress 01/01/04 to 12/31/04
I attended the 2004 annual meeting of the Multistate Research Project S-1007 in Golden, CO, Sept 30-Oct. 1. There, as Chair of Objective 5, Education and Outreach, I made a presentation describing educational initiatives. I summarized research activities from Michigan related to biobased products and discussed development of a joint research proposal with Florida S-1007 representatives. In 2004, I continued to serve as webmaster for the S-1007 group. The website I set up is at the URL (www.egr.msu.edu/bio/srdc). I posted several electronic presentations from the 2004 meeting, and earlier S-1007 meetings on that site.
This proposed research collaboration between Florida and Michigan would integrate the metabolic engineering expertise of Lonnie Ingram's lab (Florida), with the enzyme engineering expertise of Greg Zeikus' lab (Michigan) and expertise in nanostructured biomimetic interfaces of Mark Worden's lab (Michigan). The expected result would be new biological fuel cells to convert biobased fuels into electricity or biocatalytic reactors to convert biobased chemicals into higher-value products. Objective 5: Education and Outreach of the Multiregional Project is intended to help develop the skilled labor force needed for the fledgling biobased products industry. The National Resource Center on Biomass Education would develop and/or organize educational materials and make them broadly available to the public. Emphasis will be placed on development of distance-education materials to provide access to students at schools without a concentration of faculty having expertise in
technologies for a biobased economy.
- Kohli, N., P.R. Dvornic, S.N. Kaganove, R.M. Worden, and I. Lee (2004) "Nanostructured Cross-linkable Micropatterns via Amphiphilic Dendrimer Stamping, Macromolecular Rapid Communications 25, 935 to 941. (Cover article)
- Kohli, N., R.M. Worden, and I. Lee (2005) Intact Transfer of Layered, Bionanocomposite Arrays by Microcontact Printing, Chemical Communications, 2005, 3, 316 to 318.
Progress 01/01/03 to 12/31/03
This year, I served as Chair of Objective 5: Education and Outreach. I also served as webmaster for the Multiregional Project. The website I created is at the URL (http://www.egr.msu.edu/bio/srdc/index.html). I attended the annual meeting of the Multiregional Project Nov. 6-7, 2003 in Washington DC and made an oral presentation descibing plans to develop a National Resource Center on Biomass Education (NRCBE).
Objective 5: Education and Outreach of the Multiregional Project is intended to help develop the skilled labor force needed for the fledgling biobased products industry. The National Resource Center on Biomass Education would develop and/or organize educational materials and make them broadly available to the public. Emphasis will be placed on development of distance-education materials to provide access to students at schools without a concentration of faculty having expertise in technologies for a biobased economy.
- No publications reported this period