Life Sciences Commons^™

Soil Respiration Measurements Reveal High Retention Of Organic Carbon From Corn Residue Derived High-Lignin Fermentation Byproduct Enabling Sustainable Lignocellulosic Biofuel Production, Michelle Sun Wang

Dartmouth College Master’s Theses

While 2G biofuel production can utilize non-edible, lignocellulosic feedstocks such as agricultural residues to produce liquid fuel, harvesting crop residues is unsustainable without careful management of the soil underneath. By harvesting a fraction of the crop residues left in the field after harvest, soil health can diminish and critically, the soil organic carbon (SOC) stored in agricultural fields can decrease. Currently, in the most popular 2G process models published, the issue of soil degradation remains unresolved with residue harvest strategies receiving considerable attention in the literature and other SOC management strategies receiving far less. Specifically, the strategy of returning the …

Lipid Droplet Membrane Proteome Remodeling Parallels Ethanol-Induced Hepatic Steatosis And Its Resolution, Carol A. Casey, Terrence M. Donohue Jr., Jacy L. Kubik, Vikas Kumar, Michael J. Naldrett, Nicholas T. Woods, Cole P. Frisbie, Mark A. Mcniven, Paul G. Thomes

Nebraska Center for Biotechnology: Faculty and Staff Publications

Abstract Lipid droplets (LDs) are composed of neutral lipids enclosed in a phospholipid monolayer, which harbors membrane-associated proteins that regulate LD functions. Despite the crucial role of LDs in lipid metabolism, remodeling of LD protein composition in disease contexts, such as steatosis, remains poorly understood. We hypothesized that chronic ethanol consumption, subsequent abstinence from ethanol, or fasting differentially affects the LD membrane proteome content and that these changes influence how LDs interact with other intracellular organelles. Here, male Wistar rats were pair-fed liquid control or ethanol diets for 6 weeks, and then, randomly chosen animals from both groups were either …

Our Energy Future, Fall/Winter 2007, Issue 15

Sustain Magazine

No abstract provided.

Both Adhe And A Separate Nadph-Dependent Alcohol Dehydrogenase Gene, Adha, Are Necessary For High Ethanol Production In Thermoanaerobacterium Saccharolyticum, Tianyong Zheng, Daniel G. Olson, Sean J. Murphy, Xiongjun Shao, Liang Tian, Lee Lynd

Dartmouth Scholarship

Thermoanaerobacterium saccharolyticum has been engineered to produce ethanol at ∼90% theoretical yield and titer of 70 g/L. Its ethanol-producing ability has drawn attention to its metabolic pathways, which could potentially be transferred to other organisms of interest. Here we report that the iron-containing AdhA is important for ethanol production in the high-ethanol strain of T. saccharolyticum (LL1049). A single-gene deletion of adhA in LL1049 reduced ethanol production by ∼50%, whereas multiple gene deletions of all annotated alcohol dehydrogenases except adhA and adhE did not affect ethanol production. Deletion of adhA in wild-type T. saccharolyticum reduced NADPH-linked ADH activity (acetaldehyde-reducing) by …

Engineering, Nutrient Removal, And Feedstock Conversion Evaluations Of Four Corn Stover Harvest Scenarios, Reed L. Hoskinson, Douglas L. Karlen, Stuart J. Birrell, Corey W. Radtke, W. W. Wilhelm

Douglas L Karlen

Crop residue has been identified as a near-term source of biomass for renewable fuel, heat, power, chemicals and other bio-materials. A prototype one-pass harvest system was used to collect residue samples from a corn (Zea mays L.) field near Ames, IA. Four harvest scenarios (low cut, high-cut top, high-cut bottom, and normal cut) were evaluated and are expressed as collected stover harvest indices (CSHI). High-cut top and high-cut bottom samples were obtained from the same plot in separate operations. Chemical composition, dilute acid pretreatment response, ethanol conversion yield and efficiency, and thermochemical conversion for each scenario were determined. Mean grain …

Storage Of Round And Square Switchgrass Bales: Effect Of Storage Time And Fungal Inoculation On Saccharification Efficiency, Noaa Frederick

Graduate Theses and Dissertations

To produce fermentable sugars from lignocellulosic biomass feedstock, severe pretreatment conditions are needed (either high acid concentration, temperature, or retention times). High severities can produce toxic byproducts which inhibit enzymatic hydrolysis or fermentation. In order to reduce pretreatment severities (and thus increase enzyme and fermentation efficiency), the white-rot fungus Pleurotus ostreastus was seeded into square and round bales of Kanlow switchgrass (Panicum virgastum L.) and left in the field over a period of 9 month. The laccase producing fungus is believed to selectively degrade lignin, a common plant structural polymer, which can function as an enzymatic inhibitor. Samples were taken …

Deletion Of Nfnab In Thermoanaerobacterium Saccharolyticum And Its Effect On Metabolism, Jonathan Lo, Tianyong Zheng, Daniel G. Olson, Natalie Ruppertsberger, Shital Tripathi, Adam Guss, Lee Lynd

Dartmouth Scholarship

NfnAB catalyzes the reversible transfer of electrons from reduced ferredoxin and NADH to 2 NADP⁺. The NfnAB complex has been hypothesized to be the main enzyme for ferredoxin oxidization in strains of Thermoanaerobacterium saccharolyticum engineered for increased ethanol production. NfnAB complex activity was detectable in crude cell extracts of T. saccharolyticum. Activity was also detected using activity staining of native PAGE gels. The nfnAB gene was deleted in different strains of T. saccharolyticum to determine its effect on end product formation. In wild-type T. saccharolyticum, deletion of nfnAB resulted in a 46% increase in H₂ formation but …

Elimination Of Hydrogenase Active Site Assembly Blocks H2 Production And Increases Ethanol Yield In Clostridium Thermocellum, Ranjita Biswas, Tianyong Zheng, Daniel G. Olson, Lee R. Lynd, Adam M. Guss

Dartmouth Scholarship

Background: The native ability of Clostridium thermocellum to rapidly consume cellulose and produce ethanol makes it a leading candidate for a consolidated bioprocessing (CBP) biofuel production strategy. C. thermocellum also synthesizes lactate, formate, acetate, H₂ , and amino acids that compete with ethanol production for carbon and electrons. Elimination of H₂ production could redirect carbon flux towards ethanol production by making more electrons available for acetyl coenzyme A reduction to ethanol. Results: H₂ production in C. thermocellum is encoded by four hydrogenases. Rather than delete each individually, we targeted hydrogenase maturase gene hydG, involved in converting the …

The Bifunctional Alcohol And Aldehyde Dehydrogenase Gene, Adhe, Is Necessary For Ethanol Production In Clostridium Thermocellum And Thermoanaerobacterium Saccharolyticum, Jonathan Lo, Tianyong Zheng, Shuen Hon, Daniel G. Olson, Lee Lynd

Dartmouth Scholarship

Thermoanaerobacterium saccharolyticum and Clostridium thermocellum are anaerobic thermophilic bacteria being investigated for their ability to produce biofuels from plant biomass. The bifunctional alcohol and aldehyde dehydrogenase gene, adhE, is present in these bacteria and has been known to be important for ethanol formation in other anaerobic alcohol producers. This study explores the inactivation of the adhE gene in C. thermocellum and T. saccharolyticum. Deletion of adhE reduced ethanol production by >95% in both T. saccharolyticum and C. thermocellum, confirming that adhE is necessary for ethanol formation in both organisms. In both adhE deletion strains, fermentation products shifted from ethanol …

Developing An Unstructured Model To Investigate The Effect Of Ethanol On Product Yields For Glucose And Xylose Cofermentation In Saccharomyces Cerevisiae 424a (Lnh-St), Shane D. Clingenpeel

Open Access Theses

Production of bio-ethanol from lignocellulose requires the efficient fermentation of glucose and xylose, even in the presence of inhibitors. The desired product, ethanol itself, will inhibit the fermentation. A further understanding of how ethanol affects the organism is critical to overcoming its inhibition.

This thesis evaluated the effect of ethanol on the cofermentation of glucose and xylose in two different cases. The first case had an unstructured model created for Saccharomyces cerevisiae 424A (LNH-ST), a genetically modified strain of yeast capable of cofermenting glucose and xylose. The differential equations were based around sugar consumption, and the product yields were investigated …

Coordinated Dynamic Gene Expression Changes In The Central Nucleus Of The Amygdala During Alcohol Withdrawal., Kate Freeman, Mary M. Staehle, Rajanikanth Vadigepalli, Gregory E Gonye, Babatunde A Ogunnaike, Jan B Hoek, James S Schwaber

Faculty Scholarship for the College of Science & Mathematics

BACKGROUND: Chronic alcohol use causes widespread changes in the cellular biology of the amygdala's central nucleus (CeA), a GABAergic center that integrates autonomic physiology with the emotional aspects of motivation and learning. While alcohol-induced neurochemical changes play a role in dependence and drinking behavior, little is known about the CeA's dynamic changes during withdrawal, a period of emotional and physiologic disturbance.

METHODS: We used a qRT-PCR platform to measure 139 transcripts in 92 rat CeA samples from control (N = 33), chronically alcohol exposed (N = 26), and withdrawn rats (t = 4, 8, 18, 32, and 48 hours; N …

High Ethanol Titers From Cellulose By Using Metabolically Engineered Thermophilic, Anaerobic Microbes, D. Aaron Argyros, Shital A. Tripathi, Trisha F. Barrett, Stephen R. Rogers, Lawrence F. Feinberg, Daniel G. Olson, Justin M. Foden, Bethany B. Miller, Lee R. Lynd, David A. Hogsett, Nicky C. Caiazza

Dartmouth Scholarship

This work describes novel genetic tools for use in Clostridium thermocellum that allow creation of unmarked mutations while using a replicating plasmid. The strategy employed counter-selections developed from the native C. thermocellum hpt gene and the Thermoanaerobacterium saccharolyticum tdk gene and was used to delete the genes for both lactate dehydrogenase (Ldh) and phosphotransacetylase (Pta). The Δldh Δpta mutant was evolved for 2,000 h, resulting in a stable strain with 40:1 ethanol selectivity and a 4.2-fold increase in ethanol yield over the wild-type strain. Ethanol production from cellulose was investigated with an engineered coculture of organic acid-deficient engineered strains of …

Mutant Alcohol Dehydrogenase Leads To Improved Ethanol Tolerance In Clostridium Thermocellum, Steven D. Brown, Adam M. Guss, Tatiana V. Karpinets, Jerry M. Parks

Dartmouth Scholarship

Clostridium thermocellum is a thermophilic, obligately anaerobic, Gram-positive bacterium that is a candidate microorganism for converting cellulosic biomass into ethanol through consolidated bioprocessing. Ethanol intolerance is an important metric in terms of process economics, and tolerance has often been described as a complex and likely multigenic trait for which complex gene interactions come into play. Here, we resequence the genome of an ethanol-tolerant mutant, show that the tolerant phenotype is primarily due to a mutated bifunctional acetaldehyde-CoA/alcohol dehydrogenase gene (adhE), hypothesize based on structural analysis that cofactor specificity may be affected, and confirm this hypothesis using enzyme assays. …

Identification Of The [Fefe]-Hydrogenase Responsible For Hydrogen Generation In Thermoanaerobacterium Saccharolyticum And Demonstration Of Increased Ethanol Yield Via Hydrogenase Knockout, A. Joe Shaw, David A. Hogsett, Lee R. Lynd

Dartmouth Scholarship

Three putative hydrogenase enzyme systems in Thermoanaerobacterium saccharolyticum were investigated at the genetic, mRNA, enzymatic, and phenotypic levels. A four-gene operon containing two [FeFe]-hydrogenase genes, provisionally termed hfs (hydrogenase-Fe-S), was found to be the main enzymatic catalyst of hydrogen production. hfsB, perhaps the most interesting gene of the operon, contains an [FeFe]-hydrogenase and a PAS sensory domain and has several conserved homologues among clostridial saccharolytic, cellulolytic, and pathogenic bacteria. A second hydrogenase gene cluster, hyd, exhibited methyl viologen-linked hydrogenase enzymatic activity, but hyd gene knockouts did not influence the hydrogen yield of …

Temperature Influence And Heat Management Requirements Of Microalgae Cultivation In Photobioreactors, Thomas Hagen Mehlitz

Master's Theses

Microalgae are considered one of the most promising feedstocks for biofuel production for the future. The most efficient way to produce vast amounts of algal biomass is the use of closed tubular photobioreactors (PBR). The heat requirement for a given system is a major concern since the best algae growth rates are obtained between 25-30 °C, depending on the specific strain. A procedure to determine temperature influence on algal growth rates was developed for a lab-scale PBR system using the species Chlorella. A maximum growth rate of 1.44 doublings per day at 29 °C (optimal temperature) was determined. In addition, …

Quantifying Total And Sustainable Agricultural Biomass Resources In South Dakota—A Preliminary Assessment, Kurt A. Rosentrater, Dennis Todey, Russell Persyn

Kurt A. Rosentrater

Conversion of biomass is considered the next major advance in biorenewable fuels, energy, and products. Wholesale conversion to biomass utilization could result in removal of current crop residues from agricultural fields (on prime agricultural lands) or even implementation of different crops and cropping strategies (i.e., switchgrass on marginal lands). To date, the driver for biomass processing has been economics and limitations on the conversion of the lignocellulose. Over the last forty years significant investments and resultant changes in management practices in the agricultural sector have focused on soil and water conservation. One of the major efforts has focused on conservation-till …

Metabolic Engineering Of A Thermophilic Bacterium To Produce Ethanol At High Yield, A. Joe Shaw, Kara K. Podkaminer, Sunil G. Desai, John S. Bardsley, Stephen R. Rogers, Philip G. Thorne, David A. Hogsett, Lee R. Lynd

Dartmouth Scholarship

We report engineering Thermoanaerobacterium saccharolyticum, a thermophilic anaerobic bacterium that ferments xylan and biomass-derived sugars, to produce ethanol at high yield. Knockout of genes involved in organic acid formation (acetate kinase, phosphate acetyltransferase, and L-lactate dehydrogenase) resulted in a strain able to produce ethanol as the only detectable organic product and substantial changes in electron flow relative to the wild type. Ethanol formation in the engineered strain (ALK2) utilizes pyruvate:ferredoxin oxidoreductase with electrons transferred from ferredoxin to NAD(P), a pathway different from that in previously described microbes with a homoethanol fermentation. The homoethanologenic phenotype was stable for >150 generations …

Physical And Chemical Properties Of Corn Distillers Wet Grains, Kurt A. Rosentrater, R. Michael Lehman

Kurt A. Rosentrater

The production of corn-based ethanol in the United States is dramatically increasing, and consequently, so is the quantity of generated coproducts, all of which are currently fed to livestock. Distillers Wet Grain (DWG) is one of these coproduct feeds. Many studies have investigated the nutritional properties of DWG and the response of livestock (primarily beef and dairy) to inclusion of these ingredients in their diets. To date, physical properties have been largely ignored, but they are needed for the design of storage facilities and equipment. The objective of this research was to quantify physical and chemical property values for typical …

Biomass And Biofuels In Maine: Estimating Supplies For Expanding The Forest Products Industry, Jonathan Rubin, Kate Dickerson, Jacob Kavkewitz

Energy & the Environment

This paper estimates the renewable energy potential of Maine’s forest resources, and how much energy these resources could potentially provide the state. Using the most recent state-specific data available, and a methodology similar to the Billion Tons Report, we find that ethanol production from Maine’s forest residues could potentially provide 18% of Maine’s transportation (gasoline) fuels with a fermentation wood to ethanol process. Making Fischer-Tropsch diesel (F-T diesel) using forest residues can replace 39% of Maine’s petro-diesel consumption. Actual levels of biofuels that can be produced will depend on conversion factors and forestry residue removals that are subject to uncertainty.

Potential Bleaching Techniques For Use In Distillers Grains, Jessica A. Saunders, Kurt A. Rosentrater, Padmanaban G. Krishnan

Kurt A. Rosentrater

The ethanol industry is booming. And extensive research is currently being pursued to develop alternative uses for distillers dried grains (DDG) and distillers dried grains with solubles (DDGS), coproducts of the ethanol production process. Currently, DDG and DDGS are used exclusively as livestock feed. previous research has shown promising avenues for the use of DDG and DDGS in human foods. The low starch, high protein and high fiber grains seem ideally suited for medical conditions such as diabetes and Celiac's disease. Processing methods need to be investigated to create functional flours that can be used effectively. Refining aspects such as …

Quantifying Total And Sustainable Agricultural Biomass Resources In South Dakota, Kurt A. Rosentrater, Russell Persyn, Dennis Todey

Kurt A. Rosentrater

Conversion of biomass is considered the next major advance in biorenewable fuels, energy, and products. Wholesale conversion to biomass utilization could result in removal of current crop residues from agricultural fields or even implementation of different crops and cropping strategies (i.e., switchgrass). To date, the driver for biomass processing has been economics and limitations on the conversion of the lignocellulose. Over the last forty years significant investments and resultant changes in management practices in the agricultural sector have focused on soil and water conservation. One of the major efforts has focused on conservation-till or no-till, with the goal of retaining …

Physical Properties Of Low Oil Distillers Dried Grains With Solubles (Ddgs), Jessica A. Herr Saunders, Kurt A. Rosentrater

Kurt A. Rosentrater

Corn-based ethanol production is exponentially growing in the U.S. As the use of ethanol as a fuel source increases, so does the need to find valuable uses for coproducts of the production process, such as distillers dried grains with solubles (DDGS). DDGS is a good source of nutrients, energy, and protein. Currently DDGS is sold by ethanol processors as livestock feed, thereby increasing profit for the production plants. As the industry continues to grow, there will be a growing need to find additional uses for DDGS. Physical and chemical properties of coproduct streams are becoming increasingly investigated, as these characteristics …

Characterization Of Chemical And Physical Properties Of Distillers Dried Grain With Solubles (Ddgs) For Value Added Uses, Rumela Bhadra, K. Muthukumarappan, Kurt A. Rosentrater

Kurt A. Rosentrater

One of the fastest growing industries in the United States is the fuel ethanol industry. Since 2000 there has been an increase of more than 300%. There was production of 4.9 billion gallons of ethanol in 2006. The major coproducts from this industry include Distillers Dried Grains with Solubles (DDGS) and carbon dioxide. DDGS is used as a livestock feed since it contains high quantities of protein, fiber, amino acids, and other nutrients. The goal of this study was to quantify various chemical and physical properties of DDGS, Distillers Wet Grains (DWG), and Distillers Dried Grain (DDG) from several plants …

Economic Analysis Of Cellulase Production Methods For Bio-Ethanol, Jun Zhuang, Mary A. Marchant, Sue E. Nokes, Herbert J. Strobel

Biosystems and Agricultural Engineering Faculty Publications

The cost of cellulase enzymes has limited the feasibility of producing ethanol from fibrous biomass. Traditional submerged fermentation (SmF) was compared to an alternative method of producing cellulase, solid state cultivation (SSC). Results from an economic analysis indicated that the unit costs for cellulase enzyme production were $15.67 (The prices are all 2004 prices in this article, except otherwise stated. We deflated newer prices to 2004 prices using a deflation factor 0.9 per year and inflated older prices to 2004 prices using an inflation factor 1.1.) per kilogram ($/kg) and $40.36/kg, for the SSC and SmF methods, respectively, while the …

Physical Properties Of Distillers Dried Grains With Solubles (Ddgs), Kurt A. Rosentrater

Kurt A. Rosentrater

The production of corn-based ethanol in the U.S. is dramatically increasing, and consequently so is the amount of coproduct materials generated from this processing sector. These streams are primarily utilized as livestock feed, which is a route that provides ethanol processors with a substantial revenue source and significantly increases the profitability of the production process. With the construction and operation of many new plants in recent years, these residuals do, however, have much potential for value-added processing and utilization in other sectors as well. Extensive research has been conducted into determining the nutritional properties of distillers dried grains with solubles …

Anaerobic Digestion Potential For Ethanol Processing Residues, Kurt A. Rosentrater, Heath R. Hall, Conly L. Hansen

Kurt A. Rosentrater

The production of corn-based ethanol in the U.S. is dramatically increasing, and consequently so is the quantity of byproduct materials generated from this processing sector. These coproduct streams are currently solely utilized as livestock feed, which is a route that provides ethanol processors with a substantial revenue source and significantly increases the profitability of the production process. With the construction and operation of many new plants in recent years, these residuals do, however, have much potential for value-added processing and utilization in other sectors as well. This option holds the promise of economic benefit for corn processors, especially if the …

Fractionation Techniques To Concentrate Nutrient Streams In Distillers Grains, Kurt A. Rosentrater, Deepa Subramanian, Padmanaban G. Krishnan

Kurt A. Rosentrater

Corn, the most widely produced feed grain in the United States, accounts for more than 90% of the total value and production of feed grains. It is also used for food, industrial materials, and fuel ethanol production. Distillers grains, the major coproduct from ethanol manufacturing, are used as livestock feed. There are, however, other potential options, including value-added food, industrial, chemical, and energy applications. Fractionating distillers grains into concentrated streams of protein, fiber, and fat may be key to facilitating these types of utilization. Previous studies on other products have shown that fractionation is a promising mechanism for extracting valuable …

Methodology To Determine Soluble Content In Dry Grind Ethanol Coproduct Streams, Vykundeshwari Ganesan, Kurt A. Rosentrater, Kasiviswanathan Muthukumarappan

Kurt A. Rosentrater

Distillers grains and syrup are coproducts from fuel ethanol dry grind processing. Ethanol manufacturing is dramatically increasing in the United States, primarily in Midwestern states, and thus the availability of these feed products is also growing. Confusion currently exists in industrial nomenclature regarding "solubles" in these streams because no standards are in place. In our study, dissolved materials were considered soluble matter. We developed a methodology to determine the dry basis soluble content in condensed distillers solubles (CDS) and distillers dried grains with solubles (DDGS). A mass balance analytical approach was initially used, but results were not in good agreement …

Some Physical Properties Of Distillers Dried Grains With Solubles (Ddgs), Kurt A. Rosentrater

Kurt A. Rosentrater

With the rapid growth in the fuel ethanol industry in recent years, considerable research is being devoted to determining distillers dried grains with solubles (DDGS) nutritional properties and to optimizing their inclusion in livestock diets; physical properties of these materials, however, have been largely ignored. Using standard laboratory methods, several physical properties for typical DDGS streams were determined, including moisture content, water activity, thermal properties (conductivity, resistivity, and diffusivity), bulk density, angle of repose, and color. The DDGS samples in this study were golden-brown in color and exhibited physical properties similar to other dry feed ingredients, such as hominy feed, …