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At least 1,333 records · Page 74
All sourcesNASA NTRSDOE OSTI
DOE OSTI2019

Systems-Level Analysis of Oxygen Exposure in Zymomonas mobilis : Implications for Isoprenoid Production

ABSTRACT Zymomonas mobilis is an aerotolerant anaerobe and prolific ethanologen with attractive characteristics for industrial bioproduct generation. However, there is currently insufficient knowledge of the impact that environmental factors have on flux through industrially relevant biosynthetic pathways. Here, we examined the effect of oxygen exposure on metabolism and gene expression in Z. mobilis by combining targeted metabolomics, mRNA sequencing, and shotgun proteomics. We found that exposure to oxygen profoundly influenced metabolism, inducing both transient metabolic bottlenecks and long-term metabolic remodeling. In particular, oxygen induced a severe but temporary metabolic bottleneck in the methyl erythritol 4-phosphate pathway for isoprenoid biosynthesis caused by oxidative damage to the iron-sulfur cofactors of the final two enzymes in the pathway. This bottleneck was resolved with minimal changes in expression of isoprenoid biosynthetic enzymes. Instead, it was associated with pronounced upregulation of enzymes related to iron-sulfur cluster maintenance and biogenesis (i.e., flavodoxin reductase and the suf operon). We also detected major changes in glucose utilization in the presence of oxygen. Specifically, we observed increased gluconate production following exposure to oxygen, accounting for 18% of glucose uptake. Our results suggest that under aerobic conditions, electrons derived from the oxidation of glucose to gluconate are diverted to the electron transport chain, where they can minimize oxidative damage by reducing reactive oxygen species such as H 2 O 2 . This model is supported by the simultaneous upregulation of three membrane-bound dehydrogenases, cytochrome c peroxidase, and a cytochrome bd oxidase following exposure to oxygen. IMPORTANCE Microbially generated biofuels and bioproducts have the potential to provide a more environmentally sustainable alternative to fossil-fuel-derived products. In particular, isoprenoids, a diverse class of natural products, are chemically suitable for use as high-grade transport fuels and other commodity molecules. However, metabolic engineering for increased production of isoprenoids and other bioproducts is limited by an incomplete understanding of factors that control flux through biosynthetic pathways. Here, we examined the native regulation of the isoprenoid biosynthetic pathway in the biofuel producer Zymomonas mobilis . We leveraged oxygen exposure as a means to perturb carbon flux, allowing us to observe the formation and resolution of a metabolic bottleneck in the pathway. Our multi-omics analysis of this perturbation enabled us to identify key auxiliary enzymes whose expression correlates with increased production of isoprenoid precursors, which we propose as potential targets for future metabolic engineering.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Urban Energy and Climate: Prospects for a Sustainable Transition

With the continuous migration of people towards metropolitan areas in search of employment, the demands for core services and energy, coupled with an increasing awareness of the impact of climate change, have placed the management and planning of global urban energy under a lot of pressure. Trends toward urban energy service transformations that offer greater affordability, reliability, efficiency and adaptability provide hope for a global sustainable future. At the same time, there are also limits to these transitions, as well as risks involved. For example, on one end of the spectrum, our urban energy future includes land use sprawl, high fossil fuel use, pollution, and unhealthy urban conditions. On the other side of this transition spectrum is more energy choices, and healthier, more livable cities, along with less energy use and fewer greenhouse gas emissions. What the future might hold for transforming the world's cities depends upon an understanding of the risks of current trajectories and the opportunities for and limitations to developing sustainable urban energy systems. This edited volume brings together leading experts on the prospects and challenges of urban energy innovation and on related-economic, social and environmental sustainability transitions. The focus of the volume is on multidisciplinary reviews, research informing technologies and policies for sustainability, and analytical insights addressing rapid urbanization and changes across a diverse typology of global cities. The volume will include an overview of the current state of urban energy systems. It will also document and evaluate urban energy prospects for a sustainable, resilient future.

drivers of change↗
DOE OSTI2013

A MULTI-MODEL ANALYSIS OF POST-2020 MITIGATION EFFORTS OF FIVE MAJOR ECONOMIES

This paper looks into the regional mitigation strategies of five major economies (China, EU, India, Japan, and USA) in the context of the 2°C target, using a multi-model comparison. In order to stay in line with the 2°C target, a tripling or quadrupling of mitigation ambitions is required in all regions by 2050, employing vigorous decarbonization of the energy supply system and achieving negative emissions during the second half of the century. In all regions looked at, decarbonization of energy supply (and in particular power generation) is more important than reducing energy demand. Some differences in abatement strategies across the regions are projected: In India and the USA the emphasis is on prolonging fossil fuel use by coupling conventional technologies with carbon storage, whereas the other main strategy depicts a shift to carbon-neutral technologies with mostly renewables (China, EU) or nuclear power (Japan). Regions with access to large amounts of biomass, such as the USA, China, and the EU, can make a trade-off between energy related emissions and land related emissions, as the use of bioenergy can lead to a net increase in land use emissions. After supply-side changes, the most important abatement strategy focuses on end-use efficiency improvements, leading to considerable emission reductions in both the industry and transport sectors across all regions. Abatement strategies for non-CO2 emissions and land use emissions are found to have a smaller potential. Inherent model, as well as collective, biases have been observed affecting the regional response strategy or the available reduction potential in specific (end-use) sectors.

abatement↗
DOE OSTI2015

Data Center Energy Efficiency Standards in India: Preliminary Findings from Global Practices

Global data center energy consumption is growing rapidly. In India, information technology industry growth, fossil-fuel generation, and rising energy prices add significant operational costs and carbon emissions from energy-intensive data centers. Adoption of energy-efficient practices can improve the global competitiveness and sustainability of data centers in India. Previous studies have concluded that advancement of energy efficiency standards through policy and regulatory mechanisms is the fastest path to accelerate the adoption of energy-efficient practices in the Indian data centers. In this study, we reviewed data center energy efficiency practices in the United States, Europe, and Asia. Using evaluation metrics, we identified an initial set of energy efficiency standards applicable to the Indian context using the existing policy mechanisms. These preliminary findings support next steps to recommend energy efficiency standards and inform policy makers on strategies to adopt energy-efficient technologies and practices in Indian data centers.

Raje, Sanyukta↗
DOE OSTI2017

A Data Driven Pre-cooling Framework for Energy Cost Optimization in Commercial Buildings

Commercial buildings consume significant amount of energy. Facility managers are increasingly grappling with the problem of reducing their buildings’ peak power, overall energy consumption and energy bills. In this paper, we first develop an optimization framework – based on a gray box model for zone thermal dynamics – to determine a pre-cooling strategy that simultaneously shifts the peak power to low energy tariff regimes, and reduces both the peak power and overall energy consumption by exploiting the flexibility in a building’s thermal comfort range. We then evaluate the efficacy of the pre-cooling optimization framework by applying it to building management system data, spanning several days, obtained from a large commercial building located in a tropical region of the world. The results from simulations show that optimal pre-cooling reduces peak power by over 50%, energy consumption by up to 30% and energy bills by up to 37%. Next, to enable ease of use of our framework, we also propose a shortest path based heuristic algorithmfor solving the optimization problemand show that it has comparable erformance with the optimal solution. Finally, we describe an application of the proposed optimization framework for developing countries to reduce the dependency on expensive fossil fuels, which are often used as a source for energy backup.We conclude by highlighting our real world deployment of the optimal pre-cooling framework via a software service on the cloud platform of a major provider. Our pre-cooling methodology, based on the gray box optimization framework, incurs no capital expense and relies on data readily available from a building management system, thus enabling facility managers to take informed decisions for improving the energy and cost footprints of their buildings

Vishwanath, Arun↗
DOE OSTI2022

Control of Low-Inertia Power Systems

Electric power systems are undergoing an unprecedented transition from fossil fuel–based power plants to low-inertia systems that rely predominantly on power electronics and renewable energy resources. This article reviews the resulting control challenges and modeling fallacies, at both the device and system level, and focuses on novel aspects or classical concepts that need to be revised in light of the transition to low-inertia systems. To this end, we survey the literature on modeling of low-inertia systems, review research on the control of grid-connected power converters, and discuss the frequency dynamics of low-inertia systems. Moreover, we discuss system-level services from a control perspective. Overall, we conclude that the system-theoretic mindset is essential to bridge different research communities and understand the complex interactions of power electronics, electric machines, and their controls in large-scale low-inertia power systems.

24 POWER TRANSMISSION AND DISTRIBUTION↗
DOE OSTI2022

Hydrogen and Battery Storage Technologies for Low-Cost Energy Decarbonization in Distribution Networks

Deep energy decarbonization cannot be achieved without high penetration of renewables. At higher renewable energy penetrations, the variability and intermittent nature of solar photovoltaic (PV) electricity can cause ramping issues with existing fossil fuel generation, requiring longer term energy storage to increase the reliability of grid operation. Here, a proton exchange membrane electrolyzer can produce H 2 and serves as a utility controllable load. The produced H 2 can then be stored and converted back into electricity, or mixed with natural gas, or used as transportation fuel, or chemical feedstock. This paper considers the perspective of the distribution system operator that operates the distributed energy resources on a standard IEEE 33-node distribution network considering the technical and physical constraints with the goal of minimizing total investment and operation cost. Different case studies, at very high PV penetrations are considered to show the challenges and path to net-zero emission energy production using H 2 energy. Sensitivity of utility PV costs and electrolyzer capital costs on producing H 2 at $\$1$ /kg are presented showing that the distribution network could produce 100% renewable electricity and H 2 could be produced at a cost of $\$1$ /kg by 2050 with conservative cost estimates and by 2030 with accelerated cost declines.

25 ENERGY STORAGE↗
DOE OSTI2015

The Selective Electrochemical Conversion of Preactivated CO 2 to Methane

This work reports the selective electrochemical conversion of CO 2 to methane, the reverse reaction of fossil fuel combustion. This reaction is facilitated by preactivation of the CO 2 molecule with an N-heterocyclic carbene (NHC) to form a zwitterionic species in the first step. In the presence of Ni(cyclam) 2+ and CF 3 CH 2 OH, this species is shown to undergo further electrochemical reduction of the bound-CO 2 fragment at glassy carbon cathodes in dichloromethane electrolyte solution. Labeling studies confirm the origin of the carbon and protons in the methane product are the preactivated CO 2 and trifluoroethanol respectively.

30 DIRECT ENERGY CONVERSION↗
DOE OSTI2021

Chalkboard 2 - How to Make Clean Hydrogen

Clean hydrogen is a carbon-free energy carrier that can be produced from water and sustainable energy sources such as wind, solar, and nuclear. Therefore, clean hydrogen is one of the best ways to not only decarbonize the energy supply system, but also address the zero-emission challenges specific to large-carbon emitting industries that are difficult to separate from fossil fuels. To help achieve the Biden Administration's goal of a 100% clean energy economy and net-zero emissions by 2050, several tens of millions of metric tons of clean, low-cost hydrogen will be needed annually. The HydroGEN Advanced Water Splitting Materials (AWSM) Consortium was established in 2016 as part of the Energy Materials Network (EMN) under the U.S. Department of Energy, Office of Energy Efficiency and Renewable Energy, Hydrogen and Fuel Cell Technologies Office to enhance the performance, improve the durability, and reduce the cost of clean hydrogen production technologies, and it is helping to advance the H2@Scale vision.

08 HYDROGEN↗
DOE OSTI2022

Co-simulation Framework for Community-scale Building-grid Integration [SWR-21-75]

Distributed energy resources (DERs), including rooftop solar, energy storage, and flexible loads, are gaining popularity as costs decline and as building owners and utilities realize their benefits. DERs can improve distribution system efficiency, help prevent the need for expensive grid upgrades, and increase the resilience of local communities. However, they can also cause difficulties in grid operations and can require controls to achieve their benefits. To address this challenge, NREL researchers have developed a community-scale solution that assesses the impacts of DERs and their control strategies on a distribution system. The framework has been shown to reduce solar photovoltaic (PV) curtailment to 0%, mitigate the adverse impact of solar variability on the distribution voltage, and provide up to 5-day critical load support during emergency events. Utilizing 5 different modules representing the feeder, buildings, home energy management systems, an aggregator, and a utility controller, NREL expects this simulation technology to play a critical role in the continued integration of DERs. According to the Energy Information Administration (EIA), solar curtailments accounted for 94% of the total energy curtailed in the California Independent System Operator (CAISO) in 2020. By enabling Independent System Operators (ISOs) and utility operators to bring solar curtailments to 0%, the electrical grid can become less dependent on fossil-fueled power generation sources. NREL's co-simulation framework contains five major components: Distribution Feeder Model: describes the distribution feeder topology using OpenDSS, including the locations of all DERs. Residential Building Model: simulates a large number of buildings at a high resolution using OCHRETM. The model is equipped to control equipment based on signals from an external module. The model includes major household appliances such as HVAC and a water heater, non-dispatchable load models, a distributed PV system, and a home battery system. Home Energy Management System: optimizes the controls for the devices in a home using foreseeTM. The control can adjust based on the user preferences including cost, comfort, and convenience. In hierarchical control scenarios, where the houses follow signals from an aggregator, the home energy management system provides a flexibility band with a range of power and follows the dispatch signals received from aggregator. Community-Level Aggregator: solves for optimal energy dispatch based on the flexibility bands received from each home and the grid service signal received from the utility controller. Utility-Level Controller: provides grid signals for voltage control using Distributed Energy Resources (DERs), such as solar systems, in the community.

Balamurugan, Sivasathya Pradha↗
DOE OSTI2020

The Effects of Synthesis Conditions on the Carbon Capture Capacity of Polyethylenimine Impregnated Protonated Titanate Nanotubes

Capture and storage of carbon dioxide emitted from power plants, which burn fossil fuels, is one of the best ways to mitigate climate change. Polyethylenimine (PEI) impregnated protonated titanate nanotubes (PEI-PTNTs) are efficient amine-modified solid adsorbents for post combustion carbon capture. This study is to investigate the effects of synthesis conditions of titanate nanotubes on the carbon capture capacity of PEI-PTNTs. PTNTs are synthesized using hydrothermal treatment of TiO 2 anatase powder at different temperatures and in different days, followed by washing with dilute HCl solution and deionized water. PEI-PTNTs are prepared by loading PEI (M.W. = 10,000) onto PTNTs at a weight ratio of 1:1 using a wet impregnation process. PTNTs and PEI-PTNTs are characterized using transmission electron microscopy, porosity and surface area analysis, powder X-ray diffraction, and Fourier transform infrared spectroscopy. CO 2 adsorption is conducted at 75 °C using simulated flue gas, and CO 2 desorption is performed at 110 °C by purging the adsorbent with dry nitrogen gas. CO 2 adsorption experiments demonstrate that the PEI-PTNTs synthesized at 130 °C within 3 or 5 days, have the highest CO 2 capture capacity. These optimized PEI-PTNTs have a significant potential to capture CO 2 from power plants.

Materials Science↗
DOE OSTI2018

CO 2 and Carbon Emissions from Cities: Linkages to Air Quality, Socioeconomic Activity, and Stakeholders in the Salt Lake City Urban Area

Urban areas are responsible for a substantial proportion of anthropogenic carbon emissions around the world. As global populations increasingly reside in cities, the role of urban emissions in determining the future trajectory of carbon emissions is magnified. Consequently, a number of research efforts have been started in the United States and beyond, focusing on observing atmospheric carbon dioxide (CO 2 ) and relating its variations to carbon emissions in cities. Because carbon emissions are intimately tied to socioeconomic activity through the combustion of fossil fuels, and many cities are actively adopting emission reduction plans, such urban carbon research efforts give rise to opportunities for stakeholder engagement and guidance on other environmental issues, such as air quality. This paper describes a research effort centered in the Salt Lake City, Utah, metropolitan region, which is the locus for one of the longest-running urban CO 2 networks in the world. The Salt Lake City area provides a rich environment for studying anthropogenic emissions and for understanding the relationship between emissions and socioeconomic activity when the CO 2 observations are enhanced with a) air quality observations, b) novel mobile observations from platforms on light-rail public transit trains and a news helicopter, c) dense meteorological observations, and d) modeling efforts that include atmospheric simulations and high-resolution emission inventories. Carbon dioxide and other atmospheric observations are presented herein, along with associated modeling work. Examples in which the work benefited from and contributed to the interests of multiple stakeholders (e.g., policymakers, air quality managers, municipal government, urban planners, industry, and the general public) are discussed.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2012

Toward a Minimal Representation of Aerosols in Climate Models: Comparative Decomposition of Aerosol Direct, Semidirect, and Indirect Radiative Forcing

The authors have decomposed the anthropogenic aerosol radiative forcing into direct contributions from each aerosol species to the planetary energy balance through absorption and scattering of solar radiation, indirect effects of anthropogenic aerosol on solar and infrared radiation through droplet and crystal nucleation on aerosol, and semidirect effects through the influence of solar absorption on the distribution of clouds. A three-mode representation of the aerosol in version 5.1 of the Community Atmosphere Model (CAM5.1) yields global annual mean radiative forcing estimates for each of these forcing mechanisms that are within 0.1 W m–2 of estimates using a more complex seven-mode representation that distinguishes between fresh and aged black carbon and primary organic matter. Simulating fresh black carbon particles separately from internally mixed accumulation mode particles is found to be important only near fossil fuel sources. In addition to the usual large indirect effect on solar radiation, this study finds an unexpectedly large positive longwave indirect effect (because of enhanced cirrus produced by homogenous nucleation of ice crystals on anthropogenic sulfate), small shortwave and longwave semidirect effects, and a small direct effect (because of cancelation and interactions of direct effects of black carbon and sulfate). Differences between the threemode and seven-mode versions are significantly larger (up to 0.2 W m–2) when the hygroscopicity of primary organic matter is decreased from 0.1 to 0 and transfer of the primary carbonaceous aerosol to the accumulation mode in the seven-mode version requires more hygroscopic material coating the primary particles. Radiative forcing by cloudborne anthropogenic black carbon is only 20.07 W m–2.

Aerosol Direct↗
DOE OSTI2022

Macro Analysis to Estimate Electric Vehicles Fast-Charging Infrastructure Requirements in Small Urban Areas

Electric vehicles (EVs) are known to reduce emissions and fossil fuel dependency. However, the limited range, long charging time, and inadequate charging infrastructure have hampered the adoption of EVs. The current EV charging infrastructure planning studies and tools require detailed information, extensive resources, and skills that can be a significant barrier to urban areas for finding the required charging infrastructure to support a targeted EV market share. This study generates regression models to estimate the number of direct current fast charging stations and the chargers to support the EV charging demand for urban areas. These models provide macro-level estimates of the required infrastructure investment in urban areas, which can be easily implemented by policy-makers and city planners. This study incorporates data obtained from applying a disaggregate optimization-based charger placement model, developed recently by the same authors, for multiple case studies to generate the required data to calibrate the macro-level models, in the state of Michigan. This simulated data set includes the number of charging stations and chargers for each market share, technology advancement scenario, and the transportation network topology. The results show that the number of charging stations reduces with battery size and charging power and increases with EV market share and the road network lane length. The number of chargers reduces with charging power, whereas it increases with battery size, EV market share, and vehicle miles traveled in the system. The model developed here can be applied to any state having urban characteristics and weather conditions similar to Michigan.

Engineering↗
DOE OSTI2019

IJER editorial: The future of the internal combustion engine

Internal combustion (IC) engines operating on fossil fuel oil provide about 25% of the world’s power (about 3000 out of 13,000 million tons oil equivalent per year—see Figure 1), and in doing so, they produce about 10% of the world’s greenhouse gas (GHG) emissions (Figure 2). Reducing fuel consumption and emissions has been the goal of engine researchers and manufacturers for years, as can be seen in the two decades of ground-breaking peer-reviewed articles published in this International Journal of Engine Research (IJER). Indeed, major advances have been made, making today’s IC engine a technological marvel. However, recently, the reputation of IC engines has been dealt a severe blow by emission scandals that threaten the ability of this technology to make significant and further contributions to the reduction of transportation sector emissions. In response, there have been proposals to replace vehicle IC engines with electric-drives with the intended goals of further reducing fuel consumption and emissions, and to decrease vehicle GHG emissions.

33 ADVANCED PROPULSION SYSTEMS↗
DOE OSTI2018

Gene stacking of multiple traits for high yield of fermentable sugars in plant biomass

Second-generation biofuels produced from biomass can help to decrease dependency on fossil fuels, bringing about many economic and environmental benefits. To make biomass more suitable for biorefinery use, we need a better understanding of plant cell wall biosynthesis. Increasing the ratio of C6 to C5 sugars in the cell wall and decreasing the lignin content are two important targets in engineering of plants that are more suitable for downstream processing for second-generation biofuel production. Here, we have studied the basic mechanisms of cell wall biosynthesis and identified genes involved in biosynthesis of pectic galactan, including the GALS1 galactan synthase and the UDP-galactose/UDP-rhamnose transporter URGT1. We have engineered plants with a more suitable biomass composition by applying these findings, in conjunction with synthetic biology and gene stacking tools. Plants were engineered to have up to fourfold more pectic galactan in stems by overexpressing GALS1, URGT1, and UGE2, a UDP-glucose epimerase. Furthermore, the increased galactan trait was engineered into plants that were already engineered to have low xylan content by restricting xylan biosynthesis to vessels where this polysaccharide is essential. Finally, the high galactan and low xylan traits were stacked with the low lignin trait obtained by expressing the QsuB gene encoding dehydroshikimate dehydratase in lignifying cells. In conclusion, the results show that approaches to increasing C6 sugar content, decreasing xylan, and reducing lignin content can be combined in an additive manner. Thus, the engineered lines obtained by this trait-stacking approach have substantially improved properties from the perspective of biofuel production, and they do not show any obvious negative growth effects. The approach used in this study can be readily transferred to bioenergy crop plants.

09 BIOMASS FUELS↗
DOE OSTI2019

Identification and manipulation of Neurospora crassa genes involved in sensitivity to furfural

Background: Biofuels derived from lignocellulosic biomass are a viable alternative to fossil fuels required for transportation. Following plant biomass pretreatment, the furan derivative furfural is present at concentrations which are inhibitory to yeasts. Detoxification of furfural is thus important for efficient fermentation. Here, we searched for new genetic attributes in the fungus Neurospora crassa that may be linked to furfural tolerance. The fact that furfural is involved in the natural process of sexual spore germination of N. crassa and that this fungus is highly amenable to genetic manipulations makes it a rational candidate for this study. Results: Both hypothesis-based and unbiased (random promotor mutagenesis) approaches were performed to identify N. crassa genes associated with the response to furfural. Changes in the transcriptional profile following exposure to furfural revealed that the affected processes were, overall, similar to those observed in Saccharomyces cerevisiae . N. crassa was more tolerant (by ~ 30%) to furfural when carboxymethyl cellulose was the main carbon source as opposed to sucrose, indicative of a link between carbohydrate metabolism and furfural tolerance. We also observed increased tolerance in a Δ cre-1 mutant (CRE-1 is a key transcription factor that regulates the ability of fungi to utilize non-preferred carbon sources). In addition, analysis of aldehyde dehydrogenase mutants showed that ahd-2 (NCU00378) was involved in tolerance to furfural as well as the predicted membrane transporter NCU05580 ( flr-1 ), a homolog of FLR1 in S. cerevisiae . Further to the rational screening, an unbiased approach revealed additional genes whose inactivation conferred increased tolerance to furfural: (i) NCU02488, which affected the abundance of the non-anchored cell wall protein NCW-1 (NCU05137), and (ii) the zinc finger protein NCU01407. Conclusions:We identified attributes in N. crassa associated with tolerance or degradation of furfural, using complementary research approaches. The manipulation of the genes involved in furan sensitivity can provide a means for improving the production of biofuel producing strains. Similar research approaches can be utilized in N. crassa and other filamentous fungi to identify additional attributes relevant to other furans or toxic chemicals.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Metabolic engineering of Zymomonas mobilis for anaerobic isobutanol production

Biofuels and value-added biochemicals derived from renewable biomass via biochemical conversion have attracted considerable attention to meet global sustainable energy and environmental goals. Isobutanol is a four-carbon alcohol with many advantages that make it attractive as a fossil-fuel alternative. Zymomonas mobilis is a highly efficient, anaerobic, ethanologenic bacterium making it a promising industrial platform for use in a biorefinery. In this study, the effect of isobutanol on Z. mobilis was investigated, and various isobutanol-producing recombinant strains were constructed. The results showed that the Z. mobilis parental strain was able to grow in the presence of isobutanol below 12 g/L while concentrations greater than 16 g/L inhibited cell growth. Integration of the heterologous gene encoding 2-ketoisovalerate decarboxylase such as kdcA from Lactococcus lactis is required for isobutanol production in Z. mobilis. Moreover, isobutanol production increased from nearly zero to 100-150 mg/L in recombinant strains containing the kdcA gene driven by the tetracycline-inducible promoter Ptet. In addition, we determined that overexpression of a heterologous als gene and two native genes (ilvC and ilvD) involved in valine metabolism in a recombinant Z. mobilis strain expressing kdcA can divert pyruvate from ethanol production to isobutanol biosynthesis. This engineering improved isobutanol production to above 1 g/L. Finally, recombinant strains containing both a synthetic operon, als-ilvC-ilvD, driven by Ptet and the kdcA gene driven by the constitutive strong promoter, Pgap, were determined to greatly enhance isobutanol production with a maximum titer about 4.0 g/L. Finally, isobutanol production was negatively affected by aeration with more isobutanol being produced in more poorly aerated flasks. This study demonstrated that overexpression of kdcA in combination with a synthetic heterologous operon, als-ilvC-ilvD, is crucial for diverting pyruvate from ethanol production for enhanced isobutanol biosynthesis. Moreover, this study also provides a strategy for harnessing the valine metabolic pathway for future production of other pyruvate-derived biochemicals in Z. mobilis.

09 BIOMASS FUELS↗