Laboratory-Scale Pretreatment and Hydrolysis Assay for Determination of Reactivity in Cellulosic Biomass Feedstocks
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Clostridium autoethanogenum is an acetogenic bacterium that autotrophically converts carbon monoxide (CO) and carbon dioxide (CO 2 ) gases into bioproducts and fuels via the Wood–Ljungdahl pathway (WLP). To facilitate overall carbon capture efficiency, the reaction stoichiometry requires supplementation of hydrogen at an increased ratio of H 2 :CO to maximize CO 2 utilization; however, the molecular details and thus the ability to understand the mechanism of this supplementation are largely unknown. In order to elucidate the microbial physiology and fermentation where at least 75% of the carbon in ethanol comes from CO 2 , we established controlled chemostats that facilitated a novel and high (11:1) H 2 :CO uptake ratio. We compared and contrasted proteomic and metabolomics profiles to replicate continuous stirred tank reactors (CSTRs) at the same growth rate from a lower (5:1) H 2 :CO condition where ~ 50% of the carbon in ethanol is derived from CO 2 . Our hypothesis was that major changes would be observed in the hydrogenases and/or redox-related proteins and the WLP to compensate for the elevated hydrogen feed gas. Our analyses did reveal protein abundance differences between the two conditions largely related to reduction–oxidation (redox) pathways and cofactor biosynthesis, but the changes were more minor than we would have expected. While the Wood–Ljungdahl pathway proteins remained consistent across the conditions, other post-translational regulatory processes, such as lysine-acetylation, were observed and appeared to be more important for fine-tuning this carbon metabolism pathway. Metabolomic analyses showed that the increase in H 2 :CO ratio drives the organism to higher carbon dioxide utilization resulting in lower carbon storages and accumulated fatty acid metabolite levels. This research delves into the intricate dynamics of carbon fixation in C. autoethanogenum, examining the influence of highly elevated H 2 :CO ratios on metabolic processes and product outcomes. The study underscores the significance of optimizing gas feed composition for enhanced industrial efficiency, shedding light on potential mechanisms, such as post-translational modifications (PTMs), to fine-tune enzymatic activities and improve desired product yields.
Sugarcane plant engineered to accumulate lipids in its vegetative tissue is being developed as a new bioenergy crop. The new crop would be a source of juice, oil, and cellulosic sugars. However, limited tolerance of industrially recognized yeasts towards inhibitors generated during the processing of lignocellulosic biomass to produce fermentable sugars is a major challenge in developing scalable processes for second-generation drop-in fuel production. To this end, hydrolysates generated from engineered sugarcane—‘oilcane’ bagasse contain added phenolics and fatty acids that further restrict the growth of fermenting microorganisms and necessitate nutrient supplementation and/or detoxification of hydrolysate which makes the fermentation process expensive. Herein, we propose a resourceful and economical approach for growing lab and commercial strains of S. cerevisiae on unrefined cellulosic sugars aerobically and fermentatively.
Clostridium phytofermentans was isolated from forest soil and is distinguished by its capacity to directly ferment plant cell wall polysaccharides into ethanol as the primary product, suggesting that it possesses unusual catabolic pathways. The objective of our present study was to understand the molecular mechanisms of biomass conversion to ethanol in a single organism, Clostridium phytofermentans, by analyzing its complete genome and transcriptome during growth on plant carbohydrates. The saccharolytic versatility of C. phytofermentans is reflected in a diversity of genes encoding ATP-binding cassette sugar transporters and glycoside hydrolases, many of which may have been acquired through horizontal gene transfer. These genes are frequently organized as operons that may be controlled individually by the many transcriptional regulators identified in the genome. Preferential ethanol production may be due to high levels of expression of multiple ethanol dehydrogenases and additional pathways maximizing ethanol yield. The genome also encodes three different proteinaceous bacterial microcompartments with the capacity to compartmentalize pathways that divert fermentation intermediates to various products. Lastly, these characteristics make C. phytofermentans an attractive resource for improving the efficiency and speed of biomass conversion to biofuels.
This project demonstrated microwave cracking of ethane with good product conversion and ethylene selectivity, with a short residence time (~0.001 sec). The laboratory scale equipment was designed and built, along with concept designs for larger scale implementation. The system was operated below atmospheric pressures, in the range of 15-55 torr, with argon as a carrier gas. The measured products included hydrogen, methane, acetylene, and ethylene. The results followed similar trends to those predicted by the modeling software SPYRO®, with the exception that the microwave appeared to produce slightly lower amounts of ethylene and methane, although enhanced analytical analysis should reduce the difference. Continued testing will be required to verify these results and quantify the energy consumption of microwave vs. conventional. The microwave cracking process is an attractive option due to the possibility of selectively heating the reaction volume rather than the reactor walls, which may allow novel reactor designs that result in more efficient production of ethylene. Supplemental studies are needed to continue the laboratory testing and refine processing parameters.
Our research focuses on transgenic strategies for modifying lignification to improve biomass quality, without leading to deleterious effects on plant performance. In order to accomplish this objective, we designed molecular strategies and selected appropriate transgenes for manipulating the expression of lignification-associated genes; we generated poplar engineered for altered lignin content and/or monomer composition, and field-tested them for fitness; we analyzed the impact of these transgenic strategies on metabolism in general and lignin biosynthesis in particular; and evaluated the ease with which cell wall deconstruction can be accomplished using both chemical and enzymatic means using wild-type and high syringyl poplar.
Idaho National Laboratory's Biomass Logistics Model has developed to a point for which a rigorous verification of the model and the results is appropriate. This verification will help add confidence to the model results and the system's design. The verification process consists of performing a number of predefined scenarios to exercise the most used and common paths and functions in the model. INL subject matter experts that understand the various components, but have not been actively involved in the development of the model, will review the results. This document is a summary of that activity.
Experiment results of EM separation show that the non-metallic inclusions were successfully pushed to the boundary layer of the crucible under EM force. Larger frequency and smaller current generate smaller thickness of accumulated inclusions. More detailed EM separation experiments are undergoing to investigate the factors that affect the removal efficient of inclusions from SoG-Si.
The goal of this proposed work is to develop and optimize the synthesis of mesoporous nanoparticle materials that are able to selectively sequester fatty acids from hexane extracts from algae, and to catalyze their transformation, as well as waste oils, into biodiesel. The project involves studies of the interactions between the functionalized MSN surface and the sequestering molecules. We investigate the mechanisms of selective extraction of fatty acids and conversion of triglycerides and fatty acids into biodiesel by the produced nanoparticles. This knowledge is used to further improve the properties of the mesoporous nanoparticle materials for both tasks. Furthermore, we investigate the strategies for scaling the synthesis of the catalytic nanomaterials up from the current pilot plant scale to industrial level, such that the biodiesel obtained with this technology can successfully compete with food crop-based biodiesel and petroleum diesel.
Air Products and Chemicals, Inc., is developing the sour pressure swing adsorption (PSA) technology which can be used to reject acid gas components (hydrogen sulfide [H 2 S] and carbon dioxide [H 2 S]) from sour syngas streams such as coal gasification syngas. In the current work, tests were conducted to investigate the impact of continuous exposure of real sour syngas and dilute levels of hydrochloric acid (HCl) and ammonia (NH 3 ) on the preferred adsorbent of that process. The results show a modest (~10%–15%) decrease in CO 2 adsorption capacity after sour syngas exposure, as well as deposition of metals from carbonyl decomposition. Continuous exposure to HCl and NH 3 yield a higher degree of CO 2 capacity degradation (up to 25%). These tests represent worst-case approaches since the exposure is continuous and the HCl and NH 3 levels are relatively high compare to an industrial sour syngas stream. Long-term PSA tests are needed to unequivocally evaluate the impact of cyclic exposure to these types of streams.