Silicon concentration and pH controls over competitive or simultaneous incorporation of iodate and chromate into calcium carbonate phases
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Two studies in the early 1980s described the leaching behavior of americium (Am) disposed as part of acidic high-salt processing wastes from the Hanford Site’s Plutonium Finishing Plant to nearby ground sediments. Here these batch leach experiments showed that the Am concentrations followed a linear log [Am] versus pH relationship with a slope of –1. Column leach experiments in the second study, however, did not follow this relationship and only ~30% of the americium desorbed even after extensive column leaching. Here, the 1980s research is re-examined along with previously unpublished information and, in light of recent published work, a plausible mechanism is proposed to explain these phenomena. Amorphous silica in the contaminated sediments is postulated to be the substrate responsible for both the exchangeable Am available for leaching and the retained low-leachable Am made evident in the column leach experiments. The exchangeable Am 3+ in the contaminated sediment leach experiments behaves with pH dependence similar to that observed for uptake onto amorphous silica of sodium (Na + ), calcium (Ca 2+ ), barium (Ba 2+ ), cadmium (Cd 2+ ), uranyl (UO 2 2+ ), ferric (Fe 3+ ), chromic (Cr 3+ ), cupric (Cu 2+ ), plumbous (Pb 2+ ), uranium(IV) (U 4+ ), plutonium(IV) (Pu 4+ ), zirconium (Zr 4+ ), analogue lanthanide (gadolinium, Gd 3+ , europium, Eu 3+ , and lutetium, Lu 3+ ) and curium (Cm 3+ ) ions as well as other studies with Am 3+ . Correspondingly, the residual low-leachable Am 3+ revealed in the column leach experiments is attributed to incorporation of Am 3+ within amorphous silica by dynamic Am 3+ sorption and silica precipitation processes.
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Hydrothermal pretreatment (HTP) using only water offers great potential to reduce the overall cost of the bioconversion process. However, traditional HTP performed in a batch has limitations in removing lignin and often needs to be performed under severe conditions to achieve reasonable pretreatment effects. Additionally, lignin left in the pretreated residue at these conditions is also highly condensed, thus possessing an even more adverse impact on the hydrolysis process, which requires high enzyme loadings. To address these technical challenges, HTP performed in a flow-through configuration was developed to simultaneously achieve near-complete hemicellulose recovery, high lignin removal and high sugar release. Despite facing challenges such as potentially large water usage, flow-through HTP still represents one of the most cost-effective and eco-friendly pretreatment methods. This review mainly covers the latest cutting-edge innovations of flow-through HTP along with structural and compositional changes of cellulose, hemicellulose, and lignin before and after pretreatment.
3-Hydroxypropionic acid (3-HP) and 1,3-propanediol (1,3-PDO) are value-added chemicals with versatile applications in the chemical, pharmaceutical, and food industries. Nevertheless, sustainable production of 3-HP and 1,3-PDO is often limited by the lack of efficient strains and suitable fermentation configurations. We report attempts have been made to improve the co-production of both metabolites through metabolic engineering of Escherichia coli and process optimization. First, the 3-HP and 1,3-PDO co-biosynthetic pathways were recruited and optimized in E. coli, followed by coupling the pathways to the transhydrogenase-mediated cofactor regeneration systems that increased cofactor availability and product synthesis. Next, pathway rebalancing and block of by-product formation significantly improved 3-HP and 1,3-PDO net titer. Subsequently, glycerol flux toward 3-HP and 1,3-PDO synthesis was maximized by removing metabolic repression and fine-tuning the glycerol oxidation pathway. Lastly, the combined fermentation process optimization and two-stage pH-controlled fed-batch fermentation co-produced 140.50 g/L 3-HP and 1,3-PDO, with 0.85 mol/mol net yield.
Anaerobic gut fungi (AGF) have potential to valorize lignocellulosic biomass owing to their diverse repertoire of carbohydrate-active enzymes (CAZymes). However, AGF metabolism is poorly understood, and no stable genetic tools are available to manipulate growth and metabolic flux to enhance production of specific targets, e.g., cells, CAZymes, or metabolites. Herein, a cost-effective, Arduino-based, continuous-flow anaerobic bioreactor with online optical density control is presented to probe metabolism and predictably tune fluxes in Caecomyces churrovis. Varying the C. churrovis turbidostat setpoint titer reliably controlled growth rate (from 0.04 to 0.20 h −1 ), metabolic flux, and production rates of acetate, formate, lactate, and ethanol. Bioreactor setpoints to maximize production of each product were identified, and all continuous production rates significantly exceed batch rates. Formate spike-ins increased lactate flux and decreased acetate, ethanol, and formate fluxes. The bioreactor and turbidostat culture schemes demonstrated here offer tools to tailor AGF fermentations to application-specific hydrolysate product profiles.
Despite its prominence, the ability to engineer Cupriavidus necator H16 for inorganic carbon uptake and fixation is underexplored. We tested the roles of endogenous and heterologous genes on C. necator inorganic carbon metabolism. Deletion of β-carbonic anhydrase can had the most deleterious effect on C. necator autotrophic growth. Replacement of this native uptake system with several classes of dissolved inorganic carbon (DIC) transporters from Cyanobacteria and chemolithoautotrophic bacteria recovered autotrophic growth and supported higher cell densities compared to wild-type (WT) C. necator in batch culture. Strains expressing Halothiobacillus neopolitanus DAB2 (hnDAB2) and diverse rubisco homologs grew in CO 2 similarly to the wild-type strain. Our experiments suggest that the primary role of carbonic anhydrase during autotrophic growth is to support anaplerotic metabolism, and an array of DIC transporters can complement this function. This work demonstrates flexibility in HCO 3 - uptake and CO 2 fixation in C. necator, providing new pathways for CO 2 -based biomanufacturing.
The production of cellulosic sugars is a pivotal strategy for advancing biomass bioconversion. This study evaluated the pretreatment of corn stover using ammonium sulfite and potassium hydroxide to develop comprehensive data on sugar, lignin, chemicals, and overall mass recovery profiles in a batch reactor at 80 °C. The results indicated significant improvements in delignification, deacetylation, enzymatic digestibility, and overall sugar yield. Specifically, pretreating corn stover with a solution of 40 wt% potassium hydroxide and 15 wt% ammonium sulfite at 80 °C for 2 h achieved 78.9 % lignin removal and 82.1 % acetyl removal, resulting in a total sugar yield exceeding 87.5 % with an enzyme loading of 12.5 mg protein/g-glucan plus xylan. The pretreated spent liquor, containing ammonium, potassium, sulfur, biomass-derived organics, and inorganics, demonstrated substantial potential as a fertilizer. The techno-economic analysis projected a minimum sugar selling price of $0.285 per pound, supporting the ongoing development and implementation of chemical-recovery-free pretreatment technology.
Rhodosporidium toruloides has gained increasing interests as a promising non-model host organism to produce a wide range of bioproducts from lignocellulosic biomass. Increasing the bioproduct titers, rates, and yields remains a challenge, largely due to a lack of robust and well-characterized genetic tools in this host. Here we developed an inducible flippase (FLP) and flippase recognition target (FRT) system that enables genetic manipulations without the need for additional selection markers. Synthetic inducible promoters were established, enabling regulation of FLP expression and efficient antibiotic marker removal. Leveraging this system, we engineered a strain to optimize 3-hydroxypropionic acid (3HP) production. Over four rounds of iterative genomic editing to resolve pathway bottlenecks, we achieved a 3HP titer of 69.4 g/L in fed-batch fermentation - the highest level reported in yeast from lignocellulosic hydrolysates. The engineered high 3HP producing strain offers a robust platform for sustainable bio-based chemical production from lignocellulosic feedstocks.
Efficient and cost-competitive bioproduction calls for utilizing CO2-derived feedstocks, such as products from electro-reduction of CO2 and hydrolysate from lignocellulosic biomass. However, efficiently using all their carbon components, including acetate, glucose, and xylose, remains a challenge. Here, we characterize Lipomyces tetrasporus, a novel, robust yeast strain capable of effectively assimilating these carbon sources. We used an integrated systems biology approach combining ¹³C metabolic flux analysis, dynamic labeling experiments, and RNA sequencing. We conducted the first metabolic flux analysis for glucose, xylose, and acetate catabolism in this species. Dynamic labeling revealed a highly active TCA cycle during acetate metabolism, evidenced by rapid citrate and malate accumulation. The strain demonstrated strong NADH/NADPH production and acetyl-CoA synthase activity. Using insights and gene targets from this analysis, we engineered L. tetrasporus for malate production. The engineered strain produced 7.5 g/L malic acid (0.25 g/g yield) in shake flasks with glucose-acetate media and 28.8 g/L malic acid at a yield of 0.20 g/g in fed-batch mode with corn-stover hydrolysate. Together, these insights and rational strain engineering establish L. tetrasporus as a versatile, Crabtree-negative platform that is an energy-CO2-bioproduction nexus for channeling CO2 carbon into value-added bioproducts.
Machine learning-based data-driven modeling can allow computationally efficient time-dependent solutions of PDEs, such as those that describe subsurface multiphysical problems. In this work, our previous approach (Kadeethum et al., 2021d) of conditional generative adversarial networks (cGAN) developed for the solution of steady-state problems involving highly heterogeneous material properties is extended to time-dependent problems by adopting the concept of continuous cGAN (CcGAN). The CcGAN that can condition continuous variables is developed to incorporate the time domain through either element-wise addition or conditional batch normalization. Moreover, this framework can handle training data that contain different timestamps and then predict timestamps that do not exist in the training data. As a numerical example, the transient response of the coupled poroelastic process is studied in two different permeability fields: Zinn & Harvey transformation and a bimodal transformation. The proposed CcGAN uses heterogeneous permeability fields as input parameters while pressure and displacement fields over time are model output. Our results show that the model provides sufficient accuracy with computational speed-up. This robust framework will enable us to perform real-time reservoir management and robust uncertainty quantification in poroelastic problems.
Carbon fiber is a highly desired material for structural applications requiring high strength and stiffness and low weight but has seen only incremental improvements in properties over the last few decades. Further increases in carbon fiber specific properties, including specific strength and specific modulus, would further propel its unique capabilities. One method to produce high specific property carbon fibers for structural applications is the development of hollow carbon fibers. In this work, we report on the early development of polyacrylonitrile-derived structural hollow carbon fibers. Here, multifilament, continuous tow, polyacrylonitrile-based precursor hollow fibers were successfully produced utilizing a segmented arc spinneret approach. When batch oxidized, the hollow precursor fibers demonstrated evidence of oxidation proceeding from both the interior and exterior of the filament. Further results suggested that reducing the precursor hollow fiber wall thickness would allow for complete, homogeneous oxidation, thereby avoiding the skin-core structure often observed in commercial carbon fiber. Here, hollow carbon fibers were as small as 35 µm outer diameter, 22 µm inner diameter (6.5 µm wall thickness). At these diameters, the hollow carbon effective fiber specific strength was 0.54 N/tex and the effective specific modulus was 120 N/tex, approaching the effective specific modulus of T700S at 136 N/tex.
We report scalable production of carbon nanotubes (CNTs) requires catalysts and reaction conditions that provide high nucleation efficiency. In situ characterization methods such as environmental transmission electron microscopy (ETEM) can reveal fundamental mechanisms of synthesis, but to date have primarily provided qualitative observations on small sample sizes. Here, quantitative analysis is performed using high-resolution, high-rate video capture of ETEM experimentation coupled with automated image processing, involving computer vision algorithms and convolutional neural networks. By this approach, we detect distinct nanoparticle formation from an alumina-supported iron thin film and subsequent CNT nucleation from the nanoparticles. The statistical summary of particles in each video shows that, compared to a H 2 -only atmosphere, pretreatment of the catalyst with carbon added to the H 2 atmosphere results in a smaller average particle diameter, a 2-fold increase in particle density (to 5300 particles/μm 2 ), a 3-fold increase in CNT nucleation efficiency (to 92%), and more than a 5-fold increase in CNT density (to 4800/μm 2 ). Addition of carbon during exposure to H 2 is also more effective than NH 3 at dewetting the catalyst film and increasing the CNT nucleation efficiency, in spite of NH 3 being a stronger reducing agent for iron. Insights from this study are applicable to improving CNT yield and productivity in both batch-style and continuous processes.
Owing to their remarkable properties and ordered architecture, vertically aligned CNT forests (VACNT) hold promises for countless applications ranging from energy storage to multifunctional fiber production. Over the last three decades, studies of carbon nanotube (CNT) growth have provided invaluable insight into the CNT nucleation and growth mechanism and enabled critical advancements in nanotechnology. Nevertheless, the ability to fully harness the exceptional VACNT properties in commercial devices is bottlenecked by compatibility of current CNT synthesis processes with large-scale manufacturing. Metal foil substrates present an economic alternative to traditional insulating substrates, like silicon or quartz, and are compatible with both roll-to-roll and automated, large-scale, batch processes. Compared to CNT growth on Si substrates, metal foils present additional challenges, such as increased roughness and reactivity at high temperatures. Nonetheless, many successful VACNT growths on conducting metal foils have been reported, which demonstrate the feasibility of metal substrate use for large-volume production of high-quality CNTs. Here, in this review, we discuss advancements in the field of VACNT growth on metal foil and specifically examine different choices for metal substrates, barrier and catalyst layers, deposition and growth methods, resulting CNT characteristics, and highlight a number of applications that benefit from CNT growth on metal substrates and transition to large-scale manufacturing.
The presence of homoacetogenesis and hydrogen production-consumption loop significantly hinders the efficient hydrogen recovery in single chamber microbial electrolysis cells (MECs). Furthermore, no effective inhibition method has been proposed against homoacetogens hitherto. In this study, we demonstrated the effectiveness of chloroform as a homoacetogen inhibitor in MECs. In small MECs (320 ml) operated in batch mode, chloroform with concentrations of 0.02% (v/v) and 0.03% (v/v) ceased homoacetogenesis with nonfermentable substrate (acetate), enhancing hydrogen yield and cathodic hydrogen recovery from 21% and 14% to 94% and 90%, respectively. Electrochemical performance was not significantly affected as current density (based on anode surface area) only decreased from 18 A/m 2 to 13 A/m 2 with chloroform concentration increased from 0% (v/v) to 0.03% (v/v). However, when using a fermentable substrate (glucose), the inhibition against homoacetogens by 0.01% (v/v) chloroform led to a significant decrease in current density possibly due to the decreased production of acetate, which reduced the substrate concentration for exoelectrogenesis. Nevertheless, in a larger MEC (10 L), using an immobilized sludge culture and higher glucose concentration (56mM), 0.02% (v/v) chloroform enhanced hydrogen production rate from 0 L/L/D to 4.9 L/L/D and current density from 12-16 A/m 2 to 18-21 A/m 2 . Microbial community analysis revealed that the amount of homoacetogenic Acetobacterium spp. was eliminated in the cathodic biofilms and planktonic cells by chloroform. Future improvement of this method, through reducing the residual chloroform concentration and developing more environment friendly inhibitor using similar mechanisms, may lead to the practical application in MECs.
The deacetylation process is a key step in the lignocellulosic biomass pretreatment process that currently is done in batch reactors. A flow-through diffuser type reactor is a promising approach to make the process more efficient and economically favorable. A washing step before deacetylation can further reduce chemical usage and increase the process efficiency. However, the highly compacted biomass particles can cause significant pressure-drop for fluid flow through the bed, which in the worst case can cause self-enhancing pressure-drop leading to reactor blocking. Detailed understanding of the fundamentals of flow through a compressible packed bed is necessary to scale up the washing and deacetylation process. This work closely examines the classical Kozeny Carman equation and proposes modifications to accurately predict the pressure-drop at different compaction states. Here, a compressible packed bed model is further developed based on the interaction between solid mechanics and fluid dynamics. Experimental measurements of water flow through a packed bed of corn stover show good agreement with the developed model. Corn stover washing process scale up simulations show that smaller reactor diameter, higher friction lining material, lower process throughput, and higher process temperature is favorable in stabilizing the bed from extensive compression when the bed height is fixed.
Here we conduct an experimental study of fructose dehydration to 5-hydroxymethyl furfural (HMF) in a biphasic microreactor as a function of residence time, temperature, and sugar loading using methyl butyl iso-ketone (MIBK) and 2-pentanol as extracting solvents. We demonstrate stable operation with maximum HMF yields of 93% and 87% in the two solvents, respectively, at 200 °C and a residence time of 2 s for a 5 wt% fructose aqueous feed. We report the highest optimal HMF space–time yield of 60 kg/L-hr at 200 °C (10 – 2,500-fold higher than published reports). Unexpectedly, an optimum organic-to-water ratio exists that depends on the solvent. Notably, we observe experimentally an increased fructose rate and HMF yield well above the extraction thermodynamic limit and hypothesize that the solvent plays a dual role, that of an extractant to protect HMF from degradation and a modifier of the fundamental chemistry. We expose mass transfer limitations of microreactors at longer residence times and higher temperatures and provide reactivity maps for their design. We show that batch reactors are unfit for high throughput and distributed manufacturing where small, farm-based systems are necessary.
Rising concerns for sustainability and global climate change have driven the development of sustainable pro- duction pathways for biofuels and chemicals from lignocellulosic biomass via integrated biological and chemical processes. We constructed an engineered Saccharomyces cerevisiae capable of producing 2,3-butanediol (2,3-BDO) from glucose without accumulating ethanol and glycerol, which hinder downstream processing of 2,3-BDO, through extensive metabolic reprogramming. Specifically, we introduced heterologous 2,3-BDO biosynthetic enzymes and deleted the major isozymes of ethanol and glycerol biosynthetic enzymes. In addition, we introduced an NAD + regenerating Pyruvate-Malate (PM) cycle and enhanced the NAD + regenerating capability of the PM cycle to resolve the redox imbalance from the deletion of ethanol and glycerol production pathways. The resulting engineered yeast produced 109.9 g/L of 2,3-BDO with a productivity of 1.0 g/L/h and a yield of 0.36 g/ g glucose in a fed-batch fermentation. We also conducted techno-economic analysis (TEA) and life cycle assessment (LCA) of the production of methyl ethyl ketone (MEK) through catalytic dehydration of 2,3-BDO. A TEA based on the experimental results indicated that the minimum product selling price (MPSP) was estimated to be $1.90/kg. Regarding cradle-to-grave LCA, 100-year global warming potential (GWP100) and fossil energy consumption (FEC) were found to be 0.37 kg CO 2 eq/kg and 3.1 MJ/kg, respectively. These results demonstrated the feasibility of cost-competitive and sustainable bio-based MEK production via yeast fermentation. In addition, we explored the possibility of using the fermentation broth containing 2,3-BDO as a biostimulant inducing drought tolerance in plants. As a result, the yeast 2,3-BDO fermentation broth can induce drought tolerance in Arabidopsis thaliana without a complicated purification process.