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A novel open-source cultivation system helps establish the first full cycle chemosynthetic symbiosis model system involving the giant ciliate Zoothamnium niveum

Symbiotic interactions drive species evolution, with nutritional symbioses playing vital roles across ecosystems. Chemosynthetic symbioses are globally distributed and ecologically significant, yet the lack of model systems has hindered research progress. The giant ciliate Zoothamnium niveum and its sulfur-oxidizing symbionts represent the only known chemosynthetic symbiosis with a short life span that has been transiently cultivated in the laboratory. While it is experimentally tractable and presents a promising model system, it currently lacks an open-source, simple, and standardized cultivation setup. Following the FABricated Ecosystems (EcoFABs) model, we leveraged 3D printing and polydimethylsiloxane (PDMS) casting to develop simple flow-through cultivation chambers that can be produced and adopted by any laboratory. The streamlined manufacturing process reduces production time by 86% and cuts cost by tenfold compared to the previous system. Benchmarking using previously established optimal growth conditions, the new open-source cultivation system proves stable, efficient, more autonomous, and promotes a more prolific growth of the symbiosis. For the first time, starting from single cells, we successfully cultivated the symbiosis in flow-through chambers for 20 days, spanning multiple generations of colonies that remained symbiotic. They were transferred from chamber to chamber enabling long-term cultivation and eliminating the need for continuous field sampling. The chambers, optimized for live imaging, allowed detailed observation of the synchronized growth between the host and symbiont. Highlighting the benefit of this new system, we here describe a new step in the first hours of development where the host pauses growth, expels a coat, before resuming growth, hinting at a putative symbiont selection mechanism early in the colony life cycle. With this simple, open-source, cultivation setup, Z. niveum holds promises for comparative studies, standardization of research and wide adoption by the symbiosis research community.

59 BASIC BIOLOGICAL SCIENCES

Degradable Biocomposite Thermoplastic Polyurethanes

In this project, the team developed tough and degradable biocomposite thermoplastic polyurethanes (TPUs) by incorporating bacterial spores into TPUs as a biofunctional living filler. The team screened various bacteria and selected the Bacillus subtilis ATCC 6633 strain as the final candidate, primarily due to its genomic availability, sporulation ability and TPU assimilation activity. The heat-shock tolerance of ATCC 6633 spores was further improved through evolutionary engineering via Adaptive Laboratory Evolution (ALE), demonstrating a 17.7-fold enhanced germination efficiency post heat-shock treatment compared to the wild-type strain (WT). The team fabricated biocomposite TPUs by incorporating lyophilized powder of heat-shock tolerized (HST) spores during the hot melt extrusion (HME) of TPU at 135 °C. The baseline TPU used in this project is a commercially available soft-grade TPU (BCF45) manufactured by BASF. Colony forming unit (CFU) assays quantified that WT and HST spores in the TPU matrix retained approximately 20% and 100% survivability, respectively, after HME. Tensile testing demonstrated that the spores behaved as a polymer-reinforcing filler, positively affecting the overall tensile properties of the biocomposite TPU. For example, biocomposite TPU with WT and HST spores (BC TPU WT and BC TPU HST , respectively) exhibited up to 25% and 37% improved toughness, respectively, compared to TPU without spores. BC TPU HST showed remarkably improved disintegration in autoclaved compost (92% mass loss in 5 months), which simulated a microbially poor environment for TPU degradation. When compared to TPU without spores (44% mass loss in 5 months) the acceleration of degradation is marked. Respirometry confirmed that 72% of BC TPU HST was biomineralized into CO2 within 6 months, indicating that spores in the biocomposite TPU were germinated by utilizing nutrients in the autoclaved compost, facilitating TPU degradation at the end of the material's life. The team demonstrated the scale-up of biocomposite TPU fabrication using continuous extrusion and injection molding techniques. Processing conditions optimized in a lab-scale microcompounder were successfully transferred to a continuous extruder with a 30-fold increased throughput. Biocomposite TPUs prepared using these industry-relevant processes showed comparable toughness improvements to samples prepared in the lab-scale extruder. Excitingly, following compounding in the pilot-extruder the composite material could be injection molded, while retaining high spore viability and similar toughness improvements. The team also found that spores in biocomposite TPU served as antioxidants, preventing toughness decay during the recycled extrusion of BC TPU HST . Long-term storage tests over one year showed that the addition of spores had no negative effect on the longevity of the TPU. Furthermore, the team demonstrated the fabrication of spore-bearing biocomposite polymers with other polyesters such as PBAT, PLA, and PCL. We obtained promising preliminary data that showed overall toughness improvements for all polymers with spore addition. Finally, life cycle assessment (LCA) and techno-economic analysis (TEA) were carried out, which indicated minimal additional cost of fabrication. Overall, a tough and degradable biocomposite thermoplastic was successfully developed through this project, with all tasks completed successfully, achieving >100% of the objectives.

36 MATERIALS SCIENCE

Distributed Quantum-Enhanced Optimization: A Topographical Preconditioning Approach for High-Dimensional Search

Optimization problems become fundamentally challenging as the number of variables increases. Because the volume of the search space grows exponentially, classical algorithms frequently fail to locate the global minimum of non-convex functions. While quantum optimization offers a potential alternative, mapping continuous problems onto near-term quantum hardware introduces severe scaling limits and barren plateaus. To bridge this gap, we propose the Distributed Quantum-Enhanced Optimization (D-QEO) framework. Instead of forcing the quantum processor to find the exact minimum, we use it simply as a topographical preconditioner. The QPU maps the landscape to locate the most promising basin of attraction, generating high-quality seed points for a classical GPU-accelerated solver to refine. To make this approach viable for utility-scale problems, we exploit the mathematical structure of separable functions. This allows us to cut a 50-qubit (i.e., $2^{50}$) global search space into independent and manageable sub-spaces using 5-qubit subcircuits. By executing these fragments concurrently with CUDA-Q, we completely bypass the overhead of cross-register entanglement and classical tensor knitting for separable functions. Benchmarks on the 10-dimensional Rastrigin and Ackley functions show that D-QEO prevents the exponential failure rates observed in purely classical algorithms. Furthermore, this quantum warm-start significantly reduces the number of classical BFGS iterations required to converge, providing a highly practical blueprint for utilizing near-term quantum resources in complex global search.

Soos, Dominik [Old Dominion U.]

An optimization approach for cryogenic distillation-based protium removal systems in magnetic fusion energy fuel cycles

Tritium inventory reduction for fusion power plants is central to the successful adoption of fusion energy. The advent of direct internal recycling in deuterium-tritium fusion fuel cycle design has led to significant reduction in startup tritium inventory estimates for fusion power plants but requires an isotope rebalancing and protium removal (IRPR) system to ensure acceptable isotopic fuel composition. Cryogenic distillation is a potential solution for near-term deployment in an IRPR system due to its demonstrated performance in similar operating regimes. Using protium removal in the fuel cycle as the primary performance metric, this paper presents an optimization methodology for a single-column continuous cryogenic distillation-based IRPR system for a 500 MW fus magnetic fusion device. Distillation column optimization was performed using the CryOgenic Distillation For Isotopic Separation of Hydrogen (CODFISH) code developed at Savannah River National Laboratory. The distillation column design presented maximizes direct recycling of hydrogen isotopes from the fusion chamber exhaust to the fueling system while minimizing IRPR system steady-state tritium inventory. The optimized IRPR distillation column presented achieves direct recycling of 60 % of the hydrogen isotopes in the fusion chamber exhaust with an estimated steady-state system tritium inventory <30 g. In conclusion, the optimized IRPR distillation column operation was then used to estimate the design requirements for a detritation column to treat the IRPR system effluent stream.

CryOgenic distillation

Complete Demonstration of a Prototype Version of FORCE User Interface and Conduct Analyst Survey Collecting Feedback on Interface Features and Usability

In 2024 the US Department of Energy (DOE) Office of Nuclear Energy (NE) Integrated Energy System (IES) program continued to develop the Framework for Optimization of Resources and Economics (FORCE) analysis ecosystem into a more traditional toolset with simplified software installation, automated workflows, and interactive results visualization. The DOE-NE Nuclear Energy Advanced Modeling and Simulation (NEAMS) Workbench continued to be leveraged for user input, application workflow and runtime environment, and interactive results visualization capabilities. This report documents the demonstration of a FORCE User Interface (UI) prototype and the results of a survey of analysts’ using the Holistic Energy Resource Optimization Network (HERON) tool in FORCE with the prototype UI.

97 MATHEMATICS AND COMPUTING

In-situ sensor monitoring of multi-class gas porosity formation in laser powder bed fusion using convolutional neural network

In-situ monitoring of defect formation remains a significant challenge in the laser powder bed fusion (LPBF) process. Recent advances have enabled real-time defect detection with machine learning and in-situ sensing technologies; however, most studies focus on binary classification of keyhole pores, limiting nuanced multi-class pore differentiation and formation mechanisms. This work introduces a multi-class pore detection framework (no pore, small pores < 15 µm, and large pores > 15 µm) by leveraging photodiode sensor data alongside high-fidelity synchrotron X-ray imaging. The 15 µm threshold is selected to distinguish between two fundamentally different defect mechanisms, following the physical size-mechanism boundary established by prior high-resolution synchrotron X-ray characterization of Al6061 LPBF. Distinguishing these classes is critical because large keyhole pores are structurally detrimental, whereas small gas pores are often benign, requiring different process control strategies. Thermal emission monitoring data collected simultaneously with high-speed X-ray imaging at the Stanford Synchrotron Radiation Lightsource (SSRL), are correlated with subsurface melt pool dynamics to establish ground truth. Continuous Wavelet Transform (CWT) with optimized parameters converts the photodiode time-series signals into time–frequency images, facilitating feature extraction. Convolutional Neural Networks (CNN) are then applied for real-time multi-class pore classification in an average inference time of 1 ms per signal window. It achieves 79% accuracy and an Area Under the Receiver Operating Characteristic curve (AUC ROC) score of 0.89 with five-fold cross-validation. The results demonstrate that coupling CWT-based feature engineering with CNN architecture enables reliable multi-class pore detection in Al6061 builds using affordable in-situ sensors. This approach advances scalable and affordable quality assurance in additive manufacturing by moving beyond binary defect detection toward more nuanced classification of porosity mechanisms with in-situ sensors and machine learning.

Laser powder bed fusion, Multi-class pores, In-sit

Development and assessment of hierarchical multi-reward reinforcement learning based potential for silicene with state-of-the-art models

We develop a new interatomic force field for Silicene, a 2D material with a buckled hexagonal lattice structure with high polymorphism. We introduce new parameterizations of a Tersoff model using a hierarchical multi-reward reinforcement learning (RL) methodology coupled with a continuous Monte Carlo Tree Search optimization. Our model significantly outperforms existing methods by enhancing the accuracy of predictions for the structural and thermodynamic properties of seven silicene polymorphs-including structure, energy, equation of state, elasticity, and phonon dispersion-when compared to established models. We further make a comprehensive comparison of the various models in predicting the mechanical and thermal properties of silicene. We trace the origin of the improved performance to the description of the angular dependence in the bond-order term, suggesting that modifying the angular terms in short-range models is essential to capture the structural diversity in low dimensional systems.

2D materials

Constraining hydrodynamic models of inertial confinement fusion implosions using capsule surrogate experiments

We conduct capsule surrogate experiments at the National Ignition Facility to calibrate radiation hydrodynamic simulations to infer hydrodynamic conditions that are not observable in indirect drive ignition implosions. We tune the simulations by applying laser power and cross beam energy transfer (CBET) saturation multipliers to match the observables from capsule surrogate experiments. Shock timing, velocity, and symmetry are measured in liquid D 2 filled Keyhole capsule surrogate experiments and implosion trajectory, stagnation time, and shape time history are measured in in-flight 2D backlit x-ray radiography experiments (“2DConA”) of D 2 gas filled capsule implosions. Calibrated simulations suggest that the N210808 ignition implosion (fusion target gain = 0.7) had a shell mass remaining at stagnation of less than the nominal %5 (3.8%) and resulted in less confinement. For N221204, the shell was made 5.75 μm thicker to trade implosion velocity for increased confinement and resulted in a target gain = 1.5 with a shell mass remaining of 5.7%. Furthermore, a single adjusted model can reproduce all shock timing data as changes are made to shell thickness (79–85 μm) and laser wavelength separation (1.8–4.0 Å). However, for the 2DConA implosions, a 5% variation in the peak power laser multipliers and a 30% variation in late-time CBET between experiments are needed to match the observed stagnation times, in-flight $P_2$ shape, and hot-spot $P_2$ shape. While progress is being made to improve the models in simulations using focused experiments, capsule surrogate experiments will continue to be needed to optimize future ignition designs.

Lasers

Multilayer Architecture Barriers for High Temperature Nuclear Applications

Innovative thin film multilayer metal/ceramic barrier coatings have been developed by Argonne National Laboratory to protect fuel cladding against different phenomena that can be encountered under the harsh operating conditions of advanced nuclear reactors. One phenomenon is Fuel-Cladding Chemical Interaction (FCCI), which can represent a challenge to the performance of steel-clad metallic fuels in sodium-cooled fast reactors (SFRs), especially when operating to high burnups at elevated cladding temperatures, and long fuel residence time. Another phenomenon is the corrosion of zirconium-based cladding under prototypic boiling water reactor (BWR) conditions and associated cladding during off normal conditions, including loss of coolant accidents (LOCA). This work provides details of the development and testing of a thin film (few microns) multilayer metal/ceramic (CrY/Y 2 O 3 ) coatings on steel cladding and its performance against FCCI. The coating architecture evolved through systematic mechanical testing, radiation tolerance assessments, and high-temperature diffusion studies from a preliminary multilayer concept to an optimized high-performance design. The optimized structure leverages stoichiometric Y 2 O 3 for its exceptional radiation stability, chemical inertness, and ultra-low fission product diffusivity (e.g., Ce diffusivity ~10 -20 m 2 /s in irradiated Y 2 O 3 at 650 °C). This is paired with ductile chromium-rich interlayers (Cr 95 Y 5 ), which significantly enhance fracture toughness and mechanical resilience. Design refinements, including increased ceramic layer thickness and optimized Cr–Y interlayer composition, eliminated fracture propagation and preserved coating integrity under extreme mechanical stress (>10× typical reactor conditions) and prolonged radiation exposure (~300 dpa). Those conditions are well beyond expected conditions during normal operations in SFR, and cover some of the transient conditions as well. In addition, the coating deposition was demonstrated to be applied uniformly to the inner surface of steel tubes representing prototypic SFR cladding. Another phenomenon under consideration in this work is the corrosion of zirconium-based cladding under prototypic boiling water reactor (BWR) conditions and associated off normal conditions including loss of coolant accident (LOCA). Another multilayer thin film coating is considered for mitigation of this phenomenon, where a coating of yttrium aluminum and yttrium aluminum ceramic/metal thin film coating. Preliminary results on the coating performance during high temperature thermal cycling (up to 1000 C) have shown promising performance of the coating. On-going optimization of the coating will continue in FY26.

22 GENERAL STUDIES OF NUCLEAR REACTORS

60 Co γ Irradiation of a CMS CROC v2 to 1.6 Grad

This paper reports the irradiation of the production version of the readout chip for the CMS HL-LHC pixel detector, CROC~v2, to 1.6~Grad using γ s from a 60 Co source at Sandia National Laboratory. The CROC~v2 was kept powered, configured, and maintained as close as possible to its nominal operating temperature of -20~ o C during irradiation. Internal chip voltages, pixel response, and diagnostic ring oscillators were continuously monitored, and thresholds were optimized frequently. The CROC~v2 performed according to specifications at 1.6~Grad. Where available, these results will be compared with those for this and earlier iterations of the RD53 readout chip irradiated to a few Grad with lower-energy X-ray sources.

72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS

Investigation of Debonding Effect in Internal Replacement Pipe System Under Lateral Loading

The aging pipeline infrastructure around the world necessitates immediate rehabilitation. Internal replacement pipe (IRP) is a trenchless system offering a versatile and cost-effective solution across a variety of industries, including oil, natural gas, water, and wastewater. As a structural pipeline repair system, IRPs are subject to lateral deformation because of surface traffic loading. The present study evaluates the impact of adhesion between the host pipe and the IRP, with a focus on assessing the debonding effect on the behavior of the repair system under lateral deformation and bending. This was achieved using a comprehensive approach, including experimental, numerical, and analytical techniques. Varying levels of adhesive strength resulting from different methods of surface preparation were considered. The effectiveness of the IRP system on both discontinuous host pipes with various crack widths and continuous host pipes was also investigated. The results demonstrate that adhesive strength exerts a significant influence on the repair system, especially in the case of narrow circumferential cracks, while its impact on the continuous system is minimal. For optimal performance, it is essential to choose adhesives that possess sufficient shear strength while also accounting for the required debonding length. This approach ensures that minor discontinuities are effectively controlled, thereby enhancing the system′s fatigue life. The reliable determination of the maximum allowable shear strength for the adhesive or the debonding length can ensure that it does not negatively affect fatigue life. The findings presented in this study offer new insights into the development of trenchless repair techniques that can enhance system performance and extend service life.

Tien, Tri C. M.

Explainable physics-based constraints on reinforcement learning for accelerator optimization

We present a reinforcement learning (RL) framework for optimizing particle accelerator experiments that builds explainable physics-based constraints on agent behavior. The goal is to increase transparency and trust by letting users verify that the agent’s decision-making process incorporates suitable physics. Our algorithm uses a learnable surrogate function for physical observables, such as energy, and uses them to fine-tune how actions are chosen. This surrogate can be represented by a neural network or by an interpretable sparse dictionary model. We test our algorithm on a range of particle accelerator optimization environments designed to emulate the Continuous Electron Beam Accelerator Facility at Jefferson Lab. By examining the mathematical form of the learned constraint function, we are able to confirm the agent has learned to use the established physics of each environment. In addition, we find that the introduction of a physics-based surrogate enables our RL algorithms to reliably converge for difficult high-dimensional accelerator optimization environments.

explainability

Binary Quantum Control Optimization with Uncertain Hamiltonians

Optimizing the controls of quantum systems plays a crucial role in advancing quantum technologies. The time-varying noises in quantum systems and the widespread use of inhomogeneous quantum ensembles raise the need for high-quality quantum controls under uncertainties. In this paper, we consider a stochastic discrete optimization formulation of a discretized binary optimal quantum control problem involving Hamiltonians with predictable uncertainties. We propose a sample-based reformulation that optimizes both risk-neutral and risk-averse measurements of control policies, and solve these with two gradient-based algorithms using sum-up-rounding approaches. Furthermore, we discuss the differentiability of the objective function and prove upper bounds of the gaps between the optimal solutions to binary control problems and their continuous relaxations. We conduct numerical simulations on various sized problem instances based on two applications of quantum pulse optimization; we evaluate different strategies to mitigate the impact of uncertainties in quantum systems. In conclusion, we demonstrate that the controls of our stochastic optimization model achieve significantly higher quality and robustness compared with the controls of a deterministic model.

conditional value-at-risk (CVaR)

Numerical Modeling & Optimization of the iProTech Pitching Inertial Pump (PIP) Wave Energy Converter (WEC) (CRADA Final Report)

This project represents a continuation of the collaboration between iProTech and NLR to simulate, optimize and design the iProTech Pitching Inertial Pump (PIP) device. The objectives of this TEAMER project are twofold: 1. Refining the physical characteristics of the existing iProTech PIP WEC-Sim model to enhance the model’s fidelity and include controllable components. Key model enhancements target the inclusion of Coulomb friction, the introduction of a controllable bypass valve, and the replacement of traditional check valves with advanced motorized ones. 2. Exploring traditional and advanced control algorithms. From traditional methods like latching control to cutting-edge reinforcement learning (RL) algorithms, the goal is to ensure the PIP device's adaptability and optimal performance across a range of ocean conditions. NLR is tasked with augmenting the WEC-Sim model and implementing the control algorithms, culminating in performance comparison analyses. iProTech will update their existing 3D models, advise on model improvements, and determine crucial system metrics. WEC-Sim, developed in MATLAB/SIMULINK with Simscape Multibody, is the main piece of software that will be used in this project. Coupled with the MATLAB RL Toolbox, it offers a robust platform for in-depth simulation and optimization of the iProTech PIP device. Building on previous work to explore the PIP design space and optimize its geometry, mass distribution, center of gravity and other key parameters, this project aims to refine iProTech’s existing numerical models and develop effective control algorithms that can seamlessly integrate into their future hardware testing campaigns.

16 TIDAL AND WAVE POWER

Net Present Value Optimization of a Natural Gas Combined Cycle Plant with CO 2 Capture using a Water-Lean Solvent Considering Transient Electricity Price for Multiple Regions

Global CO 2 emissions are increasing at about a 1.5% rate per year. Fossil fuel-based plants are one of the main contributors to this rise. In the power generation industry, fossil fuel plants are dominant, and many plants are under development. In this study, a natural gas combined cycle (NGCC) power plant with postcombustion capture using a leading water-lean solvent is considered. For optimal design and operating schedule, large-scale dynamic optimization is undertaken for net present value (NPV) optimization. The first principle dynamic model of NGCC is developed, including a model of the highly efficient H-class gas turbines. For computational tractability of the dynamic optimization problem, a reduced-order model is developed by using the Hankel singular value decomposition. A waterlean solvent, N-(2-ethoxyethyl)-3-morpholinopropan-1-amine, is used for carbon capture. A model of the capture system is developed in Aspen Plus, which is used to develop a reduced-order model by using ALAMO, a machine learning software. In addition, a reduced model of the CO 2 compression system with a dehydration unit is also considered. The integrated system is used for NPV optimization by using the Python-based PYOMO platform. The PCC process is analyzed for three configurations-conventional packed bed, rotating packed bed (RPB), and a combination of RPB and direct contact cooler. The NPV optimization is performed for 14 regional markets by considering year-long clustered and continuous locational marginal price data with a 1 h interval. Optimization results show that the PCC can achieve 90% CO 2 capture with a positive NPV for six regions. Sensitivity studies conducted by using the PCC configurations indicate that the process is economically feasible for 9 regions out of 14 regional electricity markets with NPV values in the range of 33−540 $MM.

cabon capture

Adaptive PID Gain Scheduling Control for Hydropower Turbine Using Neural CDE and Stochastic Distribution Shaping

This paper introduces a gain-scheduling PID controller design strategy for hydroturbine frequency control mode. This scheme first uses real data to learn the nonlinear dynamics of the hydroturbine using neural controlled differential equations and then perturbs the obtained nonlinear system at different equilibrium points, based on which a static output feedback adaptive dynamic programming algorithm is then used to optimize the PID gains for each equilibrium point. Moreover, a continuous-time version of stochastic distribution control is proposed to further fine-tune the optimized PID gains. Finally, the controller is obtained by implementing linear interpolation between the optimized PID control gains. The simulation results show that the proposed gain-scheduling PID controller can control a larger range of operation points compared with the given fixed PID controller and the baseline method. Compared with the given fixed PID controller, the proposed gain-scheduling PID controller can regulate hydroturbine frequency against disturbances induced by power-load variation with over 50% less overshoot for some operation points.

13 HYDRO ENERGY

TRANSP integrated modeling code for interpretive and predictive analysis of tokamak plasmas

This paper provides a comprehensive review of the TRANSP code, a sophisticated tool for interpretive and predictive analysis of tokamak plasmas, detailing its major capabilities and features. It describes the equations for particle, power, and momentum balance analysis, as well as the poloidal field diffusion equations. The paper outlines the spatial and time grids used in TRANSP and details the equilibrium assumptions and solvers. Various models for heating and current drive and radiation, including updates to the NUBEAM model, are discussed. The handling of large-scale events such as sawtooth crashes and pellet injections is examined, along with the predictive capabilities for advancing plasma profiles. The integration of TRANSP with the ITER Integrated Modeling and Analysis Suite (IMAS) is highlighted, demonstrating enhanced data access and analysis capabilities. Additionally, the paper discusses best practices and continuous integration techniques to enhance TRANSP's robustness. The suite of TRANSP tools, designed for efficient data analysis and simulation, further supports the optimization of tokamak operations and coupling with other tokamak codes. Continuous development and support ensure that TRANSP remains a major code for the analysis of experimental data for controlled thermonuclear fusion, maintaining its critical role in supporting the optimization of tokamak operations and advancing fusion research.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY

Advancing continuous enzymatic hydrolysis for improved biomass saccharification

Background: A deployable, continuous enzymatic hydrolysis (CEH) process can address cost and commercialization risks associated with second-generation (Gen2) biorefinery sugar/lignin/ethanol production while contributing to energy supply and security. Developments in commercial enzymatic hydrolysis formulations targeting Gen2 pretreated biomass such as deacetylated mechanically refined (DMR) biomass necessitate a reassessment of the existing hybrid simultaneous saccharification and fermentation (SSF) approach. Notably, the practice of "finishing hydrolysis" in SSF has become problematic with the introduction of oxidative enzymes, such as lytic polysaccharide monooxygenases (LPMOs), into commercial cellulase formulations as these require specific redox conditions and cofactor. Moreover, continuous SSF has not been demonstrated at commercial scale, limiting deployment and the associated economic benefits to farmers, producers, and support industries. Results: Continuous enzymatic hydrolysis (CEH) was demonstrated at bench scale to enable optimal saccharification performance of deacetylated mechanically refined (DMR) pretreated biomass. Diafiltration was demonstrated to retain pretreated biomass solids and enzymes for continuous reaction while removing solubilized product sugars in situ. A significant breakthrough afforded by the CEH process is its ability to achieve equivalent endpoint conversions with approximately 50% lower enzyme loading. Yields of glucose and xylose were increased ~ 15% and ~ 4%, respectively, over batch hydrolysis. Unlike SSF using yeast or Zymomonas, CEH allows precise optimization of pH, temperature, oxygen tension, LPMO mediator concentration, and removal of end-product inhibitors. Conclusions: Advanced CEH holds promise as a transformational, process-intensified, and cost-effective method for producing soluble clarified biomass sugars and insoluble lignin-rich streams. Enhancing saccharification performance, optimizing operating parameters, and employing membrane filtration will help overcome existing challenges and enable the efficient production of valuable biomaterials from lignocellulosic biomass.

09 BIOMASS FUELS