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At least 163 records · Page 9

Microscopic dynamics of stress relaxation in a nanocolloidal soft glass

Here, we report x-ray photon correlation spectroscopy (XPCS) experiments with in situ rheometry performed on a soft glass composed of a concentrated suspension of charged silica nanoparticles subjected to step strains that induce yielding and flow. The XPCS measurements characterize the particle-scale and mesoscale motions within the glass that underlie the highly protracted decay of the macroscopic stress following the step strains. These dynamics are anisotropic, with slow, convective particle motion along the direction of the preceding shear that persists for surprisingly large times and that is accompanied by intermittent motion in the perpendicular (vorticity) direction. A close correspondence between the convective dynamics and stress relaxation is demonstrated by power-law scaling between the characteristic velocity of the collective particle motion and the rate of stress decay.

36 MATERIALS SCIENCE↗

Catalytic Reduction of Residual Oxygen for the Purification of Carbon Dioxide Streams from Pressurized Oxy-Combustors

Pressurized oxy-combustion (OC) is regarded as an energy-efficient alternative to atmospheric OC. To utilize the captured carbon dioxide (CO2) from pressurized OC for enhanced oil recovery or carbon storage in geological formations, the CO2 product stream must meet the required purity specifications. One major impurity in OC flue gases is residual O2 (e.g., 3 vol%). In this study, a catalytic approach has been investigated to remove the residual O2 through a reduction reaction with injected methane (CH4) in a reverse-flow fixed-bed (RFFB) reactor. The performance was tested for a supported copper (Cu)-based catalyst synthesized by incipient wetness impregnation and a cobalt-manganese (CoMn) oxide catalyst synthesized by a coprecipitation method. A commercial Cu-based catalyst was also studied as the reference. Experimental results revealed that under the reverse-flow operation, the temperature profiles in the reactor were symmetrical on both sides of the catalyst zone and were more homogenous than those under the conventional one-direction-flow operation. Among the tested catalysts, the CoMn catalyst was the most reactive for promoting the reduction of O2 with CH4, able to attain >95% O2 removal at 475 °C and a gas hourly space velocity (GHSV) of 18,000 h-1 with a feed gas containing 3 vol% O2. Under all experiments, the concentrations of carbon monoxide (CO) were lower than 30 ppmv in the effluent gas streams. The reaction between O2 and CH4 could be depicted by using the Mars–van Krevelen redox mechanism based on a two-step reduction–oxidation reaction pathway.

36 MATERIALS SCIENCE↗

Pulse Detonation Engine for Advanced Oxy-Combustion of Coal-Based Fuel for Direct Power Extraction Applications

Pressure gain combustion and magnetohydrodynamic (MHD) systems have the potential to provide a step increase in the efficiency of combined-cycle power plants. Specific advantages include a net pressure increase to the system instead of a pressure drop, the high temperature of the detonation waves can increase the efficiency of power extraction compared to other processes, significant thermal energy can be released in a compact region, and the high velocities of the flow increase extraction of electrical power. In summary, a pressure gain combustor coupled with a MHD has the potential to be transformative. Despite the potential advantages, relatively little research has been conducted considering coupled pressure gain combustion systems with MHD systems. With this background and motivation, the overall goal of this effort was to advance the knowledge, technology, and computational tools associated with coupled detonation and MHD systems. A joint experimental and computational approach was used while seeking to accomplish the goals of this work. Specifically, two pulse detonation engines were developed and used for the experiments to produce detonations. Detonation speeds were measured for a variety of flow and fuel conditions (e.g., methane, propane, with coal particles). Preliminary electrical conductivity measurements were collected. An extensive amount of research was performed to identify sensitivities of detonation behaviors to the presence of combustion products. Computationally, a twofold approach was used in this work. First, a solver was developed for solving the governing equations for a reactive flow with coupled detonation and ionization chemistry. The solver was applied to study the impacts of seed material ionization on detonation. Second, a conservation element-solution element (CE-SE) based numerical solver for detonation studies with a reduced reaction mechanism for oxy-methane combustion was developed and verified on standard test cases. Key findings and contributions from this work are as follows. A system was developed for injecting powderized coal, or other seeding material, into a pulse-detonation system. The influence of a combustion product (i.e., CO 2 ) on detonation behavior was identified. Knowledge gained from this work is applicable to devices such as rotating detonation engines, where combustion products mix with fresh reactants. A system for measuring the electrical conductivity of the exhaust from a pulse-detonation engine was developed. The open-source solver, Clawpack, was extended to solve the reactive Euler equations for simulating detonations. A coupled combustion and ionization chemistry was developed in a single chemical kinetic model for methane oxidation. This model can be used to solve coupled MHD and detonation simulations. It was found that parasitic interactions from ionization chemistry with the magnetic field can reduce the detonation velocity by up to 8%, with a potential impact on power extraction of 15%. It is recommended that interactions between the detonation front and MHD field be considered. Higher gas temperatures and velocities were achieved owing to oxy-fuel detonations. Use of radical dissociation reactions in the reduced reaction mechanism, was found to be critical in predicting detonation temperature and velocity accurately.

01 COAL, LIGNITE, AND PEAT↗

Update on ASME AG-1 and Revision of the DOE Nuclear Air Cleaning Handbook - 20536

The American Society of Mechanical Engineers AG-1 Code on Nuclear Air and Gas Treatment is updated every two years. This paper provides an overview of issues being discussed within AG-1 committee meetings and revisions that will be included in the next version of the code. Additional information is provided on issues being discussed ranging from media velocity limits, control of airflow using actual versus and standard cubic feet per minute. Navigation and identification of applicable high-efficiency particulate air (HEPA) filter code sections can be confusing for those unfamiliar with AG-1. A general reorganization of filtration code sections clarifies geometry, flow, and performance envelopes for nuclear grade HEPA filters. The initial step consists of establishment of Section FN covering qualification of conventional, ceramic, metal, and high strength fibrous glass media. Future filter sections are currently expected to be characterized by geometry and flow direction (axial vs. radial). The Department of Energy's (DOE) Nuclear Air Cleaning Handbook (NACH) is also undergoing revision. An update on the status of that effort, along with issues and changes being considered. The original charter of the AG-1 Code was specific to commercial nuclear power plants, and to safety-related air cleaning systems. This has evolved over the years to include normal non-safety ventilation systems and process gas systems. Corollary guidance for DOE facilities has been available in several editions of the Nuclear Air Cleaning Handbook, starting with ORNL-NSIC 65. The latest edition of the Handbook, 2003, is undergoing revision and modifications. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

A method for estimating light quenching in inorganic scintillator detectors for radioactive ion beam experiments

In recent experiments, inorganic scintillators have been used to study the decays of exotic nuclei, providing an alternative to silicon detectors and enabling measurements that were previously impossible. However, proper use of these materials requires us to understand and quantify the scintillation process, specifically in response to very heavy nuclei. Here, in this work, we show a simplified method based on the models of Birks (1951) and Meyer and Murray (1962) to parametrize the light output of inorganic scintillators in response to beams of energetic heavy ions over a broad range of energies. We test the accuracy of our parametrization approach by calculating light output and quenching factors for various ions and comparing them with experimental data from Lutetium Yttrium Orthosilicate (LYSO:Ce), a common inorganic scintillator. The Meyer–Murray model suggests that, for sufficiently heavy ions at high energies, the majority of the light output is associated with the creation of delta electrons, which are induced by the passage of the beam through the material. These delta electrons dramatically impact the response of detection systems when subject to ions with velocities typical of beams in modern fragmentation facilities. To illustrate this, we also present a qualitative estimate of the effects of delta rays on overall light output using the Birks–Meyer–Murray parametrization. The approach presented herein will serve as a basic framework for further, more rigorous studies of scintillator response to heavy ions. This work is a crucial first step in planning future experiments where energetic exotic nuclei are interacting with scintillator detectors.

Heavy ion↗

A spline-based method to obtain spatially dependent viscosity in confined flows

Coupling chemical physics to continuum theories is a critical step to understanding multi-scale phenomena. This paper will connect non-equilibrium molecular dynamics simulations to a continuum-based Navier-Stokes equation that has relaxed the assumption of spatial uniformity in viscosity. Using a form for viscosity based on spline interpolation, viscosity as a function of position is obtained from the least squares fit of the velocity profile measured from molecular simulations of flow in a nanochannel. Viscosity can vary widely, particularly near the channel boundaries, indicating that uniform viscosity is no longer appropriate. Variations of the viscosity near the channel surfaces imply that considering solution and surface chemistry could be necessary to rigorously understand molecular-scale flows in nanochannels.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Plasma Photonics: Manipulating Light Using Plasmas (Full Technical Final Report)

In this LDRD, we made significant advances on multiple fronts on the investigation and design of plasma-based optical structures to manipulate light at extreme intensities. We have unveiled new fundamental properties of laser-plasma interaction that were leveraged to introduce transformative applications. We first investigated new ways to generate high-amplitude plasma-based optical structures, based on the auto-resonance mechanism in order to produce refractive structures in plasmas. New insights into the interaction of such plasma structures with light waves allowed us to propose new methods for polarization manipulation and control for high-intensity lasers in the nonlinear regime, i.e. allowing the manipulation of an intense laser via its interaction with a less intense “control” laser beam in plasmas. Another application that we demonstrated experimentally is the capability to manipulate the velocity of light waves in plasmas by tailoring its refractive index; we reported the first demonstration of “slow light” and supra-luminal light propagation in plasmas. We also developed a new technological concept that would allow the probing of the type of plasma structures investigated in such applications, using high-bandwidth laser probing [Ludwig Phys. Plasmas 2019]. Finally, first steps were taken towards the demonstration of a plasma-based amplifier and compressor for the National Ignition Facility, in order to achieve powers equal of greater than those delivered by the Advanced Radiographic Capability (ARC). This technique uses a plasma-based grating acting like a compressor and amplifier, which could potentially compress and amplify an 88 ps, 56 J beam from the National Ignition Facility to <10 ps and hundreds of J.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Detailed biomass fast pyrolysis kinetics integrated to computational fluid dynamic (CFD) and discrete element modeling framework: Predicting product yields at the bench-scale

Fast pyrolysis is an intricate process due to the variability and anisotropy of lignocellulosic biomass and the complicated chemistry and physics during conversion in a bubbling fluidized bed reactor (BFBR). The complexity of biomass fast pyrolysis lends itself well to computational fluid dynamics (CFD) and discrete element (DEM) analysis, which promises to reduce experimental time and its associated cost. This work investigated switchgrass fast pyrolysis simulated by computational fluid dynamics coupled with a discrete element method to track individual reacting biomass particles throughout a bench-scale BFBR reactor. We accounted for the fast pyrolysis chemistry through a comprehensive reaction scheme with secondary cracking reactions. We performed a three-step reduction for secondary cracking reactions to convert the full cracking scheme into a reduced scheme easily incorporated into our model. We assessed the impact of operational conditions on the steady-state yields of liquid bio-oil, non-condensable gases (NCG), at 550 °C over a range of fluidization numbers (2 – 6 Umf), reported as a ratio to the minimum fluidization velocity (Umf). At steady-state, the volatile bio-oil yield had a range of 49.3–50.4 wt%. Levoglucosan was the primary volatile component present with 21 wt% of the bio-oil while water was the second largest with 20 wt%. The reduction of the secondary reaction schemes did not appreciably affect the overall yields of switchgrass pyrolysis compared to the full secondary scheme.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Numerical simulation of vortex-induced vibration response of a single IEA 10-MW wind turbine blade

Three-dimensional simulation of vortex-induced vibration (VIV) of a single International Energy Agency (IEA) 10-MW reference wind turbine blade with a length of 97.325 m is performed using the ExaWind stack, an open-source suite of codes. This study aims to illustrate the spanwise VIV response characteristics and cross-validate the results with an existing commercial framework. Five near-body meshes and three time steps are selected for the convergence study. To improve computational efficiency, several VIV triggering methods are also compared to shorten the VIV development period. The ExaWind-based VIV simulation strategy for a single IEA 10-MW blade is determined. First, the modal shape is validated against published results. Then, spanwise VIV responses of four blade configurations under a fixed and varied incoming flow velocity are analyzed. Results show that the VIV response is dominated by the first edgewise (second overall) mode. Little first-mode contributions appear near the second-mode node, producing a pi phase jump, and a higher harmonics response occurs near the blade root. Rotational degrees of freedom are minor compared with translational motion. The response versus reduced velocity is analyzed, showing a two-branch behavior similar to that of VIV for a bluff cylinder. Across all tested cases, the dominant frequency remains locked to the natural frequency of the second mode with no observed desynchronization. A mild deviation is observed for the case of 90-degree pitch and 310-degree azimuth rotation near a reduced velocity of 6, which will be examined with additional cases in future work. These findings indicate that severe VIV responses can arise under specific configurations and flow conditions, thereby increasing the potential for VIV fatigue damage and requiring greater attention during operation.

17 WIND ENERGY↗

Vortical interactions in turbulent thermoacoustic systems

This study examines the dynamics of vortical interactions and their implications for mitigating thermoacoustic instability in a turbulent combustor. The regions of intense vortical interactions are identified as vortical communities in the network space of weighted directed vortical networks constructed from two-dimensional experimental velocity data. One can expect vortical interactions in the combustor to be strongest near the moment of vortex shedding, as the shed vortices gradually weaken due to dissipation while convecting downstream. However, we show that, during the state of thermoacoustic instability, there is a non-trivial consistent phase lag of approximately 52° between the shedding of the coherent structures from the backward-facing step and the time instant when the vortical interactions attain their local maximum value. We explain this phase lag by investigating the correlation between acoustic pressure fluctuations, spatio-temporal dynamics of coherent structures and vortical interactions in the reaction field of the combustor. We also show the aperiodic variation of vortical interactions during the states of combustion noise and aperiodic epochs of intermittency. Furthermore, the spatio-temporal evolution of pairs of vortical communities with the maximum inter-community interactions provides insight into explaining the critical regions detected in the reaction field during the states of intermittency and thermoacoustic instability, also identified in previous studies. As a result, we further show that the most efficient suppression of thermoacoustic instability via air microjet injection is achieved when steady air jets are introduced to disrupt the maximum inter-community interactions present during the state of thermoacoustic instability.

Sahay, Ankit [Indian Institute of Technology Madra↗

A TOPAS model for lens-based proton radiography

Abstract Objective. Proton Radiography can be used in conjunction with proton therapy for patient positioning, real-time estimates of stopping power, and adaptive therapy in regions with motion. The modeling capability shown here can be used to evaluate lens-based radiography as an instantaneous proton-based radiographic technique. The utilization of user-friendly Monte Carlo program TOPAS enables collaborators and other users to easily conduct medical- and therapy- based simulations of the Los Alamos Neutron Science Center (LANSCE). The resulting transport model is an open-source Monte Carlo package for simulations of proton and heavy ion therapy treatments and concurrent particle imaging. Approach. The four-quadrupole, magnetic lens system of the 800-MeV proton beamline at LANSCE is modeled in TOPAS. Several imaging and contrast objects were modelled to assess transmission at energies from 230–930 MeV and different levels of particle collimation. At different proton energies, the strength of the magnetic field was scaled according to βγ, the inverse product of particle relativistic velocity and particle momentum. Main results. Materials with high atomic number, Z, (gold, gallium, bone-equivalent) generated more contrast than materials with low-Z (water, lung-equivalent, adipose-equivalent). A 5-mrad collimator was beneficial for tissue-to-contrast agent contrast, while a 10-mrad collimator was best to distinguish between different high-Z materials. Assessment with a step-wedge phantom showed water-equivalent path length did not scale directly according to predicted values but could be mapped more accurately with calibration. Poor image quality was observed at low energies (230 MeV), but improved as proton energy increased, with sub-mm resolution at 630 MeV. Significance. Proton radiography becomes viable for shallow bone structures at 330 MeV, and for deeper structures at 630 MeV. Visibility improves with use of high-Z contrast agents. This modality may be particularly viable at carbon therapy centers with accelerators capable of delivering high energy protons and could be performed with carbon therapy.

60 APPLIED LIFE SCIENCES↗

Resolution requirements for energy conservation in kinetic plasma simulations

The kinetic codes used to model the coupled dynamics of electromagnetic fields and charged particle transport have requirements for spatial, temporal, and charge resolution. These requirements may vary by the solution technique and scope of the problem. In this report, we investigate the resolution limits in the energy-conserving implicit particle-in-cell code CHICAGO. This report has the narrow aim of determining the maximum acceptable grid spacing for the dense plasmas generated in models of z-pinch target gases and power-flow electrode plasmas. In the 2D sample problem, the plasma drifts without external forces with velocity of 10 cm/µs. Simulations are scaled by plasma density to maintain uniform strides across the plasma and from the plasma to the boundaries. Additionally, the cloud-in-cell technique is used with 400 particles per cell and Δ t = 0.85× the Courant limit. For the linear cloud distribution, the criterion for conserving energy is Δ E/E tot < 0.01 for 50,000 time steps. The grid resolution is determined to crudely be Δx ≲ 3 l s , where l s is the electron collisionless skin depth. For the second-order cloud distribution the criterion is Δ E/E tot < 0.005 yielding Δx ≤ 15 l s . These scalings are functions of the chosen v d , Δ t , particles-per-cell, and number of steps.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Where Is the Provenance? Ethical Replicability and Reproducibility in GIScience and Its Critical Applications

As replicability and reproducibility (R&R) crises develop within emerging convergent inquiry, ethical use of provenance information is central to the establishment and preservation of trust in critical applications of GIScience and geospatial technologies. Today large volumes of geospatial data are generated at high velocity from satellite sensors and unmanned aircraft systems, citizen sensors, geolocation-based data services, global navigation satellite systems, and so on. The extensive use of these data for applications such as disaster and humanitarian response raises the issue of R&R from competing perspectives of location privacy and geospatial data quality. Although geospatial data can be integrated and linked with contextual information to identify individuals’ movements, steps taken to ensure privacy can complicate the multiuser development of high-quality geospatial workflows. Provenance information as digital records of historical (retrospective) and potential future (prospective) geospatial processes is often overlooked, misunderstood, or inadequately addressed. We explore the relationship between provenance information, location privacy, and geospatial data quality in the context of R&R with a focus on disaster analytics. Here, we argue that in the era of big data and deep learning, GIScientists and associated institutions bear greater responsibility both for geospatial workflow quality and for location privacy. Given vastly heterogenous computational landscapes, we provide practical recommendations for ethically driven provenance and R&R research and development within the GIScience community and beyond.

96 KNOWLEDGE MANAGEMENT AND PRESERVATION↗

Efficient Loads Surrogates for Waked Turbines in an Array

Accurately and efficiently predicting wind turbine structural loading is a crucial step in wind farm design. Without considering structural loading, wind farm optimization could negatively impact turbine fatigue and ultimate loads, especially for waked and partially waked turbines, which could result in higher maintenance costs and reduced turbine lifetime. However, predicting turbine loads throughout an array is a costly step, as these quantities require time-accurate results across long time histories, which is often intractable for large array optimization. Therefore, surrogate models that link array spacing to load outputs are often used, but the surrogates are then unique to the inflow conditions and array configurations in the training library. This work develops surrogate models for many wind turbine load outputs based solely on rotor plane velocity measurements, with no required input about array configuration or freestream inflow parameters. Surrogate models were constructed for many turbine quantities of interest (QoI), considering mean, standard deviation, ultimate, and fatigue loads. In general, most QoI statistics were accurately captured, as measured by predicted vs. actual correlation coefficient, confirming the suitability of the approach. Temporal mean values of the QoI required only temporal mean measurements of the rotor plane inflow velocity. However, accurate prediction of temporal standard deviation, ultimate, and fatigue values of QoI also required temporal standard deviations of the rotor plane velocity field. Poor surrogate performance was observed when too many correlated inputs were used, such as multiple velocity components. If the fewest inflow parameters are used to construct the surrogates, the average correlation coefficient value for all output QoI statistics is 0.89. Surrogates for standard deviations and damage equivalent loads (DELs) of turbine QoIs generally had lower accuracy and tower-base and shaft load channels posed the most difficult to capture accurately. The results suggest that these surrogates could be easily paired with analytic wake models, which are frequently used for pre-construction wind farm array optimization, to account for turbine loading in addition to power production. By including the optimal inflow conditions, the surrogate accuracy can improve to an average correlation coefficient value for all output QoI statistics of 0.92. This work has established the ability to build accurate surrogates for mean, standard deviation, ultimate load, and DEL turbine QoI values based on the rotor plane inflow velocity, and identified which inflow conditions lead to greater surrogate accuracy.

17 WIND ENERGY↗

Semi-implicit continuum kinetic modeling of weakly collisional parallel transport in a magnetic mirror

We present implicit-explicit (IMEX) kinetic simulations of weakly collisional parallel plasma transport in magnetic mirror configurations using the continuum code COGENT. The numerical scheme employs a Jacobian-free Newton–Krylov method with algebraic multigrid preconditioning to overcome the severe time step limitations imposed by strong mirror forces in fully explicit schemes. Applied to parameters relevant to the Wisconsin HTS Axisymmetric Mirror experiment, the IMEX approach enables time steps up to 2.5×10 4 times larger than those permitted by explicit methods, resulting in a 2500× speedup in 1D–2V simulations of parallel transport with kinetic ions and Boltzmann electrons. Additionally, a reduced bounce-averaged model for a square mirror is implemented to support the computationally intensive fully kinetic simulations. The bounce-averaged formulation is used to evaluate the numerical convergence of the velocity-space discretization algorithms and to assess the role of the collision model by comparing simulations employing the nonlinear Fokker–Planck and the simplified Lenard–Bernstein–Dougherty collision operators.

Collision theories↗

Velocity Extraction Using Complete Time-Domain Waveform Data and Audio Machine Learning

We developed a new machine learning-based tool for extracting information from interferometry measurements: MIDWAZE (Modular Interferometry Direct Waveform AnalyZEr). This paper showcases MIDWAZE’s ability to extract an object’s velocity information from Photonic Doppler Velocimetry (PDV) data at near-human accuracy with little to no human intervention. MIDWAZE can extract velocities roughly 350 times as fast as a human analyst "rushing" to complete their extractions, with similar extraction accuracy. MIDWAZE’s most outstanding feature is that it operates directly in waveform/temporal space, freeing analysis from certain limitations imposed by traditional spectrogram-based approaches and opening the way to "phase aware" PDV analysis. MIDWAZE also has limited ability to discriminate between different solid objects, which we develop as a first step towards automated discrimination of different kinds of objects such as ejecta clouds.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Initial Characterization of the NREL Large-Amplitude Motion Platform

The Large Amplitude Motion Platform (LAMP) at NREL represents a significant advancement in the controlled testing of Wave Energy Converters (WECs) under laboratory conditions. Originally designed by E2M as a six-degree-of-freedom (DOF) Stewart platform for flight simulation, LAMP has been adapted by NREL to facilitate the mounting and evaluation of WECs. This adaptation enables dry testing of WECs using motion profiles similar to the ocean, facilitating the iterative design, testing, and validation of WEC performance prior to ocean deployments. This report presents the initial work completed to characterize LAMP, with particular emphasis on its stability and operational capabilities across various single and multi-degree-of-freedom (DOF) motion profiles. The report includes planned comparisons at three distinct mass payloads, aimed at assessing the platform's positional accuracy, frequency response, and endurance over extended runtime periods. These experimental tests are critical for establishing the platform's limitations and ensuring that the data generated during WEC validation is both accurate and reproducible. The outcomes of this study not only contribute to a deeper understanding of LAMP's capabilities but also lay the groundwork for future advancements in WEC testing methodologies. By providing robust and reliable performance data within a controlled laboratory setting, the findings are expected to significantly enhance the development and commercialization of marine energy technologies. This report presents the initial findings of the LAMP Characterization work and proposed steps to further understand and characterize LAMP. Data collected during this work can be found on MHKDR at: https://mhkdr.openei.org/submissions/602 Note that this report shares the measured/found instantaneous maximum operating range of LAMP. For most applications the maximum operating range cannot be used for system health and longevity. The operating range and capabilities of LAMP will be evaluated on a case-by-case basis, single-DOF position, velocity, and acceleration values presented in Table 4a-c, and Table 5 should be taken as instantaneous absolute maximum values. Future use of LAMP will likely be limited to smaller values.

16 TIDAL AND WAVE POWER↗

Single-molecule tracking reveals dual front door/back door inhibition of Cel7A cellulase by its product cellobiose

Degrading cellulose is a key step in the processing of lignocellulosic biomass into bioethanol. Cellobiose, the disaccharide product of cellulose degradation, has been shown to inhibit cellulase activity, but the mechanisms underlying product inhibition are not clear. We combined single-molecule imaging and biochemical investigations with the goal of revealing the mechanism by which cellobiose inhibits the activity of Trichoderma reesei Cel7A, a well-characterized exo-cellulase. We find that cellobiose slows the processive velocity of Cel7A and shortens the distance moved per encounter; effects that can be explained by cellobiose binding to the product release site of the enzyme. Cellobiose also strongly inhibits the binding of Cel7A to immobilized cellulose, with a K i of 2.1 mM. The isolated catalytic domain (CD) of Cel7A was also inhibited to a similar degree by cellobiose, and binding of an isolated carbohydrate-binding module to cellulose was not inhibited by cellobiose, suggesting that cellobiose acts on the CD alone. Finally, cellopentaose inhibited Cel7A binding at micromolar concentrations without affecting the enzyme’s velocity of movement along cellulose. Together, these results suggest that cellobiose inhibits Cel7A activity both by binding to the “back door” product release site to slow activity and to the “front door” substrate-binding tunnel to inhibit interaction with cellulose. These findings point to strategies for engineering cellulases to reduce product inhibition and enhance cellulose degradation, supporting the growth of a sustainable bioeconomy.

59 BASIC BIOLOGICAL SCIENCES↗