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NASA NTRS2010

Experimental Space Shuttle Orbiter Studies to Acquire Data for Code and Flight Heating Model Validation

In an experimental study to obtain detailed heating data over the Space Shuttle Orbiter, CUBRC has completed an extensive matrix of experiments using three distinct models and two unique hypervelocity wind tunnel facilities. This detailed data will be employed to assess heating augmentation due to boundary layer transition on the Orbiter wing leading edge and wind side acreage with comparisons to computational methods and flight data obtained during the Orbiter Entry Boundary Layer Flight Experiment and HYTHIRM during STS-119 reentry. These comparisons will facilitate critical updates to be made to the engineering tools employed to make assessments about natural and tripped boundary layer transition during Orbiter reentry. To achieve the goals of this study data was obtained over a range of Mach numbers from 10 to 18, with flight scaled Reynolds numbers and model attitudes representing key points on the Orbiter reentry trajectory. The first of these studies were performed as an integral part of Return to Flight activities following the accident that occurred during the reentry of the Space Shuttle Columbia (STS-107) in February of 2003. This accident was caused by debris, which originated from the foam covering the external tank bipod fitting ramps, striking and damaging critical wing leading edge heating tiles that reside in the Orbiter bow shock/wing interaction region. During investigation of the accident aeroheating team members discovered that only a limited amount of experimental wing leading edge data existed in this critical peak heating area and a need arose to acquire a detailed dataset of heating in this region. This new dataset was acquired in three phases consisting of a risk mitigation phase employing a 1.8% scale Orbiter model with special temperature sensitive paint covering the wing leading edge, a 0.9% scale Orbiter model with high resolution thin-film instrumentation in the span direction, and the primary 1.8% scale Orbiter model with detailed thin-film resolution in both the span and chord direction in the area of peak heating. Additional objectives of this first study included: obtaining natural or tripped turbulent wing leading edge heating levels, assessing the effectiveness of protuberances and cavities placed at specified locations on the orbiter over a range of Mach numbers and Reynolds numbers to evaluate and compare to existing engineering and computational tools, obtaining cavity floor heating to aid in the verification of cavity heating correlations, acquiring control surface deflection heating data on both the main body flap and elevons, and obtain high speed schlieren videos of the interaction of the orbiter nose bow shock with the wing leading edge. To support these objectives, the stainless steel 1.8% scale orbiter model in addition to the sensors on the wing leading edge was instrumented down the windward centerline, over the wing acreage on the port side, and painted with temperature sensitive paint on the starboard side wing acreage. In all, the stainless steel 1.8% scale Orbiter model was instrumented with over three-hundred highly sensitive thin-film heating sensors, two-hundred of which were located in the wing leading edge shock interaction region. Further experimental studies will also be performed following the successful acquisition of flight data during the Orbiter Entry Boundary Layer Flight Experiment and HYTHIRM on STS-119 at specific data points simulating flight conditions and geometries. Additional instrumentation and a protuberance matching the layout present during the STS-119 boundary layer transition flight experiment were added with testing performed at Mach number and Reynolds number conditions simulating conditions experienced in flight. In addition to the experimental studies, CUBRC also performed a large amount of CFD analysis to confirm and validate not only the tunnel freestream conditions, but also 3D flows over the orbiter acreage, wing leading edge, and controlurfaces to assess data quality, shock interaction locations, and control surface separation regions. This analysis is a standard part of any experimental program at CUBRC, and this information was of key importance for post-test data quality analysis and understanding particular phenomena seen in the data. All work during this effort was sponsored and paid for by the NASA Space Shuttle Program Office at the Johnson Space Center in Houston, Texas.

Wadhams, T. P.↗

DOE OSTI2020

Limitations of traditional tools for beyond design basis external hazard PRA

Probabilistic risk assessment (PRA) is being used increasingly by the nuclear industry for safety during normal operations as well as for the protection against external hazards. Computation of total risk in an external hazard PRA is dependent on hazard assessment, fragility assessment, and systems analysis. A systems analysis for propagation of component fragilities is conducted using event and fault trees. The event and fault trees for an actual power plant can be fairly large in size, which imposes computational challenges. Hence, certain assumptions are employed for computational efficiency. These assumptions typically represent the conditions imposed during the design basis (DB) scenario. The traditional PRA tools based on these assumptions are also widely applied to perform risk assessment in the context of beyond design basis (BDB) scenarios. However, some of these assumptions may not be valid for certain BDB scenarios. In addition, the probability of dependent failures also increases in BDB scenarios due to common cause failures (CCF) which usually results from design modifications, human errors, etc. In this manuscript, a simple and a relatively more complex illustrative examples are used to show the limitation of these assumptions in the numerical quantification of risk for the case of BDB conditions. Case studies with CCF events across multiple fault trees are also presented to illustrate the effect of these assumptions when traditional approach is used in BDB risk assessment. It is shown that the assumptions are valid for the case of DB conditions but may lead to excessively conservative risk estimates in the case of BDB conditions. Finally, a Bayesian network based top-down algorithm is proposed as an alternative tool for accurate numerical quantification of total risk in systems analysis.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗

DOE OSTI2019

Preliminary Design Study of a Fast-Ramping Magnet for Pre-Concept Design of an Electron-Ion Collider at Jefferson Lab

The Jefferson Lab Electron Ion Collider (JLEIC) is a proposed new machine for nuclear physics research. The all new ion accelerator and collider complex will consist of two collider rings with a unique figure-of-eight layout to deliver a high degree of polarization in both beams. As part of the pre-concept design for the Ion ring, a 3-Tesla Super-Ferric dipole magnet was proposed utilizing a superconducting Cable-in-Conduit-Conductor (CICC) design to wind the coils which will be built by Texas A&M University. A first mechanical model of the winding for the 3T-SF-CICC dipole was built to validate that the winding structure provides the conductor geometry required to provide collider field homogeneity over large aperture. A rapid-cycling Booster synchrotron is required to inject 8 GeV beams to the Ion Ring. The Booster requires arc dipoles with the same field and aperture as those of the Ion Ring. Due to the design of the CICC with respect to the amount of stabilizer and the internal cooling mechanism employed, temperature rise of the coils during a quench event is much more rapid than for more conventional magnets. It is thus imperative that the magnet?s stored energy is dissipated externally to the windings to avoid overheating the CICC and to provide adequate protection during a quench. As a result, this paper presents a preliminary design study, including AC effects inside the coils and associated risks with the aim of providing guidance for the design of the full protection system for such a magnet.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

NASA NTRS2013

Investigation of a Cross-Correlation Based Optical Strain Measurement Technique for Detecting radial Growth on a Rotating Disk

The Aeronautical Sciences Project under NASA`s Fundamental Aeronautics Program is extremely interested in the development of novel measurement technologies, such as optical surface measurements in the internal parts of a flow path, for in situ health monitoring of gas turbine engines. In situ health monitoring has the potential to detect flaws, i.e. cracks in key components, such as engine turbine disks, before the flaws lead to catastrophic failure. In the present study, a cross-correlation imaging technique is investigated in a proof-of-concept study as a possible optical technique to measure the radial growth and strain field on an already cracked sub-scale turbine engine disk under loaded conditions in the NASA Glenn Research Center`s High Precision Rotordynamics Laboratory. The optical strain measurement technique under investigation offers potential fault detection using an applied high-contrast random speckle pattern and imaging the pattern under unloaded and loaded conditions with a CCD camera. Spinning the cracked disk at high speeds induces an external load, resulting in a radial growth of the disk of approximately 50.0-im in the flawed region and hence, a localized strain field. When imaging the cracked disk under static conditions, the disk will be undistorted; however, during rotation the cracked region will grow radially, thus causing the applied particle pattern to be .shifted`. The resulting particle displacements between the two images will then be measured using the two-dimensional cross-correlation algorithms implemented in standard Particle Image Velocimetry (PIV) software to track the disk growth, which facilitates calculation of the localized strain field. In order to develop and validate this optical strain measurement technique an initial proof-of-concept experiment is carried out in a controlled environment. Using PIV optimization principles and guidelines, three potential speckle patterns, for future use on the rotating disk, are developed and investigated in the controlled experiment. A range of known shifts are induced on the patterns; reference and data images are acquired before and after the induced shift, respectively, and the images are processed using the cross-correlation algorithms in order to determine the particle displacements. The effectiveness of each pattern at resolving the known shift is evaluated and discussed in order to choose the most suitable pattern to be implemented onto a rotating disk in the Rotordynamics Lab. Although testing on the rotating disk has not yet been performed, the driving principles behind the development of the present optical technique are based upon critical aspects of the future experiment, such as the amount of expected radial growth, disk analysis, and experimental design and are therefore addressed in the paper.

Clem, Michelle M.↗

NASA NTRS1988

Evaluation of flammability for cable-like polymers

A new test method is developed for the flammability of insulated electric cables as well as polymers formed in the cables. The rate of downward flame spread along the polymer surface is measured by a technique in which the flame is stopped at a particular position by winding up the cable at the same speed as the flame spreading rate, with external preheating by an electric furnace having oxidative gas flow. The polymer burns continuously without change of flame condition and the rate is obtained from the winding speed of the cable. A flame spread equation relates the flame spreading rate (V) of the cable-like polymer to O concentration (Yo) and the atmosphere, the preheating temperature, and the pyrolysis temperature for polymer burning. Plotting log V vs. log Yo for experimental results allows evaluation of the flammability of polymers, with discrimination between the effects of O concentration, heating temperature, and specimen shape. Results with various types of PVC electrical cable indicated the validity of the method. Application of the method to the cables in groups showed a flame retardation behavior different from the case of a single cable, in a atmosphere of high O concentration.

Mikado, Tuneo↗

NASA NTRS2005

Pressurization System Modeling for a Generic Bimese Two- Stage-to-Orbit Reusable Launch Vehicle

A pressurization system model was developed for a generic bimese Two-Stage-to-orbit Reusable Launch Vehicle using a cross-feed system and operating with densified propellants. The model was based on the pressurization system model for a crossfeed subscale water test article and was validated with test data obtained from the test article. The model consists of the liquid oxygen and liquid hydrogen pressurization models, each made up of two submodels, Booster and Orbiter tank pressurization models. The tanks are controlled within a 0.2-psi band and pressurized on the ground with ambient helium and autogenously in flight with gaseous oxygen and gaseous hydrogen. A 15-psi pressure difference is maintained between the Booster and Orbiter tanks to ensure crossfeed check valve closure before Booster separation. The analysis uses an ascent trajectory generated for a generic bimese vehicle and a tank configuration based on the Space Shuttle External Tank. It determines the flow rates required to pressurize the tanks on the ground and in flight, and demonstrates the model's capability to analyze the pressurization system performance of a full-scale bimese vehicle with densified propellants.

Mazurkivich, Pete↗

NASA NTRS2008

Deflection Analysis of the Space Shuttle External Tank Door Drive Mechanism

Upon observing an abnormal closure of the Space Shuttle s External Tank Doors (ETD), a dynamic model was created in MSC/ADAMS to conduct deflection analyses of the Door Drive Mechanism (DDM). For a similar analysis, the traditional approach would be to construct a full finite element model of the mechanism. The purpose of this paper is to describe an alternative approach that models the flexibility of the DDM using a lumped parameter approximation to capture the compliance of individual parts within the drive linkage. This approach allows for rapid construction of a dynamic model in a time-critical setting, while still retaining the appropriate equivalent stiffness of each linkage component. As a validation of these equivalent stiffnesses, finite element analysis (FEA) was used to iteratively update the model towards convergence. Following this analysis, deflections recovered from the dynamic model can be used to calculate stress and classify each component s deformation as either elastic or plastic. Based on the modeling assumptions used in this analysis and the maximum input forcing condition, two components in the DDM show a factor of safety less than or equal to 0.5. However, to accurately evaluate the induced stresses, additional mechanism rigging information would be necessary to characterize the input forcing conditions. This information would also allow for the classification of stresses as either elastic or plastic.

Tosto, Michael A.↗

NASA NTRSDate not supplied

Flow Fields of Internally Mixed Exhaust Systems With External Plug For Supersonic Transport Applications

Commercial supersonic vehicles of the future will likely use engines with lower bypass ratios, where the design of the internal mixer will have a strong impact on the noise produced by the jet plume. Their exhaust systems may also feature external plugs to improve boattail angle for cruise performance at supersonic speeds. Currently there are no publicly available empirical noise models for such nozzle systems, and insight into their flow fields will help in creating these models. For those attempting to make large eddy simulations and other higher fidelity methods be their main prediction tool, the internally mixed exhaust system is also a good test case when going beyond simple single-stream jets. The turbulent flow statistics of several configurations previously tested for noise and shocks have been measured for flow conditions that can be used in development of empirical models and validating scale-resolving prediction tools. These measurements are presented and briefly analyzed for insights into the noise impacts from the flow impacts observed.

Turbulence, Jets, Particle Image velocimetry↗

NASA NTRSDate not supplied

Flow Fields of Internally Mixed Exhaust Systems With External Plug For Supersonic Transport Applications

Fluid Mechanics and Thermodynamics↗

NASA NTRS1995

The JPL Field Emission Spectrometer

The Jet Propulsion Laboratory (JPL) Field Emission Spectrometer (FES) was built by Designs and Prototypes based on a set of functional requirements supplied by JPL. The instrument has a spectral resolution of 6 wavenumbers (wn) and can acquire spectra from either the Mid Infrared (3-5 mu m) or the Thermal Infrared (8-12 pm) depending on whether the InSb or HgCdTe detector is installed respectively. The instrument consists of an optical head system unit and battery. The optical head which is tripod mounted includes the interferometer and detector dewar assembly. Wavelength calibration of the interferometer is achieved using a Helium-Neon laser diode. The dewar needs replenishing with liquid Nitrogen approximately every four hours. The system unit includes the controls for operation and the computer used for acquiring viewing and processing spectra. Radiometric calibration is achieved with an external temperature-controlled blackbody that mounts on the fore-optics of the instrument. The blackbody can be set at 5 C increments between 10 and 55 C. The instrument is compact and weighs about 33 kg. Both the wavelength calibration and radiometric calibration of the instrument have been evaluated. The wavelength calibration was checked by comparison of the position of water features in a spectrum of the sky with their position in the output from a high resolution atmospheric model. The results indicatethat the features in the sky spectrum are within 6-8 wn of their position ill the model spectrum. The radiometric calibration was checked by first calibrating the instrument using the external blackbody supplied with the instrument and then measuring the radiance from another external blackbody at a series of temperatures. The temperatures of these radiance spectra were then recovered by inventing Planck's law and the recovered temperatures compared lo the measured blackbody temperature. These results indicate that radiometric calibration is good to 0.5 C over the range of temperatures 10 to 55 C. The results also indicate that the instrument drifts slowly over time and should be recalibrated every 20 to 30 minutes in the field to ensure good radiometric fidelity. The instrument has now been extensively tested in the field in the United States and Australia. These in situ field measurements are being used to validate emissivity spectra recovered from the Thermal Infrared Multispectral Scanner (TIMS) and also the Australian CO2 Laser. The availability of in situ measurements is proving crucial to validation of the spectra derived from the airborne instruments since many natural surfaces cannot be easily transported back to the laboratory.

Hook, Simon J.↗

DOE OSTI2020

Residually Stressed Bimaterial Beam Specimen for Measuring Environmentally Assisted Crack Growth

Background: Subcritical crack growth can occur in a brittle material when the stress intensity factor is smaller than the fracture toughness if an oxidizing agent (such as water) is present at the crack tip. Objective: Here we present a novel bi-material beam specimen which can measure environmentally assisted crack growth rates. The specimen is “self-loaded” by residual stress and requires no external loading. Methods: Two materials with different coefficient of thermal expansion are diffusion bonded at high temperature. After cooling to room temperature a subcritical crack is driven by thermal residual stresses. A finite element model is used to design the specimen geometry in terms of material properties in order to achieve the desired crack tip driving force. Results: The specimen is designed so that the crack driving force decreases as the crack extends, thus enabling the measurement of the crack velocity versus driving force relationship with a single test. The method is demonstrated by measuring slow crack growth data in soda lime silicate glass and validated by comparison to previously published data. Conclusions: The self-loaded nature of the specimen makes it ideal for measuring the very low crack velocities needed to predict brittle failure at long lifetimes.

36 MATERIALS SCIENCE↗

DOE OSTI2018

Reconstruction of the complex angle-dependent photoionization transition dipole from a laser-dressed streaking experiment

We study the retrieval of the angular dependence of the photoionization transition dipole phase of an attosecond XUV pulse in order to reconstruct the whole photoelectron wave packet after the photoemission process. Using a one-electron model Ar atom, we show that the full electron wave packet can be reconstructed from IR-free photoelectron angular distributions and the streaking electron spectra in an IR-dressed XUV photoionization, with photoelectrons along the common polarization direction of the two pulses. The method relies on the validity of the strong-field approximation so it is valid only for higher energy photoelectrons. Here, we also point out that a single photoionization group time delay is not enough to represent a photoemission process; instead, a complete characterization of the electron wave packet requires the retrieval of the spectral phase over the whole bandwidth of the wave packet.

74 ATOMIC AND MOLECULAR PHYSICS↗

DOE OSTI2021

Deep learning of accurate force field of ferroelectric HfO 2

The discovery of ferroelectricity in HfO 2 -based thin films opens up new opportunities for using this silicon-compatible ferroelectric to realize low-power logic circuits and high-density nonvolatile memories. The functional performances of ferroelectrics are intimately related to their dynamic responses to external stimuli such as electric fields at finite temperatures. Molecular dynamics is an ideal technique for investigating dynamical processes on large length and time scales, though its applications to new materials are often hindered by the limited availability and accuracy of classical force fields. Here we present a deep neural network–based interatomic force field of HfO 2 learned from ab initio data using a concurrent learning procedure. The model potential is able to predict structural properties such as elastic constants, equation of states, phonon dispersion relationships, and phase transition barriers of various hafnia polymorphs with accuracy comparable with density functional theory calculations. Here, the validity of this model potential is further confirmed by the reproduction of experimental sequences of temperature-driven ferroelectric-paraelectric phase transitions of HfO 2 with isobaric-isothermal ensemble molecular dynamics simulations. We suggest a general approach to extend the model potential of HfO 2 to related material systems including dopants and defects.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

NASA NTRS2004

TiAl Scramjet Inlet Flap Subelement Designed and Fabricated

Next-generation launch vehicles are being designed with turbine-based combined cycle (TBCC) propulsion systems having very aggressive thrust/weight targets and long lives. Achievement of these goals requires advanced materials in a wide spectrum of components. TiAl has been identified as a potential backstructure material for maintainable composite panel heat exchangers (HEX) in the inlet, combustor, and nozzle section of a TBCC propulsion system. Weight reduction is the primary objective of this technology. Design tradeoff studies have assessed that a TiAl structure, utilizing a high-strength, hightemperature TiAl alloy called Gamma MET PX,1 reduce weight by 41 to 48 percent in comparison to the baseline Inconel 718 configuration for the TBCC propulsion system inlet, combustor, and nozzle. A collaborative effort between the NASA Glenn Research Center, Pratt & Whitney, Engineering Evaluation & Design, PLANSEE AG (Austria), and the Austrian Space Agency was undertaken to design, manufacture, and validate a Gamma-MET PX TiAl structure for scramjet applications. The TiAl inlet flap was designed with segmented flaps to improve manufacturability, to better control thermal distortion and thermal stresses, and to allow for maintainable HEX segments. The design philosophy was to avoid excessively complicated shapes, to minimize the number of stress concentrations, to keep the part sizes reasonable to match processing capabilities, and to avoid risky processes such as welding. The conceptual design used a standard HEX approach with a double-pass coolant concept for centrally located manifolds. The flowpath side was actively cooled, and an insulation package was placed on the external side to save weight. The inlet flap was analyzed structurally, and local high-stress regions were addressed with local reinforcements.

Draper, Susan L.↗

NASA NTRS2017

The Integrated Medical Model: Outcomes from Independent Review

In 2016, the Integrated Medical Model (IMM) v4.0 underwent an extensive external review in preparation for transition to an operational status. In order to insure impartiality of the review process, the Exploration Medical Capabilities Element of NASA's Human Research Program convened the review through the Systems Review Office at NASA Goddard Space Flight Center (GSFC). The review board convened by GSFC consisted of persons from both NASA and academia with expertise in the fields of statistics, epidemiology, modeling, software development, aerospace medicine, and project management (see Figure 1). The board reviewed software and code standards, as well as evidence pedigree associated with both the input and outcomes information. The board also assesses the models verification, validation, sensitivity to parameters and ability to answer operational questions. This talk will discuss the processes for designing the review, how the review progressed and the findings from the board, as well as summarize the IMM project responses to those findings. Overall, the board found that the IMM is scientifically sound, represents a necessary, comprehensive approach to identifying medical and environmental risks facing astronauts in long duration missions and is an excellent tool for communication between engineers and physicians. The board also found IMM and its customer(s) should convene an additional review of the IMM data sources and to develop a sustainable approach to augment, peer review, and maintain the information utilized in the IMM. The board found this is critically important because medical knowledge continues to evolve. Delivery of IMM v4.0 to the Crew Health and Safety (CHS) Program will occur in the 2017. Once delivered for operational decision support, IMM v4.0 will provide CHS with additional quantitative capability in to assess astronaut medical risks and required medical capabilities to help drive down overall mission risks.

medical equipment↗

DOE OSTI2025

Operational optimization for multi-functional charging station with electric and hydrogen-powered vehicles

The rapid adoption of electric vehicles (EVs) and hydrogen fuel cell vehicles (HFCVs), combined with global efforts to reduce carbon emissions, has accelerated the development of EV charging and hydrogen refueling stations. In response to this demand, this paper introduces the concept of Multi-Functional Charging Station (MFCS) that integrates power generation, EV charging, battery swapping, and hydrogen refueling. A comprehensive operational model is developed for the MFCS that couples electricity and hydrogen conversion and storage technologies to enhance infrastructure utilization and improve overall system efficiency. The model also considers multiple revenue streams, including participation in energy and ancillary markets. To validate the effectiveness of the proposed model and evaluate its performance, a series of numerical experiments are conducted with different charger numbers, different electricity purchase limits, and different charger allocations. Numerical results demonstrate that shared charger configurations can lead to 8.11 % improvement in operational profit by improving resource utilization and reducing the number of depleted batteries at the end of operations compared to allocated charger setups. By varying the number of chargers, sensitivity analysis identifies diminishing marginal returns beyond about 45 chargers, suggesting it as an optimal sizing point under current settings. The integration of electricity and hydrogen conversion is also explored under scenarios with limited external electricity purchases. In conclusion, these findings indicate that optimizing charger allocation and energy management can significantly enhance station productivity and profitability, ultimately supporting the broader adoption of electrified and hydrogen-based transportation solutions.

Charging station↗

DOE OSTI2025

A curvature-based mechanism for the spontaneous emission of electromagnetic radiation at curved plasma–vacuum interfaces

In this work, we derive from first principles the dispersion relation for electromagnetic radiation that is spontaneously emitted at a 2D curved plasma–vacuum interface in the absence of both background and external electric and magnetic fields. This plasma geometry is motivated by plasmas generated through hypervelocity impact projectiles—an area that is of importance to satellite safety in the aerospace community, and may be of importance for the developing study of projectile impact-driven inertial confinement fusion. We find spontaneous radiation occurs when thermal surface waves propagate along the edge of a circular plasma, with a frequency lower than the plasma frequency and spatial decay proportional to 1/r. This is distinct from more well-known mechanisms of radiation, in which the radiation frequency is greater than or equal to the plasma frequency. This theory is validated against 2D, electromagnetic particle-in-cell simulations and shows reasonable agreement. Finally, we discuss the effects of plasma geometry and attribute the radiation generation mechanism to the acceleration of charged surface waves as they traverse azimuthally along a curved path.

Lau, Raymond (ORCID:0000000299318101)↗

DOE OSTI2024

Biosensors for the detection of chorismate and cis,cis -muconic acid in Corynebacterium glutamicum

Abstract Corynebacterium glutamicum ATCC 13032 is a promising microbial chassis for industrial production of valuable compounds, including aromatic amino acids derived from the shikimate pathway. In this work, we developed two whole-cell, transcription factor based fluorescent biosensors to track cis,cis-muconic acid (ccMA) and chorismate in C. glutamicum. Chorismate is a key intermediate in the shikimate pathway from which value-added chemicals can be produced, and a shunt from the shikimate pathway can divert carbon to ccMA, a high value chemical. We transferred a ccMA-inducible transcription factor, CatM, from Acinetobacter baylyi ADP1 into C. glutamicum and screened a promoter library to isolate variants with high sensitivity and dynamic range to ccMA by providing benzoate, which is converted to ccMA intracellularly. The biosensor also detected exogenously supplied ccMA, suggesting the presence of a putative ccMA transporter in C. glutamicum, though the external ccMA concentration threshold to elicit a response was 100-fold higher than the concentration of benzoate required to do so through intracellular ccMA production. We then developed a chorismate biosensor, in which a chorismate inducible promoter regulated by natively expressed QsuR was optimized to exhibit a dose-dependent response to exogenously supplemented quinate (a chorismate precursor). A chorismate–pyruvate lyase encoding gene, ubiC, was introduced into C. glutamicum to lower the intracellular chorismate pool, which resulted in loss of dose dependence to quinate. Further, a knockout strain that blocked the conversion of quinate to chorismate also resulted in absence of dose dependence to quinate, validating that the chorismate biosensor is specific to intracellular chorismate pool. The ccMA and chorismate biosensors were dually inserted into C. glutamicum to simultaneously detect intracellularly produced chorismate and ccMA. Biosensors, such as those developed in this study, can be applied in C. glutamicum for multiplex sensing to expedite pathway design and optimization through metabolic engineering in this promising chassis organism. One-Sentence Summary High-throughput screening of promoter libraries in Corynebacterium glutamicum to establish transcription factor based biosensors for key metabolic intermediates in shikimate and β-ketoadipate pathways.

59 BASIC BIOLOGICAL SCIENCES↗