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All sourcesNASA NTRSDOE OSTI
NASA NTRS2014

Exploration Systems Development (ESD) Approach to Enterprise Risk Management

The National Aeronautics and Space Administration (NASA) Exploration Systems Development (ESD) Division has implemented an innovative approach to Enterprise Risk Management under a unique governance structure and streamlined integration model. ESD's mission is to design and build the capability to extend human existence to deep space. The Enterprise consists of three Programs: Space Launch System (SLS), Orion, and Ground Systems Development and Operations (GSDO). The SLS is a rocket and launch system that will be capable of powering humans, habitats, and support systems to deep space. Orion will be the first spacecraft in history capable of taking humans to multiple destinations within deep space. GSDO is modernizing Kennedy's spaceport to launch spacecraft built and designed by both NASA and private industry. ESD's approach to Enterprise Risk Management is commensurate with affordability and a streamlined management philosophy. ESD Enterprise Risk Management leverages off of the primary mechanisms for integration within the Enterprise. The Enterprise integration approach emphasizes delegation of authority to manage and execute the majority of cross-program activities and products to the individual Programs, while maintaining the overall responsibility for all cross-program activities at the Division. The intent of the ESD Enterprise Risk Management approach is to improve risk communication, to avoid replication and/or contradictory strategies, and to minimize overhead process burden. This is accomplished by the facilitation and integration of risk information within ESD. The ESD Division risks, Orion risks, SLS risks, and GSDO risks are owned and managed by the applicable Program. When the Programs have shared risks with multiple consequences, they are jointly owned and managed. When a risk is associated with the integrated system that involves more than one Program in condition, consequence, or mitigation plan, it is considered an Exploration Systems Integration (ESI) Risk. An ESI risk may require visibility and risk handling by multiple organizations. The Integrated Risk Working Group (IRWG) is a small team of Risk experts that are responsible for collaborating and communicating best practices. In addition, the forum facilitates proper integration of risks across the Enterprise. The IRWG uses a Continuous Risk Management approach for facilitating the identification, analysis, planning, tracking, and controlling of ESI Risks. The ESD Division, Programs, and Integrated Task Teams identify ESI Risks. The IRWG maintains a set of metrics for understanding Enterprise Risk process and the overall Risk Posture. The team is also actively involved in the modeling of risk for Enterprise Performance Management. With the Enterprise being constrained in Schedule and Budget, and with significant technical complexity, the appropriate use of Risk Management techniques is crucial to the success of the Enterprise. The IRWG achieves this through the modified approach, providing a forum for collaboration on risks that cross boundaries between the separate entities.

Bauder, Stephen P.↗
DOE OSTI2018

Preface: Chemical forensics

Introduction: This virtual special issue stems from 36 presentations given at the Fourth Chemical Forensics Symposium held 3 - 4 April 2017 as part of the 253rd American Chemical Society (ACS) National Meeting & Exposition in San Francisco, CA, USA. Since 2011, the ACS Division of Analytical Chemistry has hosted symposia that review developments and topics arising in chemical forensics. Chemical forensics is a developing scientific discipline that aims to attribute a chemical (or mixture) of interest to its source by the analysis of the chemical itself or associated material constituents to address investigative, legal or intelligence questions. “Source” refers to how, where and when a chemical was handled or produced. Given past and recent attacks with chemical warfare agents (CWAs) and other toxic chemicals there is a strong need for research and development in chemical forensics for the purpose of finding those responsible for chemical attacks. Hence, many of the papers in this issue deal with the source attribution of toxic chemicals that can be used as weapons. However, given the needs, principles and methods that are shared across different application areas of material provenance or authentication, papers that deal with the source attribution of other illicitly used chemicals and materials are included.

Chemical Forensic↗
DOE OSTI2022

Heteroanionic Control of Exemplary Second-Harmonic Generation and Phase Matchability in 1D LiAsS 2– x Se x

The isostructural heteroanionic compounds β-LiAsS 2–x Se x (x = 0, 0.25, 1, 1.75, 2) show a positive correlation between selenium content and second-harmonic response and greatly outperform the industry standard AgGaSe 2 . These materials crystallize in the noncentrosymmetric space group Cc as one-dimensional $\frac{1}{∞}$ [AsQ 2 ] - (Q = S, Se, S/Se) chains consisting of corner-sharing AsQ3 trigonal pyramids with charge-balancing Li + atoms interspersed between the chains. LiAsS 2–x Se x melts congruently for 0 ≤ x ≤ 1.75, but when the Se content exceeds x = 1.75, crystallization is complicated by a phase transition. This behavior is attributed to the β- to α-phase transition present in LiAsSe 2 , which is observed in the Se-rich compositions. The band gap decreases with increasing Se content, starting at 1.63 eV (LiAsS 2 ) and reaching 1.06 eV (β-LiAsSe 2 ). Second-harmonic generation measurements as a function of wavelength on powder samples of β-LiAsS 2–x Se x show that these materials exhibit significantly higher nonlinearity than AgGaSe 2 (d 36 = 33 pm/V), reaching a maximum of 61.2 pm/V for LiAsS 2 . In comparison, single-crystal measurements for LiAsSSe yielded a d eff = 410 pm/V. LiAsSSe, LiAsS 0.25 Se 1.75 , and β-LiAsSe 2 show phase-matching behavior for incident wavelengths exceeding 3 μm. The laser-induced damage thresholds from two-photon absorption processes are on the same order of magnitude as AgGaSe 2 , with S-rich materials slightly outperforming AgGaSe 2 and Se-rich materials slightly underperforming AgGaSe 2 .

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2025

Heat conduction in magnetic insulators via hybridization of acoustic phonons and spin-flip excitations

We present a comprehensive study on the longitudinal magnetothermal transport in a paramagnetic effective spin-1/2 magnetic insulator CsYbSe 2 , by introducing a minimal model requiring only Zeeman splitting and magnetoelastic coupling. We use it to argue that hybridized excitations—formed from acoustic phonons and localized spin-flip-excitations across the Zeeman gap of the crystal electric field ground doublet—are responsible for a nonmonotonic field dependence of longitudinal thermal conductivity. Beyond highlighting a starring role for phonons, our results raise the prospect of universal magnetothermal transport phenomena in paramagnetic insulators that originate from simple features shared across many systems. Published by the American Physical Society 2025

Pocs, Christopher A.↗
NASA NTRS2011

Introduction

It would be hard to argue against the fact that Integrated Computational Materials Engineering (ICME) is a fast growing discipline within material science and engineering. A quick scan of the proceedings from conferences such as Aeromat, Material Science and Technology, and the TMS Annual Meeting clearly shows it. What began a few years ago as one symposium has grown into multiple ICME related symposia at each of these conferences. As encouraging as the number of symposia being offered is the attendance at the symposia. For example, one of the ICME symposia at MS&T 10, the symposium in which this book is based, had five sessions which culminated in a panel discussion that was standing room only. In addition to the large, annual materials science and engineering conferences, smaller 1 to 2-day conferences/workshops sponsored by government agencies (e.g. AFRL and NIST) on specific aspects of ICME and by universities promoting their ICME work are regularly offered. And arguably the most significant news with regards to ICME and conferences is the July 2011 First World Congress on ICME. This five day TMS sponsored conference, specifically focused on ICME with an international advisory board of ICME leaders, shows how far ICME has spread across the globe. Evidence for the growth of ICME can also be found in Academia. The University Materials Council (UMC) is composed of department heads for material science and engineering from major U.S. and Canadian universities. Meeting twice a year to share best practices in order to strengthen both the engineering content [1] and the educational process, the UMC s agenda for their Spring 2010 meeting was dedicated to ICME [2]. This meeting was held in response to the growing awareness that the universities play a major role in the success of ICME and therefore need to develop ICME curriculum in order to meet that need. To aid educators in the development of ICME courses, NSF is funding a "Summer School" on ICME to be held at the University of Michigan in 2011 [3]. Northwestern University recently announced a MS Certificate Program in ICME [4]. Course work for this certificate begins in the Fall of 2011. Other signs that ICME is growing comes from the formation of ICME initiatives from work that did not start off with ICME in mind. One of the committees in ASM International is the Materials Properties Database Committee (MPDC). In the 2010 meeting of the MPDC, based on a study by ASM, the committee decided that it would create an ICME sub-committee in order to determine how ASM can meet the growing needs of the ICME community [5]. In 1999, the Air Force Research Laboratory (AFRL) created a consortium, the Metals Affordability Initiative (MAI), with members from both industry and government with a goal of reducing the cost and time to market of producing metal parts for aerospace applications [6].

Arnold, Steven M.↗
NASA NTRS2015

NASA's Rodent Research Project: Validation of Flight Hardware, Operations and Science Capabilities for Conducting Long Duration Experiments in Space

Research using rodents is an essential tool for advancing biomedical research on Earth and in space. Rodent Research (RR)-1 was conducted to validate flight hardware, operations, and science capabilities that were developed at the NASA Ames Research Center. Twenty C57BL/6J adult female mice were launched on Sept 21, 2014 in a Dragon Capsule (SpaceX-4), then transferred to the ISS for a total time of 21-22 days (10 commercial mice) or 37 (10 validation mice). Tissues collected on-orbit were either rapidly frozen or preserved in RNA later at less than or equal to -80 C (n=2/group) until their return to Earth. Remaining carcasses were rapidly frozen for dissection post-flight. The three controls groups at Kennedy Space Center consisted of: Basal mice euthanized at the time of launch, Vivarium controls, housed in standard cages, and Ground Controls (GC), housed in flight hardware within an environmental chamber. FLT mice appeared more physically active on-orbit than GC, and behavior analysis are in progress. Upon return to Earth, there were no differences in body weights between FLT and GC at the end of the 37 days in space. RNA was of high quality (RIN greater than 8.5). Liver enzyme activity levels of FLT mice and all control mice were similar in magnitude to those of the samples that were optimally processed in the laboratory. Liver samples collected from the intact frozen FLT carcasses had RNA RIN of 7.27 +/- 0.52, which was lower than that of the samples processed on-orbit, but similar to those obtained from the control group intact carcasses. Nonetheless, the RNA samples from the intact carcasses were acceptable for the most demanding transcriptomic analyses. Adrenal glands, thymus and spleen (organs associated with stress response) showed no significant difference in weights between FLT and GC. Enzymatic activity was also not significantly different. Over 3,000 tissues collected from the four groups of mice have become available for the Biospecimen Sharing Program. Together, these validation flight findings demonstrate the capability to support long-duration RR on the ISS to achieve both basic science and biomedical objectives.

rodents↗
NASA NTRS2017

Unobtrusive Software and System Health Management with R2U2 on a Parallel MIMD Coprocessor

Dynamic monitoring of software and system health of a complex cyber-physical system requires observers that continuously monitor variables of the embedded software in order to detect anomalies and reason about root causes. There exists a variety of techniques for code instrumentation, but instrumentation might change runtime behavior and could require costly software re-certification. In this paper, we present R2U2E, a novel realization of our real-time, Realizable, Responsive, and Unobtrusive Unit (R2U2). The R2U2E observers are executed in parallel on a dedicated 16-core EPIPHANY co-processor, thereby avoiding additional computational overhead to the system under observation. A DMA-based shared memory access architecture allows R2U2E to operate without any code instrumentation or program interference.

Schumann, Johann↗
NASA NTRSDate not supplied

Microbial Responses to Radiation at the Single-Cell Level

Exploring the biological effects of deep space radiation is crucial for developing protective protocols for humans traveling beyond Earth’s magnetosphere. Saccharomyces cerevisiae, or budding yeast, is an ideal model organism to study the effects of radiation on eukaryotes as it shares several pathways with humans, including DNA repair mechanisms. Here, we used time-lapse imaging of colony growth on agar plates to monitor the effects of radiation exposure on individual yeast cells and their progeny.

Paola J. Maldonado Martinez↗
DOE OSTI2016

Completing the Cleanup of the US Department of Energy's East Tennessee Technology Park - Four Years Early - 16067

On August 1, 2011, URS - CH2M Oak Ridge LLC (UCOR) began its $2.6 billion cleanup of the East Tennessee Technology Park (ETTP), the former Oak Ridge Gaseous Diffusion Plant, located on the U.S. Department of Energy's (DOE) Oak Ridge Reservation (ORR) in Tennessee. Under the contract, the scope of work to be completed includes demolition of all gaseous diffusion buildings, namely K-25, K-31 and K-27, as well as other facilities such as the Toxic Substance Control Act (TSCA) Incinerator, the K-1200 Centrifuge Complex and the K-1037 Barrier Plant. Additionally, UCOR will remove contaminated soils and treat contaminated groundwater. Waste generated by these cleanup activities will be disposed at onsite as well as offsite facilities. UCOR will also continue to manage hundreds of the DOE Oak Ridge Office of Environmental Management (OREM)-owned facilities across the ORR. Due largely to funding challenges early in the project, the DOE estimated in their October 2013 Program Plan 2014-2024 that ETTP cleanup would not be achieved until 2024, four years later than originally scheduled in UCOR's contract with the DOE. In November 2014, the DOE Manager for OREM, Susan Cange, publicly shared her 'vision' for ETTP cleanup by 2020, four years earlier than the DOE's current plan. As part of Vision 2020, all gaseous diffusion buildings would be demolished by the end of 2016. In response to Ms. Cange's vision, UCOR published a Strategic Plan describing how ETTP could be cleaned up by 2020, provided that a number of funding and programmatic and technical challenges could be overcome. UCOR is in the midst of implementing its Strategic Plan, and progress is being made to that end. Since assuming its responsibilities as the ETTP cleanup contractor, UCOR has completed its life-cycle Performance Measurement Baseline (PMB); received its Earned Value Management System (EVMS) certification; completed the D and D of the 44-acre K-25 and 17-acre K-31 gaseous diffusion buildings; completed the K-1070- B Burial Ground remediation project; completed RCRA closure of the TSCA Incinerator; and disposed of more than 541,411 cubic yards of cleanup waste while managing the on-site Environmental Management Waste Management Facility (EMWMF). In 2015, UCOR achieved safety 'Star Status' under the DOE's Voluntary Protection Program (VPP); completed the demolition of the K-31 gaseous diffusion building; advanced the characterization and deactivation of the K-27 gaseous diffusion building, readying the building for the start of demolition in 2016; and began characterization activities in the K-1037 Barrier Plant as well as a number of facilities in the Poplar Creek Area of the ETTP Site. Project performance through December 31, 2015, is as follows: Cost Performance Index - 1.12, Schedule Performance Index - 1.02. Since safety is the foundation of all cleanup work, UCOR's safety record goes hand-in-hand with its project performance. Through calendar year 2015, UCOR's recordable injury rate was 0.94 occurrences per 200,000 work hours, and the day away case rate was 0.31. These rates are both below the DOE and UCOR goals. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2019

International Government Collaboration in ISD (In Situ Decommissioning/In Situ Disposal) - 19261

In situ decommissioning (ISD) is considered by IAEA as the permanent entombment of a legacy facility in 'a structurally long lived material' (e.g. concrete) and is considered a solution only under 'exceptional circumstances' (e.g. following a severe accident). However, this definition of ISD as 'only' entombment is not universally accepted; the approach has been implemented as a decommissioning approach by the US Dept of Energy in the past decade at the Savannah River Site (P and R reactors)2,3, the Hanford site (U Canyon) and at the Idaho site (CP601, CP640 and CP633). The application of ISD outside of the United States has been much less common for large facilities although its use for underground substructures (e.g. discarded pipelines) is commonplace. Proponents of ISD consider it beneficial since it eliminates the need to remove, package and transport contaminated equipment from a nuclear legacy site to a disposal facility and it maximizes the use of an existing nuclear site footprint by consolidating equipment in one place. ISD is considered to offer major safety benefits to both workers and the public, as well as major cost and schedule benefits to Government. Opponents of the approach cite the fact that, following ISD, the decommissioned facility has effectively become a long-term waste disposal site and that regulations pertaining to long term waste disposal have been circumvented. Given the increasing pressure on environmental cleanup budgets and schedules around the world, countries such as the United Kingdom, Canada and Japan are considering ISD in its various forms as a potential path forward for decommissioning certain facilities in lieu of the more traditional approaches of either deferred decommissioning or immediate decontamination/demolition. To address this emerging area of decommissioning, a Governmental Technical Information Exchange meeting was held in December 2017 at the DOE Savannah River Site in Aiken, SC, to bring together responsible parties from the US Dept of Energy (DOE), Atomic Energy of Canada Ltd (AECL), the UK Nuclear Decommissioning Authority (NDA) and the Japanese Atomic Energy Agency (JAEA) and those countries' respective regulatory agencies to share information about respective 'best practices' and challenges with ISD. Representatives from organizations such as IAEA and US Nuclear Regulatory Commission (US NRC) also participated to discuss their organizations' perspectives and position on ISD. Following the meeting, a series of Working Groups were established which have continued the dialogue in three key areas 5; - ISD-Defined: Identify and share case-specific ISD solutions which allows for the full spectrum of needs and which demonstrate that 'entombment' is not the only definition of 'ISD'. - Regulatory - Share and discuss regulatory concerns and challenges with ISD; - Benchmarking - Share information to support informed and transparent decision-making in ISD. This paper discusses the plans for ISD in some of the countries involved in the December meeting and provides an update on the topics being discussed, and progress being made, by the Working Groups that were formed at the meeting. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
NASA NTRS2020

Stereo Combining NASA Technologies and Partnerships to Transform Current-Day Emergency Response Operations

STEReO brings together several technologies in Unmanned Aircraft Systems (UAS) Traffic Management (UTM), Autonomy, Communications, Human Factors, and Domain Expertise & Tools, aimed at providing scalability and flexibility, as well as operational resiliency to dynamic changes during a disaster event. Some of the concepts STEReO explores are: collaborative tools to ingest remote sensing information and distribute a common mission operating picture, apply ad-hoc communication networks to facilitate timely information sharing and communication of changes, vehicle-to-vehicle and onboard autonomy technologies ensure the safety and resiliency of operations, and apply NASA’s UAS traffic management system (UTM) as a public safety UAS Service Supplier (USS) to access and coordinate use of the airspace by both manned and unmanned operations. The potential benefits of STEReO include: standardized, cross-platform communication means increased interoperability and ease of cooperation/collaboration, increased situation awareness and common operating picture allow for earlier detection and decision making, and scalable to size and complexity of environment, operations, and mission objectives. This presentation gives an overview of the STEReO project and introduces a stakeholder workshop as a three-day activity to solicit input from the community of emergency response operators and related industry representatives.

Mercer, Joey S.↗
DOE OSTI2025

Cryogenic optical beam steering for superconducting device calibration

We have developed a calibration system based on a micro-electromechanical systems (MEMS) mirror that is capable of delivering an optical beam over a wavelength range of 180 -- 2000 nm (0.62 -- 6.89 eV) in a sub-Kelvin environment. This portable, integrated system can steer the beam over a $\sim$3 cm $\times$ 3 cm area on the surface of any sensor with a precision of $\sim$100 $μ$m, enabling characterization of device response as a function of position. This fills a critical need in the landscape of calibration tools for sub-Kelvin devices, including those used for dark matter detection and quantum computing. These communities have a shared goal of understanding the impact of ionizing radiation on device performance, which can be pursued with our system. This paper describes the design of the first-generation calibration system and the results from successfully testing its performance at room temperature and 20 mK.

72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS↗
DOE OSTI2014

An atomic-resolution view of neofunctionalization in the evolution of apicomplexan lactate dehydrogenases

Malate and lactate dehydrogenases (MDH and LDH) are homologous, core metabolic enzymes that share a fold and catalytic mechanism yet possess strict specificity for their substrates. In the Apicomplexa, convergent evolution of an unusual LDH from MDH produced a difference in specificity exceeding 12 orders of magnitude. The mechanisms responsible for this extraordinary functional shift are currently unknown. Using ancestral protein resurrection, we find that specificity evolved in apicomplexan LDHs by classic neofunctionalization characterized by long-range epistasis, a promiscuous intermediate, and few gain-of-function mutations of large effect. In canonical MDHs and LDHs, a single residue in the active-site loop governs substrate specificity: Arg102 in MDHs and Gln102 in LDHs. During the evolution of the apicomplexan LDH, however, specificity switched via an insertion that shifted the position and identity of this ‘specificity residue’ to Trp107f. Residues far from the active site also determine specificity, as shown by the crystal structures of three ancestral proteins bracketing the key duplication event. This work provides an unprecedented atomic-resolution view of evolutionary trajectories creating a nascent enzymatic function.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1986

The effects of voice and manual control mode on dual task performance

Two fundamental principles of human performance, compatibility and resource competition, are combined with two structural dichotomies in the human information processing system, manual versus voice output, and left versus right cerebral hemisphere, in order to predict the optimum combination of voice and manual control with either hand, for time-sharing performance of a dicrete and continuous task. Eight right handed male subjected performed a discrete first-order tracking task, time-shared with an auditorily presented Sternberg Memory Search Task. Each task could be controlled by voice, or by the left or right hand, in all possible combinations except for a dual voice mode. When performance was analyzed in terms of a dual-task decrement from single task control conditions, the following variables influenced time-sharing efficiency in diminishing order of magnitude, (1) the modality of control, (discrete manual control of tracking was superior to discrete voice control of tracking and the converse was true with the memory search task), (2) response competition, (performance was degraded when both tasks were responded manually), (3) hemispheric competition, (performance degraded whenever two tasks were controlled by the left hemisphere) (i.e., voice or right handed control). The results confirm the value of predictive models invoice control implementation.

Wickens, C. D.↗
NASA NTRS1989

Automatic speech recognition in air-ground data link

In the present air traffic system, information presented to the transport aircraft cockpit crew may originate from a variety of sources and may be presented to the crew in visual or aural form, either through cockpit instrument displays or, most often, through voice communication. Voice radio communications are the most error prone method for air-ground data link. Voice messages can be misstated or misunderstood and radio frequency congestion can delay or obscure important messages. To prevent proliferation, a multiplexed data link display can be designed to present information from multiple data link sources on a shared cockpit display unit (CDU) or multi-function display (MFD) or some future combination of flight management and data link information. An aural data link which incorporates an automatic speech recognition (ASR) system for crew response offers several advantages over visual displays. The possibility of applying ASR to the air-ground data link was investigated. The first step was to review current efforts in ASR applications in the cockpit and in air traffic control and evaluated their possible data line application. Next, a series of preliminary research questions is to be developed for possible future collaboration.

Armstrong, Herbert B.↗
NASA NTRS1991

National launch system overview with focus on cargo transfer vehicle

As a result of the Augustine Committee's recommendation to the National Space Council, NASA, and the DOD have embarked on a joint program to provide the nation with a new capability for transporting payloads into space. The National Launch System (NLS) consists of a family of modular launch vehicles, combining elements of current launchers (Titan and Shuttle) with newly developed components. This family consists of (1) NLS-1 (a vehicle capable of delivering 80 k to SSF), (2) NLS-2 (a vehicle capable of delivering 50k to LEO), and (3) NLS-3 (a vehicle capable of delivering 20 k to LEO). Management of the program is shared between the two agencies with a Joint Program Office carrying out the level 2 management and integration function while both the NASA and Air Force field organizations are charged with the various development and operational responsibilities.

Buchanan, Harry↗
NASA NTRS1998

Flying Cassini with Virtual Operations Teams

The Cassini Program's challenge is to fly a large, complex mission with a reduced operations budget. A consequence of the reduced budget is elimination of the large, centrally located group traditionally used for uplink operations. Instead, responsibility for completing parts of the uplink function is distributed throughout the Program. A critical strategy employed to handle this challenge is the use of Virtual Uplink Operations Teams. A Virtual Team is comprised of a group of people with the necessary mix of engineering and science expertise who come together for the purpose of building a specific uplink product. These people are drawn from throughout the Cassini Program and participate across a large geographical area (from Germany to the West coast of the USA), covering ten time zones. The participants will often split their time between participating in the Virtual Team and accomplishing their core responsibilities, requiring significant planning and time management. When the particular uplink product task is complete, the Virtual Team disbands and the members turn back to their home organization element for future work assignments. This time-sharing of employees is used on Cassini to build mission planning products, via the Mission Planning Virtual Team, and sequencing products and monitoring of the sequence execution, via the Sequence Virtual Team. This challenging, multitasking approach allows efficient use of personnel in a resource constrained environment.

Dodd, Suzanne↗
DOE OSTI2017

Functional Characterization of Four Putative δ1-Pyrroline-5-Carboxylate Reductases from Bacillus subtilis

In most living organisms, the amino acid proline is synthesized starting from both glutamate and ornithine. In prokaryotes, in the absence of an ornithine cyclodeaminase that has been identified to date only in a small number of soil and plant bacteria, these pathways share the last step, the reduction of δ1-pyrroline-5-carboxylate (P5C) catalyzed by P5C reductase (EC 1.5.1.2). In several species, multiple forms of P5C reductase have been reported, possibly reflecting the dual function of proline. Aside from its common role as a building block of proteins, proline is indeed also involved in the cellular response to osmotic and oxidative stress conditions. Genome analysis of Bacillus subtilis identifies the presence of four genes (ProH, ProI, ProG, and ComER) that, based on bioinformatic and phylogenic studies, were defined as respectively coding a putative P5C reductase. Here we describe the cloning, heterologous expression, functional analysis and small-angle X-ray scattering studies of the four affinity-purified proteins. Results showed that two of them, namely ProI and ComER, lost their catalytic efficiency or underwent subfunctionalization. In the case of ComER, this could be likely explained by the loss of the ability to form a dimer, which has been previously shown to be an essential structural feature of the catalytically active P5C reductase. The properties of the two active enzymes are consistent with a constitutive role for ProG, and suggest that ProH expression may be beneficial to satisfy an increased need for proline.

59 BASIC BIOLOGICAL SCIENCES↗