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At least 19 records
All sourcesNASA NTRSDOE OSTI
DOE OSTI2023

Two disulfide-reducing pathways are required for the maturation of plastid c -type cytochromes in Chlamydomonas reinhardtii

Abstract In plastids, conversion of light energy into ATP relies on cytochrome f, a key electron carrier with a heme covalently attached to a CXXCH motif. Covalent heme attachment requires reduction of the disulfide-bonded CXXCH by CCS5 and CCS4. CCS5 receives electrons from the oxidoreductase CCDA, while CCS4 is a protein of unknown function. In Chlamydomonas reinhardtii, loss of CCS4 or CCS5 yields a partial cytochrome f assembly defect. Here, we report that the ccs4ccs5 double mutant displays a synthetic photosynthetic defect characterized by a complete loss of holocytochrome f assembly. This defect is chemically corrected by reducing agents, confirming the placement of CCS4 and CCS5 in a reducing pathway. CCS4-like proteins occur in the green lineage, and we show that HCF153, a distant ortholog from Arabidopsis thaliana, can substitute for Chlamydomonas CCS4. Dominant suppressor mutations mapping to the CCS4 gene were identified in photosynthetic revertants of the ccs4ccs5 mutants. The suppressor mutations yield changes in the stroma-facing domain of CCS4 that restore holocytochrome f assembly above the residual levels detected in ccs5. Because the CCDA protein accumulation is decreased specifically in the ccs4 mutant, we hypothesize the suppressor mutations enhance the supply of reducing power through CCDA in the absence of CCS5. We discuss the operation of a CCS5-dependent and a CCS5-independent pathway controlling the redox status of the heme-binding cysteines of apocytochrome f.

Genetics & Heredity↗
DOE OSTI2022

A Unified Testing Platform to Mature Blockchain Applications for Grid Emulation Environments

Blockchain technology is a relatively novel technology that can be used to develop more decentralized, autonomous and tamper-evident solutions. A feature that can aid Transactive Energy Systems to reach their goals by enabling individual actors to communicate and reach consensus with other participants in a more decentralized fashion. However, technical barriers to evaluate and adopt this type of technology within the electrical domain still exist. To facilitate this task, BLOSEM Unified Testing Platform (UTP), a DOE-sponsored, multi-lab effort intends to accelerate the development of solutions by offering a common set of reusable services that can be used to interconnect existent grid tools with blockchain services. UTP is intended to serve as development platform that can provide application engineers with the technical means to evaluate potential blockchain solutions, by enabling them to concentrate on the actual application functionalities while at the same time abstracting the connectivity and performance measurement tasks. The use of BLOSEM UTP is further demonstrated by implementing two potential use cases that are intended to validate both the feasibility of implementing these applications as blockchain-based solutions while also demonstrating the features provided by UTP.

blockchain co-simulation↗
DOE OSTI2022

Temporal partitioning of hatching, maturation, and surface activity by reptiles in Florida longleaf pine-wiregrass sandhills

Temporal partitioning of life history traits among syntopic reptiles can facilitate co-occurrence, but may be influenced by environmental factors and evolutionary history. We used 24 years of continuous capture data in the Florida sandhills to evaluate the timing and duration of hatching, maturation and/or surface activity for ten reptile species, spanning multiple clutch strategies, taxonomic relationships, and habits. We hypothesised: i) species would differ in seasonal timing of hatching and maturation; ii) hatching and maturation periods would be more seasonally-synchronised in fossorial than terrestrial or semi-aquatic reptiles; iii) monthly and annual temperature anomalies would be positively related to hatching, maturation, and surface activity anomalies, and iv) groupings of reptiles by clutch strategy, taxonomic relationship, and habit, would explain more variation in the timing and duration of hatching and maturation than species alone. Seasonal timing of response variables varied widely among species. Hatching peaked for > 1 species during most calendar months. Maturation and surface activity periods ranged from aseasonal to highly-seasonal among species. Hatching began 1.5 months earlier and was more prolonged for terrestrial than fossorial species overall. Hatching peaked in early to mid-summer for terrestrial and fossorial species, and winter for the semi-aquatic Kinosternon subrubrum. Terrestrial and fossorial species did not differ in average timing, duration, or overlap of maturation periods; semi-aquatic Liodytes pygaea matured more consistently across all seasons than other species. Monthly temperature anomalies were negatively correlated with monthly maturation for Plestiodon egregius. Annual temperature and precipitation anomalies were related to annual hatching, maturation, and surface activity trends for several species. Taxonomic relationship, habit, and species explained some variation in hatching and maturation timing and duration. Our results illustrate the influence of environment and evolutionary relationships on the timing of important life history traits. Keywords: Age class, Drift fence, Community, Interspecific, Life history

Zoology↗
DOE OSTI2023

Smart manufacturing maturity models and their applicability: a review

The purpose of this paper is to review existing smart manufacturing (SM) maturity models' dimensions and maturity levels to assess their applicability and drawbacks. There are many maturity models available but many of them have not been validated or do not provide a useful guide or tool for applications. This gap creates the need for a review of the existing maturity model's applicability. Nineteen peer-reviewed maturity models related to “Digital Transformation,” “Industry 4.0” or “Smart Manufacturing” were selected based on a systematic literature review and five consulting firm models were selected based on the author's industry knowledge. The chosen models were analyzed to determine 10 categories of dimensions. Then they are assessed on a 1–5 scale for how applicable they are in the 10 categories of dimensions. The five “consulting firm” models have a first-mover advantage, are more widely used in industry and are more applicable, but some require payment, and they lack published details and validation. The 19 “peer reviewed” models are not as widely used, lack awareness in the industry and are not as easy to apply because of no web tool for self-assessment, but they are improving. The categories defined to characterize the models and facilitate comparisons for users include “Information Technology (IT) and Cyber-Physical System (CPS) and Data,” “Strategy and Organization,” “Supply Chain and Logistics,” “Products and Services,” “Culture and Employees,” “Technology and Capabilities,” “Customer and Market,” “Cybersecurity and Risk,” “Leadership and Management” and “Governance and Compliance.” The analyzed maturity models were particularly weak in the areas of cybersecurity, leadership and governance. Researchers and practitioners can use this review with consideration of their specific needs to determine if a maturity model is applicable or if a new model needs to be developed. The review can also aid in the development of maturity models through the discussion of each of the dimension categories. Finally, compared to existing reviews of SM maturity models, this research determines comprehensive dimension categories and focuses on applicability and drawbacks.

42 ENGINEERING↗
DOE OSTI2026

Exploring stages of resilience maturity as communities confront climate risks

Coastal communities are facing increased risks due to climate change, as evidenced by heightened shocks such as hurricanes and wildfires, and stressors including sea-level rise and drought. To confront these risks, many coastal communities are planning and designing their built environments in new ways, with the overall goal of increasing their resilience. However, resilience practices have been widely guided by an event-based timeframe, which may not be sufficient to confront the multiple and overlapping threats expected over the coming decades. To address this concern, this study investigated a long-term perspective on resilience progression, termed resilience maturity. To better understand resilience maturity in practice, this study drew directly on community perspectives, thereby identifying common challenges. Furthermore, variation in resilience maturity across communities provided an opportunity to identify existing successful strategies. Inductive qualitative analysis was used to examine perspectives from 15 interviews with local practitioners across 12 coastal communities. The analysis revealed that communities in early stages of resilience maturity often struggle with local stakeholder alignment and minimal dedicated resilience resources. In contrast, those further along the maturity path often face technology barriers and challenges in aligning regional stakeholders. Few examples of a fully mature resilience culture were found within the communities studied, underscoring systemic barriers to achieving advanced resilience maturity. Overall, these findings supported development of a practical framework for classifying community resilience efforts, identifying common obstacles, and informing localized resilience advancement.

Resilience maturity↗
DOE OSTI2025

The [4Fe-4S] Cluster of HydF Is Essential for [FeFe]-Hydrogenase Maturation

The organometallic H-cluster of the [FeFe]-hydrogenase is assembled in vivo through a complex process requiring the action of three dedicated maturation enzymes, HydG, HydE, and HydF, as well as the aminomethyl-lipoyl-H-protein (H met ) of the glycine cleavage system (GCS). Here we probe the role of HydF and its [4Fe-4S] cluster in [FeFe]-hydrogenase maturation by using a defined semisynthetic approach in which [Fe I 2 (μ-SH) 2 (CO) 4 (CN) 2 ] 2– ([2Fe] E ) is used to bypass HydE and HydG, and GCS components are used in place of cell lysate. We show that inclusion of the iron–sulfur carrier protein NfuA and the high-CO-affinity myoglobin variant Mb H64L provides dramatically improved hydrogenase activities up to 828 μmol/min/mg, equivalent to the best reported activities for Chlamydomonas reinhardtii [FeFe]-hydrogenase isolated from the native organism. Apo-HydF lacking a [4Fe-4S] cluster provides very little hydrogenase activity; however, full maturation is restored with the addition of NfuA, which we demonstrate reconstitutes the [4Fe-4S] cluster of HydF. In addition, a HydF variant lacking a [4Fe-4S] cluster by changing two cysteine ligands to alanine is completely unable to support either semisynthetic maturation using [2Fe] E , or full maturation using HydG and HydE, even in the presence of NfuA, demonstrating that the HydF [4Fe-4S] cluster is absolutely essential for [FeFe]-hydrogenase maturation. The possibility that the HydF [4Fe-4S] cluster plays a role in direct binding of [2Fe] E is negated by our results with the HydF D311C variant, which demonstrate that the labile Asp311 cluster ligand is not essential for [2Fe] E binding and HydA maturation. We therefore conclude that [2Fe]E binds HydF adjacent to, but not directly coordinated to, the [4Fe-4S] cluster. The HydF [4Fe-4S] cluster is proposed to be essential due to its impact on the [2Fe] E binding orientation and the ability of the HydF/[2Fe] E complex to form productive interactions with H met or the H met /T-protein complex during DTMA ligand biosynthesis.

cluster chemistry↗
DOE OSTI2023

Novel Earned Value Management System Maturity Framework and Its Relation to Project Performance

Earned value management systems (EVMS) are used by industry practitioners to successfully manage projects and programs; however, there has been no past study to characterize and evaluate the impact of EVMS maturity on project performance. This paper introduces a novel framework, developed to measure the EVMS maturity, composed of 56 attributes arrayed across ten subprocesses, which together make up an integrated project/program management system. This paper describes how the framework was conceived and developed, based on literature review and data collected from industry workshops. Then, it examines the relationship between EVMS maturity and ten project performance metrics through statistical analyses. The study analyzed project data from 35 projects and programs worth over $21.8 billion in total cost. For this sample, results showed statistically significant differences between projects with high and low EVMS maturity; on average, projects exhibiting high EVMS maturity have 54% less cost overruns and 19% less change orders versus their performance measurement baseline. Higher maturity projects and programs also achieve better customer satisfaction, meet business objectives more often, and are more often in compliance with EVMS standards and guidelines. This paper contributes to the body of knowledge by providing a practical and novel framework for assessing the maturity of earned value management systems. Practitioners can use the framework to improve their integrated project and program management.

99 GENERAL AND MISCELLANEOUS↗
DOE OSTI2025

In vitro maturation of fully active [FeFe]-hydrogenase in a defined system including the iron carrier NfuA

The [FeFe]-hydrogenase employs an active-site 6Fe H-cluster to catalyze the reversible reduction of protons to H 2 . A [4Fe-4S] subcluster of the H-cluster is synthesized by housekeeping iron-sulfur cluster assembly machinery, and then dedicated hydrogenase maturation enzymes, together with components of the glycine cleavage system, build and deliver a [2Fe] subcluster to generate the full H-cluster. Here, we report that theEscherichia coliiron-sulfur carrier protein NfuA supports in vitro maturation of fully active [FeFe]-hydrogenase, with H 2 production rates comparable to that of the in vivo-maturedChlamydomonas reinhardtii[FeFe]-hydrogenase (CrHydA). Inclusion of NfuA in the in vitro maturation process improves its efficacy by delivering the iron essential for formation of the [Fe II (cys)(CN)(CO) 2 ] – synthon at the dangler iron site of the HydG auxiliary cluster. NfuA serves an additional role in reconstituting and maintaining the catalytically essential iron-sulfur clusters on the maturase enzymes HydE, HydF, and HydG. Further inclusion of a high CO affinity myoglobin variant (Mb H64L ) sequesters free CO generated during the maturation process, minimizing formation of the CO-inhibited H ox -CO enzyme state, significantly increasing hydrogenase activity. The addition of NfuA and Mb H64L to the fully defined maturation system thus results in an in vitro [FeFe]-hydrogenase maturation system that generates highly active enzyme while providing insights into factors important to in vivo maturation.

Science & Technology - Other Topics↗
DOE OSTI2025

Secretory stimuli distinctly regulate insulin secretory granule maturation through structural remodeling

Insulin secretory granule (ISG) maturation is a crucial aspect of insulin secretion and glucose homeostasis. The regulation of this maturation remains poorly understood, especially how secretory stimuli affect ISG maturity and subcellular localization. In this study, we used soft X-ray tomography (SXT) to quantitatively map ISG morphology, density, and location in single INS-1E and mouse pancreatic β cells under the effect of various secretory stimuli. We found that the activation of glucokinase (GK), gastric inhibitory polypeptide receptor (GIPR), glucagon-like peptide-1 receptor (GLP-1R), and G protein-coupled receptor 40 (GPR40) promotes ISG maturation. Each stimulus induces unique structural remodeling in ISGs, by altering size and density, depending on the specific signaling cascades activated. These distinct ISG subpopulations mobilize and redistribute in the cell, altering the overall cellular structural organization. Our results provide insight into how current diabetes and obesity therapies impact ISG maturation and may inform the development of future treatments that target maturation specifically.

insulin granule maturation↗
DOE OSTI2024

Ad-Mat: Adaptations of Mature Manufacturing Strategies for Accelerated Redox Flow Battery Deployment

The concept of the Ad-Mat approach is to leverage existing adjacent markets across a broad scope of technologies in order to reduce the manufacturing learning curve and ultimately accelerate redox flow battery (RFB) deployment at scale. Lithium-ion batteries (LIBs) are currently the dominant energy storage technology, and they came to technological maturity under unique market conditions when there was no meaningful competition in the consumer electronic and electric vehicle (EV) space. Today, alternative chemistries that may be technologically better-suited for long-duration storage applications are experiencing a high barrier to entry. This is in large part due to the substantial bias towards the scaled-up production and supply chain that now exists for LIBs. In the case of RFBs in particular, numerous analyses have suggested that RFBs should theoretically have a much lower system cost than LIBs - however, this relies on a mature and competitive manufacturing landscape, which has been extremely challenging to achieve for both flow batteries and other LIB competitors. At the moment, LIB alternatives tend to have isolated small-scale manufacturing pathways, which preclude the economies of scale that would be required to compete with the mature LIB industry. In the present state of the industry, niche manufacturing tools and approaches have evolved to support each alternative technology, such that there is substantial replication and duplication in effort. Continuing to pursue a strategy of isolated manufacturing processes/approaches for each LIB-alternative may never allow for at-scale deployment. In order for RFBs to meaningfully compete with LIBs in the realm of LDES, a new disruptive approach based on cross-industry learning and coordination is needed - and this is exactly what our Ad-Mat concept aims to tackle. In this re-envisioned manufacturing landscape, tools and processes from mature industries can be adapted and deployed across the range of alternative energy storage technologies. Adapting tools, equipment, processes, and industrial learning from mature industries to meet the technological requirements of RFBs would open new markets for existing OEMs in adjacent industries, would prevent unnecessary duplication and re-development, would improve efficiency across the manufacturing chain, and would ultimately support reduced costs and accelerated deployment of RFBs at scale.

adaptive manufacturing↗
DOE OSTI2024

Structural maturation of myofilaments in engineered 3D cardiac microtissues characterized using small angle x-ray scattering

Abstract Understanding the structural and functional development of human-induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs) is essential to engineering cardiac tissue that enables pharmaceutical testing, modeling diseases, and designing therapies. Here we use a method not commonly applied to biological materials, small angle x-ray scattering, to characterize the structural development of hiPSC-CMs within three-dimensional engineered tissues during their preliminary stages of maturation. An x-ray scattering experimental method enables the reliable characterization of the cardiomyocyte myofilament spacing with maturation time. The myofilament lattice spacing monotonically decreases as the tissue matures from its initial post-seeding state over the span of 10 days. Visualization of the spacing at a grid of positions in the tissue provides an approach to characterizing the maturation and organization of cardiomyocyte myofilaments and has the potential to help elucidate mechanisms of pathophysiology, and disease progression, thereby stimulating new biological hypotheses in stem cell engineering.

36 MATERIALS SCIENCE↗
DOE OSTI2023

In vitro maturation of NiSOD reveals a role for cytoplasmic histidine in processing and metalation

The importance of cellular low molecular weight (LMW) ligands in metalloenzyme maturation is largely unexplored. Maturation of NiSOD requires post-translational N-terminal processing of the proenzyme, SodN, by its cognate protease, SodX. Here we provide evidence for the participation of L-histidine in the protease-dependent maturation of Nickel-dependent Superoxide Dismutase (NiSOD) from Streptomyces coelicolor. Furthermore, in vitro studies using purified proteins cloned from S. coelicolor and overexpressed in E. coli support a model where a ternary complex formed between the substrate (SodN), the protease (SodX) and L-Histidine creates a novel Ni-binding site that is capable of the N-terminal processing of SodN and specifically incorporates Ni into the apo-NiSOD product. Thus, L-Histidine serves many of the functions associated with a metallochaperone or, conversely, eliminates the need for a metallochaperone in NiSOD maturation.

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEA↗
DOE OSTI2023

The effect of reducing dietary lipid and food availability on precocious male maturation in Chinook Salmon: A production-scale hatchery experiment

Abstract Objective Age of maturation in Chinook Salmon Oncorhynchus tshawytscha is phenotypically plastic, influenced by both genotype and environmental factors, including the availability and composition of the diet. Salmon hatchery programs often rear fish under accelerated growth regimes using high-lipid diets that can result in earlier age at maturity, including increased prevalence of age-2 males (minijacks). The goal of this investigation was to compare alternative dietary regimes to mitigate for this shift in age at maturity in hatchery-reared Umatilla River fall Chinook Salmon. Methods Juvenile fish were reared at Bonneville Hatchery, Oregon, under four dietary treatments across four replicate brood years. Dietary treatments included two feeding frequencies (standard [fed 7 days/week] and reduced [fed 4 days/week]) and two dietary lipid levels (standard [18%] and reduced [12%]) in a 2 × 2 factorial design. Dietary treatments were applied for approximately 9 months, beginning in March (a month after fry emergence) and lasting until December of the first year, after which all fish were reared on the standard feeding regime (7 days–18%) until the time of release the following spring as yearlings. Result We observed significant interannual variation in the proportion of minijacks produced among dietary treatments. For all brood years, decreasing the feeding frequency from 7 to 4 days/week reduced the proportion minijacks by 35.9%, and lowering dietary lipid from 18% to 12% reduced the proportion minijacks by 30%. The combined effects of reducing the feeding frequency and lowering dietary lipid were additive, reducing the proportion minijacks by 65.5% compared to the standard rearing regime. Growth and energetic indices were monitored throughout and confirmed findings from previous laboratory-based studies indicating that physiological status 10–12 months prior to spawn timing is important for the “decision” to mature. Conclusion Results of this investigation provide useful insights for optimizing rearing regimes for the Umatilla River program and other Chinook Salmon hatchery programs.

Harstad, Deborah L. (ORCID:0000000202676085)↗
DOE OSTI2023

Geocatalytically generated methane from low-maturity coal and shale source rocks at low temperatures (80–120 °C) over 52 months

Geocatalytic methanogenesis has been proposed to contribute to methane generation from low-maturity coal and shale source rocks. This study contributes further evidence for geocatalytic methanogenesis from low-maturity source rocks based on long-term experiments lasting up to 52 months. Samples from the Upper Devonian New Albany Shale (R o 0.54 %) and Springfield Coal No. 2 (Pennsylvanian; R o 0.54 %) were heated in glass tubes at 80, 100, and 120 °C for 52 months. Sample aliquots from the Upper Cretaceous Second White Specks Formation (R o 0.42 %) were heated in gold tubes at 80 and 100 °C for 42 months at elevated hydrostatic pressures of 100 to 300 MPa. The product gases — methane (CH 4 ) and carbon dioxide (CO 2 ) — were collected and quantified, and gas yields were corrected for leakage from imperfectly closed pores in samples during heating. The results show that longer heating produced more CH 4 . The average CH 4 yields from New Albany Shale and Springfield Coal No. 2 are 0.47 and 3.0 μmol CH 4 per gram of total organic carbon (TOC) over 52 months of heating. Elevated hydrostatic pressure caused lower CH 4 yields from the Second White Specks Formation (3.82 to 1.20 μmol g –1 TOC), suggesting that pressure can retard methanogenesis. Maceral type critically controls the methanogenesis potential of low-maturity coal and shale source rocks. Finally, results of this study provide important insights to the origin of natural gas in low-maturity sedimentary basins.

01 COAL, LIGNITE, AND PEAT↗
DOE OSTI2025

Integrative path modeling and QTL mapping identify maturity, stem strength, and cell wall composition driving lettuce resistance to Sclerotinia minor

Lettuce ( Lactuca sativa ) is highly vulnerable to Sclerotinia minor , the pathogen causing lettuce drop. Breeding for resistance is the most effective control strategy; however, full resistance has not been achieved, and current partial resistance sources are often linked with undesirable traits, such as early bolting. This study aimed to unravel the genetic basis of partial resistance to S. minor and its relationship with plant maturity (bolting), stem mechanical strength (SMS), and cell wall composition (CWC) using a recombinant inbred line (RIL) population derived from a cross between the susceptible iceberg cv. ‘Salinas’ and the resistant oil-seed accession PI 251246. Field evaluations indicated that resistance was linked to earlier bolting, stronger stems, and higher pentose content. Path analysis demonstrated that earlier-maturing plants exhibited increased resistance through enhanced SMS and modified CWC, particularly with higher xylose and lower arabinose levels. Further analysis indicated a significant relationship between syringyl lignin content and resistance, especially in plants with varying bolting responses. Three key quantitative trait loci (QTLs) on linkage groups (LG) 2, 6, and 7 were consistently associated with resistance, bolting, and SMS. Importantly, residual QTL analysis revealed that the resistance locus on LG7 acted independently of maturity, suggesting a distinct resistance mechanism. Callose synthase emerged as a key candidate gene within the LG7 resistance QTL, located near - but distinct from - genes associated with plant maturity and flowering. These findings provide valuable insights into decoupling resistance from early bolting, suggesting a pathway for breeding lettuce cultivars with improved disease resistance and delayed bolting.

Lactuca↗
DOE OSTI2023

A genomic region associated with iteroparous spawning phenology is linked with age‐at‐maturity in female steelhead trout

Abstract Age‐at‐maturity and iteroparity are two life history variations of steelhead trout ( Oncorhynchus mykiss ) that are believed to increase population resilience and stability. While repeat‐spawning individuals are thought to have historically made up a substantial portion of the reproductive population in the Columbia River and the majority of females still attempt outmigration as kelts, return rates of repeat‐spawner are low throughout the basin and below 1% for the furthest migrating stocks. Notably, outmigrating adults exhibit variation in rematuration phenology, displaying either “consecutive” (reproduce immediately the following season) or “skip” (delay spawning for future seasons) spawning patterns. Here, we use low coverage whole genome sequencing of consecutive versus skip spawning female Columbia River steelhead from two populations to test for genomic differences between these two iteroparous phenotypes. We identified genomic regions on several chromosomes which were associated with the phenology of iteroparity, including a region on chromosome 25 containing two genes, estradiol receptor beta ( ER β) and glycoprotein hormone beta‐5 ( GPHB5 ), which, in mammals, are estrogen‐sensitive and expressed in reproductive tissues. Allele frequencies in this ER β/ GPHB5 region differed among female steelhead of different age at maturity, but not males. These genes also shared an island of linkage disequilibrium with the SIX6 gene, 600Kbp away on the same chromosome, a region of known association with age‐at‐maturity. These observations contribute to growing evidence that age‐at‐maturity and the phenology of iteroparity are determined by overlapping physiological processes and genetic pathways.

Willis, Stuart↗
DOE OSTI2023

AmeriFlux FLUXNET-1F US-Wi3 Mature hardwood (MHW)

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site US-Wi3 Mature hardwood (MHW). This is the FLUXNET version of the carbon flux data for the site US-Wi3 Mature hardwood (MHW) produced by applying the standard ONEFlux (1F) software. Site Description - The Wisconsin Mature Hardwood site is located in the Washburn Ranger District of the northeastern section of Chequamegon National Forest. A member of the northern coniferous-deciduous biome, surveys from the mid-19th century indicate the region consisted of a mixed stand of red, white, and jack pines. After extensive timber harvesting, wildfires, and farming activity, the region turned into a fragmented mosaic of stands of various ages and composition. As an assemblage, the ten Wisconsin sites are indicative of the successional stages of development in the predominant stand types of a physically homogeneous landscape. The mature hardwood stand represents a typical naturally regenerated second-growth forest, free of anthropogenic disturbances for at least 70 years.

Chen, Jiquan↗
DOE OSTI2022

[FeFe]-Hydrogenase In Vitro Maturation

The [FeFe]-hydrogenase H-cluster is a complex organometallic cofactor whose assembly and installation requires three dedicated accessory proteins referred to as HydE, HydF, and HydG. The roles of these maturases and the precise mechanisms by which they synthesize and insert the H-cluster are not fully understood. This Minireview will focus on new insights into the [FeFe]-hydrogenase maturation process that have been provided by in vitro approaches in which the biosynthetic pathway has been partially or fully reconstructed using semisynthetic and enzyme-based approaches. Specifically, the application of these in vitro, semisynthetic, and fully defined approaches has shed light on the roles of individual maturation enzymes, the nature of H-cluster assembly intermediates, the molecular precursors of H-cluster ligands, and the sequence of steps involved in [FeFe]-hydrogenase maturation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗