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At least 19 records

Recombinant LAC polynucleotides and uses thereof to increase production of C-lignin in plants

The disclosure provides recombinant LAC polynucleotides encoding a laccase (LAC) polypeptide capable of polymerizing caffeyl alcohol, vectors and cells including the recombinant LAC polynucleotide. The disclosure also provides transgenic plants including cells having the recombinant LAC polynucleotide of the present disclosure and methods of increasing production of C-lignin in a plant.

Dixon, Richard A.↗

Genetic switching by the Lac repressor is based on two-state Monod–Wyman–Changeux allostery

High-resolution NMR spectroscopy enabled us to characterize allosteric transitions between various functional states of the dimeric Escherichia coli Lac repressor. In the absence of ligands, the dimer exists in a dynamic equilibrium between DNA-bound and inducer-bound conformations. Binding of either effector shifts this equilibrium toward either bound state. Analysis of the ternary complex between repressor, operator DNA, and inducer shows how adding the inducer results in allosteric changes that disrupt the interdomain contacts between the inducer binding and DNA binding domains and how this in turn leads to destabilization of the hinge helices and release of the Lac repressor from the operator. Based on our data, the allosteric mechanism of the induction process is in full agreement with the well-known Monod–Wyman–Changeux model.

59 BASIC BIOLOGICAL SCIENCES↗

LAC Attachments Modernization

This document details the proposed plan to modernize the existing set of cutting attachments in use with the Liquid Abrasive Cutter (LAC). This effort will leverage the replacement of the LAC power generating unit (PGU), which is to be completed by the end of FY25. The PGU modernization will transition away from a diesel direct drive of a high pressure pump to the use of a variable frequency drive, which will be powered by electricity from the existing generator in use by our DOD customers. The existing PGU and a model of the planned replacement is shown below in Figure 1.

42 ENGINEERING↗

An Engineered Laccase from Fomitiporia mediterranea Accelerates Lignocellulose Degradation

Laccases from white-rot fungi catalyze lignin depolymerization, a critical first step to upgrading lignin to valuable biodiesel fuels and chemicals. In this study, a wildtype laccase from the basidiomycete Fomitiporia mediterranea (Fom_lac) and a variant engineered to have a carbohydrate-binding module (Fom_CBM) were studied for their ability to catalyze cleavage of β-O-4′ ether and C–C bonds in phenolic and non-phenolic lignin dimers using a nanostructure-initiator mass spectrometry-based assay. Fom_lac and Fom_CBM catalyze β-O-4′ ether and C–C bond breaking, with higher activity under acidic conditions (pH < 6). The potential of Fom_lac and Fom_CBM to enhance saccharification yields from untreated and ionic liquid pretreated pine was also investigated. Adding Fom_CBM to mixtures of cellulases and hemicellulases improved sugar yields by 140% on untreated pine and 32% on cholinium lysinate pretreated pine when compared to the inclusion of Fom_lac to the same mixtures. Adding either Fom_lac or Fom_CBM to mixtures of cellulases and hemicellulases effectively accelerates enzymatic hydrolysis, demonstrating its potential applications for lignocellulose valorization. We postulate that additional increases in sugar yields for the Fom_CBM enzyme mixtures were due to Fom_CBM being brought more proximal to lignin through binding to either cellulose or lignin itself.

59 BASIC BIOLOGICAL SCIENCES↗

K2 Optical Emission from OJ 287 and Other γ-Ray Blazars on Hours-to-weeks Timescales from 2014 to 2018

We present second observations by K2 of OJ 287 and seven other γ-ray active galactic nuclei (AGNs) obtained in 2017–2018, second and third observations of the lobe-dominated, steep-spectrum quasar 3C 207, and observations of nine additional blazars not previously observed with K2. The AGNs were observed simultaneously with K2 and the Fermi Large Area Telescope for 51–81 days. Our full sample, observed in 2014–2018, contained 16 BL Lacertae objects (BL Lacs), nine flat-spectrum radio quasars (FSRQs), and four other γ-ray AGNs. Twelve BL Lacs and seven FSRQs exhibited fast, jagged light curves, while four BL Lacs and two FSRQs had slow, smooth light curves. Some objects changed their red-noise character significantly between repeated K2 observations. The optical characteristics of OJ 287 derived from the short-cadence K2 light curves changed between observations made before and after the predicted passage of the suspected secondary supermassive black hole through the accretion disk of the primary supermassive black hole. The average slopes of the periodogram power spectral densities of the BL Lacs' and FSRQs' light curves differed significantly, by ≈12%, with the BL Lac slopes being steeper, and a Kolmogorov–Smirnov test with a p-value of 0.039 indicates that these samples probably come from different populations; however, this result is not as strongly supported by power spectral response (PSRESP) analyses. Differences in the origin of the jets from the ergosphere or accretion disk in these two classes could produce such a disparity, as could different sizes or locations of emission regions within the jets.

79 ASTRONOMY AND ASTROPHYSICS↗

A lipase from Lacticaseibacillus rhamnosus IDCC 3201 with thermostability and pH resistance for use as a detergent additive

Lipases are important biocatalysts and ubiquitous in plants, animals, and microorganisms. The high growth rates of microorganisms with low production costs have enabled the wide application of microbial lipases in detergent, food, and cosmetic industries. Herein, a novel lipase from Lacticaseibacillus rhamnosus IDCC 3201 (Lac-Rh) was isolated and its activity analyzed under a range of reaction conditions to evaluate its potential industrial application. The isolated Lac-Rh showed a molecular weight of 24 kDa and a maximum activity of 3438.5 ± 1.8 U/mg protein at 60 °C and pH 8. Additionally, Lac-Rh retained activity in alkaline conditions and in 10% v/v concentrations of organic solvents, including glycerol and acetone. Interestingly, after pre-incubation in the presence of multiple commercial detergents, Lac-Rh maintained over 80% of its activity and the stains from cotton were successfully removed under a simulated laundry setting. Overall, the purified lipase from L. rhamnosus IDCC 3201 has potential for use as a detergent in industrial applications.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Heterologous expression of Arabidopsis laccase2, laccase4 and peroxidase52 driven under developing xylem specific promoter DX15 improves saccharification in populus

Secondary cell wall holds considerable potential as it has gained immense momentum to replace the lignocellulosic feedstock into fuels. Lignin one of the components of secondary cell wall tightly holds the polysaccharides thereby enhancing the recalcitrance and complexity in the biomass. Laccases (LAC) and peroxidases (PRX) are the major phenyl-oxidases playing key functions during the polymerization of monolignols into lignin. Yet, the functions of laccase and peroxidases gene families remained largely unknown. Hence, the objective of this conducted study is to understand the role of specific LAC and PRX in Populus wood formation and to further investigate how the altered Lac and Prx expression affects biomass recalcitrance and plant growth. This study of heterologous expression of Arabidopsis Lac and Prx genes was conducted in poplar to avoid any otherwise occurring co-suppression mechanism during the homologous overexpression of highly expressed native genes. In the pursuit of optimizing lignocellulosic biomass for biofuel production, the present study focuses on harnessing the enzymatic potential of Arabidopsis thaliana Laccase2, Laccase4, and Peroxidase52 through heterologous expression. We overexpressed selected Arabidopsis laccase2 (AtLac2), laccase4 (AtLac4), and peroxidase52 (AtPrx52) genes, based on their high transcript expression respective to the differentiating xylem tissues in the stem, in hybrid poplar (cv. 717) expressed under the developing xylem tissue-specific promoter, DX15 characterized the transgenic populus for the investigation of growth phenotypes and recalcitrance efficiency. Bioinformatics analyses conducted on AtLac2 and AtLac4 and AtPrx52, revealed the evolutionary relationship between the laccase gene and peroxidase gene homologs, respectively. Transgenic poplar plant lines overexpressing the AtLac2 gene (AtLac2-OE) showed an increase in plant height without a change in biomass yield as compared to the controls; whereas AtLac4-OE and AtPrx52-OE transgenic lines did not show any such observable growth phenotypes compared to their respective controls. The changes in the levels of lignin content and S/G ratios in the transgenic poplar resulted in a significant increase in the saccharification efficiency as compared to the control plants. Overall, saccharification efficiency was increased by 35–50%, 21–42%, and 8–39% in AtLac2-OE, AtLac4-OE, and AtPrx52-OE transgenic poplar lines, respectively, as compared to their controls. Moreover, the bioengineered plants maintained normal growth and development, underscoring the feasibility of this approach for biomass improvement without compromising overall plant fitness. This study also sheds light on the potential of exploiting regulatory elements of DX15 to drive targeted expression of lignin-modifying enzymes, thereby providing a promising avenue for tailoring biomass for improved biofuel production. These findings contribute to the growing body of knowledge in synthetic biology and plant biotechnology, offering a sustainable solution to address the challenges associated with lignocellulosic biomass recalcitrance.

09 BIOMASS FUELS↗

The Effect of Physically Based Ice Radiative Processes on Greenland Ice Sheet Albedo and Surface Mass Balance in E3SM

Abstract A significant portion of surface melt on the Greenland Ice Sheet (GrIS) is due to dark ice regions in the ablation zone, where solar absorption is influenced by the physical properties of the ice, light absorbing constituents (LACs), and the overlying crustal surface or melt ponds. Earth system models (ESMs) typically prescribe the albedo of ice surfaces as a constant value in the visible and near‐infrared spectral regions. This work advances ESM ice radiative transfer modeling by (a) incorporating a physically based radiative transfer model (SNow, ICe and Aerosol Radiation model Adding‐Doubling Version 4; SNICAR‐ADv4) into the Energy Exascale Earth System Model (E3SM), (b) determining spatially and temporally varying bare ice physical properties over the GrIS ablation zone from satellite observations to inform SNICAR‐ADv4, and (c) assessing the impacts on simulated GrIS albedo and surface mass balance associated with modeling of more realistic bare ice albedo. GrIS‐wide bare ice albedo in E3SMv2 is overestimated by ∼4% in the visible and ∼7% in the near‐infrared wavelengths compared to the Moderate Resolution Imaging Spectroradiometer. Our bare ice physical property retrieval method found that LACs, ice crustal surfaces, and melt ponds reduce visible albedo by 30% in the bare ice region of the GrIS ablation zone. The realistic bare ice albedo reduces surface mass balance by ∼145 Gt, or 0.4 mm of sea‐level equivalent between 2000 and 2021 compared to the default E3SM. This work highlights the importance of simulating bare ice albedo accurately and realistically to improve our ability to quantify changes in the GrIS surface mass and radiative energy budgets.

58 GEOSCIENCES↗

Spectroscopy of a sample of unidentified gamma-ray Fermi sources

ABSTRACT The fourth-DR3 version (4FGL-DR3) of the Fermi-LAT catalogue of γ-ray sources contains ∼1000 objects at a galactic latitude |b| > 10° that are not identified with an optical counterpart. We performed a systematic study of these sources, focusing on 190 objects that have a unique X-ray counterpart in the available Swift/XRT observations. Optical counterparts were then selected, and optical spectra were found in the literature for 33 sources. We found that 21 could be classified as BL Lac objects. Among these, we were able to provide the redshift for eight of them, while for two others we established a lower limit to the redshift by detecting intervening absorption. The other 12 objects display optical spectra with prominent emission lines (0.036 < z < 1.65). These spectra are characterized by both broad and narrow emission lines, with the exception of three sources. One of these displays only broad emission lines, while the other two exclusively exhibit narrow lines. On the basis of the radio/optical flux ratio, all BL Lac objects in this study are radio-loud. Four sources out of the 12 with prominent emission lines can be classified as radio-loud, while at least five of the 12 sources with prominent lines are radio-quiet. This is somewhat unexpected, given the radio-loudness distribution of 4FGL-associated blazars.

Ulgiati, Alberto (ORCID:0009000736025276)↗

Data for "Discovery, Characterization, and Application of Chromosomal Integration Sites in the Hyperthermophilic Archaeon Sulfolobus islandicus"

Sulfolobus islandicus , an emerging archaeal model organism, offers unique advantages for metabolic engineering and synthetic biology applications owing to its ability to thrive in extreme environments. Although several genetic tools have been established for this organism, the lack of well-characterized chromosomal integration sites has limited its potential as a cellular factory. Here, we systematically identified and characterized 13 artificial CRISPR RNAs targeting eight integration sites in S. islandicus using the CRISPR-COPIES pipeline and a multi-omics-informed computational workflow. We leveraged the endogenous CRISPR-Cas system to integrate the reporter gene lacS and validated heterologous expression through a β-galactosidase assay, revealing significant positional effects. As a proof of concept, we utilized these sites to genetically manipulate lipid ether composition by overexpressing glycerol dibiphytanyl glycerol tetraether (GDGT) ring synthase B (GrsB). This study expands the genetic toolbox for S. islandicus and advances its potential as a robust platform for archaeal synthetic biology and industrial biotechnology.

AI/ML↗

Invasive Plant Species Management Plan for Los Alamos National Laboratory (Rev.3)

Native species are plants and animals that continually occupy a natural range without direct or indirect introduction and/or care by humans. They are adapted to the environmental conditions and processes of the ecosystem in which they reside. A species introduced into a novel ecosystem can either exploit that ecosystem and thrive or be unable to survive in that ecosystem (Hobbs et al. 2006). Alien or non-native species are species that are intentionally or accidentally introduced into a novel ecosystem and are capable of living and propagating within the physical parameters of that ecosystem. Invasive species is a species that is non-native (or alien) to the ecosystem under consideration and a noxious species are those whose introduction causes or is likely to cause economic or environmental harm. The term invasive species is applicable to plants and animals alike; however, this invasive species management plan currently focuses on invasive vegetation. Invasive plant species are usually capable of rapid colonization of disturbed ground, such as after a change in wildfire regime intensity and frequency (Reilly et al. 2020) or anthropogenic ground disturbance (Burke and Grime 1996; Hobbs and Huenneke 1992). Climate change facilitates the spread and establishment of many alien species and creates new opportunities for them to become invasive (Turbelin and Catford 2021). As climate change impacts increase in the coming decades, we may see in an increase in invasive species establishment. See a list of definitions of terms pertinent to this document in Appendix A: Definitions. Los Alamos National Laboratory (LANL) hosts populations of non-native and invasive species all typical of the northern New Mexico region (Martin 2004; NMDA 2020). By implementing an invasive species management plan, LANL will have readily accessible management strategies for invasive species that are found on-site. The benefits of managing invasive species include a decrease in wildland fire risks, an increase in soil productivity, an increase in (productive or beneficial) wildlife habitat, an increase in water quantity and quality, and the restoration of impacted areas (Burke and Grime 1996; Hobbs and Huenneke 1992; D’Antonio and Hobbie 2005; MacDougall et al. 2013; Reilly et al. 2020). The aforementioned benefits from invasive species management directly enable the LANL mission by ensuring compliance requirements are met and site-wide programs, such as the Vulnerability Assessment and Resilience Plan, are supported for a mutually beneficial outcome. An example for the LANL site specifically, controlling non-native annual plant species, for example, could reduce the costs associated with stabilizing soils during the stormwater pollution prevention compliance process. Roadway and utility right-of-way areas are another place where an integrated vegetation management strategy would promote low growing perennial plants in a way that is mutually beneficial to habitat and the institution through lowered maintenance costs. There is also an economic benefit to managing invasive species. In one nationwide study, invasive plants had an estimated impact cost of $190.45 billion (Fantel-Lepczyk et al. 2022). Investing in preventative measures and surveillance could help to offset future control and management costs of invasive species that have the potential to become established. The State of New Mexico has developed plant species lists and recommendations through the New Mexico Department of Agriculture’s Noxious Weeds Management Act, Article 7D (NM Statute § 76-7D-4 2021); however, effective invasive species management must rely on local knowledge of the site and region. Los Alamos County (LAC) has already compiled a target invasive plant species list and species-specific management objectives (Martin 2004). Therefore, the New Mexico Noxious Weed List and the LAC invasive plant species list, as well as management objectives from those documents, are integrated into LANL’s invasive plant species management plan.

54 ENVIRONMENTAL SCIENCES↗

Liquid Abrasive Cutter Power Generation Unit Requirements

This document details the requirements for the purchase of a power generating unit (PGU) for an updated Liquid Abrasive Cutter (LAC) for use in field conditions. The system will provide high pressure water and will be integrated with existing waterjet cutting attachments to cut various materials. The system will be standalone, requiring only water, and electrical energy. Cutting heads, attachments, and a garnet delivery system are not included in this specification. This system is an evolution of the existing LAC (developed at LLNL). The PGU specified herein will have the ability to operate on electrical power via an electric motor, and accept input power from a variety of sources, such as an engine-driven generator, battery pack, solar, shore power, or micro-grid.

42 ENGINEERING↗

Reconstruction of dynamic scenes based on differences between collected view and synthesized view

A system for generating a 4D representation of a scene in motion given a sinogram collected from the scene while in motion. The system generates, based on scene parameters, an initial 3D representation of the scene indicating linear attenuation coefficients (LACs) of voxels of the scene. The system generates, based on motion parameters, a 4D motion field indicating motion of the scene. The system generates, based on the initial 3D representation and the 4D motion field, a 4D representation of the scene that is a sequence of 3D representations having LACs. The system generates a synthesized sinogram of the scene from the generated 4D representation. The system adjusts the scene parameters and the motion parameters based on differences between the collected sinogram and the synthesized sinogram. The processing is repeated until the differences satisfy a termination criterion.

Kim, Hyojin↗

The Global Climate Action Partnership

The Global Climate Action Partnership (GCAP; formerly the Low Emission Development Strategies Global Partnership) is a worldwide network that accelerates ambitious climate goals and advances implementation in Africa, Asia, and Latin America and the Caribbean (LAC). Our strategic approach emphasizes country-driven implementation actions to achieve resilient, just, and inclusive low-emission and net-zero economies. As an incubator for scalable knowledge and solutions, GCAP is leading the way to equitable, climate-resilient, low-emission development.

African Climate Action Partnership↗

Discovery, characterization, and application of chromosomal integration sites in the hyperthermophilic archaeon Sulfolobus islandicus

Sulfolobus islandicus , an emerging archaeal model organism, offers unique advantages for metabolic engineering and synthetic biology applications owing to its ability to thrive in extreme environments. Although several genetic tools have been established for this organism, the lack of well-characterized chromosomal integration sites has limited its potential as a cellular factory. Here, in this work, we systematically identified and characterized 13 artificial CRISPR RNAs targeting eight integration sites in S. islandicus using the CRISPR-COPIES pipeline and a multi-omics-informed computational workflow. We leveraged the endogenous CRISPR-Cas system to integrate the reporter gene lacS and validated heterologous expression through a β-galactosidase assay, revealing significant positional effects. As a proof of concept, we utilized these sites to genetically manipulate lipid ether composition by overexpressing glycerol dibiphytanyl glycerol tetraether (GDGT) ring synthase B (GrsB). This study expands the genetic toolbox for S. islandicus and advances its potential as a robust platform for archaeal synthetic biology and industrial biotechnology.

59 BASIC BIOLOGICAL SCIENCES↗

Optimized CRISPR Interference System for Investigating Pseudomonas alloputida Genes Involved in Rhizosphere Microbiome Assembly

Pseudomonas alloputida KT2440 (formerly P. putida) has become both a well-known chassis organism for synthetic biology and a model organism for rhizosphere colonization. Here, we describe a CRISPR interference (CRISPRi) system in KT2440 for exploring microbe–microbe interactions in the rhizosphere and for use in industrial systems. Our CRISPRi system features three different promoter systems (XylS/P m , LacI/P lac , and AraC/P BAD ) and a dCas9 codon-optimized for Pseudomonads, all located on a mini-Tn7-based transposon that inserts into a neutral site in the genome. It also includes a suite of pSEVA-derived sgRNA expression vectors, where the expression is driven by synthetic promoters varying in strength. We compare the three promoter systems in terms of how well they can precisely modulate gene expression, and we discuss the impact of environmental factors, such as media choice, on the success of CRISPRi. We demonstrate that CRISPRi is functional in bacteria colonizing the rhizosphere, with repression of essential genes leading to a 10–100-fold reduction in P. alloputida cells per root. Finally, we show that CRISPRi can be used to modulate microbe–microbe interactions. When the gene pvdH is repressed and P. alloputida is unable to produce pyoverdine, it loses its ability to inhibit other microbes in vitro. Furthermore, our design is amendable for future CRISPRi-seq studies and in multispecies microbial communities, with the different promoter systems providing a means to control the level of gene expression in many different environments.

Bacteria↗

Fermi -LAT follow-up observations in seven years of real-time high-energy neutrino alerts

The realtime program for high-energy neutrino track events detected by the IceCube South Pole Neutrino Observatory releases alerts to the astronomical community with the goal of identifying electromagnetic counterparts to astrophysical neutrinos. Gamma-ray observations from theFermi-Large Area Telescope (LAT) enabled the identification of the flaring gamma-ray blazar TXS 0506+056 as a likely counterpart to the neutrino event IC-170922A. By continuously monitoring the gamma-ray sky,Fermi-LAT plays a key role in the identification of candidate counterparts to realtime neutrino alerts. In this paper, we present theFermi-LAT strategy for following up high-energy neutrino alerts applied to seven years of IceCube data. Right after receiving an alert, a search is performed in order to identify gamma-ray activity from known and newly detected sources that are positionally consistent with the neutrino localization. In this work, we study the population of blazars found in coincidence with high-energy neutrinos and compare them to the full population of gamma-ray blazars detected byFermi-LAT. We also evaluate the relationship between the neutrino and gamma-ray luminosities, finding different trends between the two blazar classes BL Lacs and flat-spectrum radio quasars.

Astronomy & Astrophysics↗