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At least 739 records · Page 41

Microbial Competition for Copper: Impacts on Carbon and Nitrogen Cycling

Activities from this DOE sponsored project focused on two general questions: (1) how does competition between microbes for copper affect emissions of greenhouse gasses, especially methane and nitrous oxide? and (2) how does a novel copper-binding compound (methanobactin) made by some methane-oxidizing bacteria bind and reduce metals, as well as what are the regulatory elements involved in controlling methanobactin synthesis? From these efforts, we have conclusively shown that microbial competition for copper does indeed exert significant control on microbial community composition, as well as affects how microbes produce and consume nitrous oxide. We have also developed much better insights into the basis for methanobactin activity, as well as how methanotrophs control its production.

54 ENVIRONMENTAL SCIENCES↗

Extracellular Charge Transport in Microbial Redox Chains: Linking the Living and Non-Living Worlds

The fundamental process of electron transfer (ET) within and between molecules dictates all biological energy conversion strategies, including respiration and photosynthesis. This project resulted in a comprehensive physics-based understanding of the mechanisms and limits of redox networks that mediate and regulate electron transport through microbial metabolic pathways, with special emphasis on environmental microbes that can acquire energy by catalyzing anodic or cathodic reactions on solid-state electrodes (extracellular electron transfer, or EET). By performing electron transfer to/from electrodes, such microbes may be used as biocatalysts for converting the energy stored in diverse chemical fuels to electricity, or vice versa (microbial electrosynthesis), in renewable energy technologies.

59 BASIC BIOLOGICAL SCIENCES↗

Novel Microbial Routes to Synthesize Industrially Significant Precursor Compounds

Ethylene is the most widely employed organic precursor compound in industry. The potential to impact ethylene formation via recently discovered microbial processes is tenable using plentiful CO2 feedstocks. The overall long-term objective of this project was to develop an industrially compatible microbial process to synthesize ethylene in high yields. The key objective of this project was to fully define and initially characterized a recently discovered and genetically regulated anaerobic pathway to produce high levels of ethylene called the Dihydroxyacetone Phosphate - Ethylene Pathway in phototrophic bacteria. This was addressed through the following specific aims: 1. Fully probe the catalytic potential of all enzymes of the DHAP ethylene pathway and determine the regulatory mechanism of DHAP-ethylene pathway gene expression. 2. Discover effective and active ethylene enzymes encoded in cultured and uncultured organisms from anoxic environments. 3.Model the thermodynamics and kinetics of ethylene synthetic pathways to guide engineering efforts in integrating best performing DHAP-ethylene pathway enzymes into model bacteria chassis for enhance ethylene yields. Through this project we discovered the initially missing genetic and enzyme component of the DHAP-ethylene pathway that directly synthesized ethylene and other important industrial compounds like methane and ethane from specific substrates. We uncovered and partially characterized a nitrogenase-like reductase that functions in DHAP-ethylene pathway specifically and in methionine synthesis in general. This nitrogenase-like system is called the Methylthio-Alkane Reductase (MAR) for its ability to cleave volatile organic sulfur compounds into methanethiol (CH3-SH) for methionine synthesis and a hydrocarbon byproduct. Key to the DHAP-ethylene pathway, MAR is the essential enzyme that cleaves 2-methylthioethanol (CH3-S-CH2-CH2-OH) into ethylene. Coordinately, we uncovered that the MAR genes and genes associated with conversion of methanethiol (CH3-SH) to methionine are under genetic control of a LysR Type Transcriptional Regulator called SalR, whose activity is dependent upon the amount of sulfate available to the cell. When sulfate as the preferred sulfur source for cell growth drops below 200 micromolar, SalR become active for expressing the MAR and methionine biosynthesis genes to enable the cell to grow from volatile organic sulfur compounds and make ethylene. Metabolic thermos-kinetic modeling revealed that these MAR reactions for ethylene and other hydrocarbon production are highly thermodynamically favorable and are one of the largest driving forces for ethylene production by the DHAP-ethylene pathway for high ethylene yields. Modeling also indicated that a key aldolase and to a lesser extent an isomerase of the DHAP-ethylene pathway for production of the ethylene precursor, 2-methylthioethanol, also would increase ethylene yields. Through metagenomic mining and gene synthesis by the JGI DNA synthesis program, over 500 aldolase and isomerase homologs were synthesized and screened. From this, variants were uncovered with substantially higher activity that increased ethylene yields 5-fold via the aldolase reaction and 1.5-fold via the isomerase reaction. Each of these elements that increase ethylene production were integrated together via plasmid under appropriate gene promoter elements in the phototrophic bacterium, Rhodospirillum rubrum, resulting in at least 3 orders of magnitude increase in ethylene yield from carbon dioxide feedstock.

10 SYNTHETIC FUELS↗

Effects of Surface Water Fluctuations and Sediment Movement on Hyporheic Zone Biogeochemistry and Microbial Communities

The hyporheic zone (HZ) is the region beneath and adjacent to streams, rivers, and riverine estuaries where the bidirectional exchange of groundwater (GW) and surface water (SW) occurs. The HZ is an integral component of watersheds, a biologically-active zone providing critical control of SW quality and aquatic life. Much prior work has investigated microbially-mediated biogeochemical reactions in the HZ, mainly from a steady-state perspective. Yet key unique aspects of the HZ include 1) proximity to SW fluctuations (e.g., storms, dam operations) which cyclically drive DOC and DO into the bed and banks creating biogeochemical “hot moments,” and 2) continually shifting surface sediment. Both of these controls should substantially influence microbial communities, yet remain poorly understood, and a mechanistic predictive framework does not exist.

54 ENVIRONMENTAL SCIENCES↗

Microbial Analysis of 100-N Borehole Samples: Estimation of Diesel and BTEX Degrading Bacterial Populations

Historical unplanned releases at the UPR-100-N-17 166-N Diesel Oil Supply Line Leak waste site located within the 100-NR-1 OU resulted in diesel fuel contamination of the unsaturated vadose zone and site groundwater. Enhanced in situ bioremediation of the deep vadose zone was the final remedy decision for the 100-NR-1 Operable Unit (OU) and the full-scale bioventing demo began in 2012. For this remedy, oxygen is actively supplied to the deep vadose zone to stimulate aerobic hydrocarbon degrading microorganisms to metabolize petroleum contamination. Degradation rate calculations based on vadose zone respirometry measurements, however, have indicated declining activity over recent years, though the precise reason for this apparent decline has not been determined. In this investigation 100-NR-1 OU sediment cores were analyzed for total microbial biomass, as well as enumeration of diesel and BTEX degrading bacteria to determine if these microbial populations have declined relative to previous measurements taken in 2009.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

MR13A-3183: Microbial and Geochemical Characterization of Groundwater: Implications for Underground Hydrogen Storage Leakage

Underground hydrogen storage (UHS) in geological formations is a key element of the clean energy transition as it enables the decarbonization of the transportation and industrial sectors by decoupling hydrogen production and storage. UHS has many benefits, including low cost, much wider availability, large storage capacity, well-established infrastructure, and increased safety because of geological sealing capabilities. However, the impact of hydrogen (H2) biogeochemical interactions in the presence of subsurface microorganisms is largely neglected from UHS perspectives. These interactions might affect the effectiveness of storage and can even cause H2 to leak into the shallow aquifers. Leakage of H2 into groundwater can change the geochemistry and induce several microbial-driven processes. Microorganisms, such as sulfate-reducers, are naturally abundant in groundwater and consume H2 to produce hydrogen sulfide (H2S), which can contaminate the freshwater drinking groundwater and cause damage to infrastructure. Hydrogen leakage can also trigger microbial reactions responsible for metal mobility, which can impact the water quality. However, the kinetics of these reactions and the temporal impact of hydrogen leakage in groundwater are still unknown. Therefore, a time series hydrogen-groundwater interaction experiment was conducted, and the changes in fluid chemistry and headspace gas composition will be analyzed along with DNA sequencing results to understand the extent and kinetics of biogeochemical reactions that occur if hydrogen leaks into groundwater. In the experiments, Ultra High Purity (UHP) hydrogen gas will be injected into glass vials with groundwater samples, for a designated time period. For each glass-sealed vial, 16S rRNA gene sequencing, IC, ICP-MS, and GC-TCD will be performed. The experiments provide insights into plausible impacts of hydrogen leakage into shallow drinking water aquifers.

Clark, Allison [West Virginia University (WVU)]↗

Characterization and Stimulation of Selenium Reducing Microbial Consortia from Real Flue Gas Desulfurization Wastewater Biological Treatment System

Selenium is released into the environment via anthropogenic activities such as coal combustion. The U.S. Environmental Protection Agency’s Effluent Limitation Guideline (ELG) Rule limits the release of selenium species into U.S. waterways. To meet the ELG limit for selenium, coal-fired power plants must treat the wastewater produced by the flue gas desulfurization (FGD) systems. Selenium removal is of interest because small changes of the concentrations of selenium in the environment can cause detrimental ecological consequences. Biological treatment is used to remediate this wastewater, but little is known about the individual species in the microbial consortium that reduce toxic water-soluble selenium oxyanions to insoluble elemental selenium. To optimize current biological treatment capabilities of this water, the selenium-reducing capabilities must be leveraged. In order to do so, the selenium-reducing bacteria (SeRB) in biological treatment systems must be characterized. This study will be the first to characterize the microbial consortia from a real FGD WW biological treatment system and provide insights on how to better achieve selenium removal via biological treatment. This work will provide insights into the microorganisms predominantly responsible for the reduction of selenium oxyanions in FGD wastewater biological treatment systems, which can be used to better optimize more efficient biological treatment strategies of this industrial waste.

geomicrobiology↗

From viruses to protists: temperature response of the neglected components of microbial controls on peatland nutrient cycling

The response of plant-associated microbial communities to rising temperatures likely plays a key role in global Carbon (C) and Nitrogen (N) biogeochemical cycles. Mosses of the genus Sphagnum, in particular, sequester 25% of all terrestrial C as recalcitrant peat. However, their ability to sequester C is mediated by interactions with microbes that fix N and pass it to the moss to grow in otherwise oligotrophic systems. Two important biotic controls on this processes are likely to respond to rising temperatures –predation by protists and infections by viruses– but whether and how this might occur is not well understood. This proposal addressed these questions using a combination of genomics and mathematical modeling with both field data from whole-ecosystem warming experiments, and laboratory-based microcosm experiments. We have discovered 1) mechanistic links between cell traits and thermal performance in protists, which 2) enabled predictions from populations to ecosystems, 3) we have identified eco-phenotypic feedbacks that determine protist–bacterial predator–prey dynamics, 4) have documented concomitant compositional and trait shifts in protist communities with warming in the field, 5) revealed consistent interactive effects of temperature and nutrients on bacterial and protist communities; 6) characterized the Sphagnum virome, and 7) made modeling predictions on its implications for thermal responses of microbial communities in peatlands globally. This award also substantially strengthened U.S. scientific capacity and workforce development.

Gibert, Jean Philippe [Duke University]↗

Developing a one-step, acid-free microbial process to recover and separate REE (CRADA Final Report)

The project aimed to develop and test microbial strains for increased bioleaching, bioconcentration, and/or bioaccumulation of REEs, and for enhanced separation of REEs from other metals and of individual REEs from one another. The team secured and tested unconventional REE feedstocks to evaluate strains/processes for bioleaching, bioconcentration, and bioaccumulation, including but not limited to wastewater, geothermal brines, red mud, and electronic waste. purified and chemically analyzed REE granules from the microbial platform; determined metabolite and biosynthetic impacts of growth with REEs and the composition of extracellular material(s) produced by platform strains; optimized the growth process with alternative carbon feedstocks to reduce the cost of the fermentation process; and scaled the process from 1 mL to 10 L.

36 MATERIALS SCIENCE↗

Removal of primary nutrient degrading members severely reduces growth of soil microbial communities even when additional degraders are present

Understanding how microorganisms within a soil community interact to support collective respiration and growth remains challenging. Here we used a model substrate, chitin, and a Model Soil Consortium, MSC-2, to investigate how individual members of a microbial community contribute to decomposition and community growth. While MSC-2 can grow using chitin as the sole carbon source, we do not yet know how the growth kinetics or final biomass yields of MSC-2 vary when certain chitin degraders, or other important members, are absent. To characterize specific roles within this representative community, we carried out experiments leaving out members of MSC-2 and measuring biomass yields and CO2 production. We chose two members to iteratively leave out (referred to by genus name): Streptomyces, as it is predicted via gene expression analysis to be a major chitin degrader in the community, and Rhodococcus as it is predicted via species co-abundance analysis to interact with several other members. Our results showed that when MSC-2 lacked Streptomyces, growth and respiration of the community was severely reduced. Removal of either Streptomyces or Rhodococcus led to major changes in abundance for several other species, pointing to a comprehensive shifting of the microbial community when important members are removed as well as alterations in the metabolic profile, especially when Streptomyces was removed. These results show that when keystone, chitin degrading members are removed, other members, even those with the potential to degrade chitin, do not fill the same metabolic niche to promote community growth. In addition, highly connected members may be removed with similar or even increased levels of growth and respiration. Our findings are critical to a better understanding of soil microbiology, specifically in how communities maintain activity when biotic or abiotic factors lead to changes in biodiversity in soil systems.

McClure, Ryan S↗

KBase Narrative - Formation of a constructed microbial community in a nutrient rich environment indicates bacterial interspecific competition

This Narrative is the parent for the three Narratives referenced in "Formation of a constructed microbial community in a nutrient rich environment indicates bacterial interspecific competition." Wang J, Appidi MR, Burdick LH, Abraham PE, Hettich RL, Pelletier DA, Doktycz MJ. 2024. Formation of a constructed microbial community in a nutrient-rich environment indicates bacterial interspecific competition. mSystems. American Society for Microbiology.

Wang, Jia↗

Fermented Foods Microbial Genomes Database

This database contains ~4,300 microbial genomes assembled from diverse fermented foods. These genomes were obtained from a larger set of 13,850 microbial genomes by clustering them at 99% average nucleotide identity (ANI) to create a "species"-representative database.

59 BASIC BIOLOGICAL SCIENCES↗

Corynebacterium glutamicum as an Efficient Omnivorous Microbial Host for the Bioconversion of Lignocellulosic Biomass

Corynebacterium glutamicum has been successfully employed for the industrial production of amino acids and other bioproducts, partially due to its native ability to utilize a wide range of carbon substrates. We demonstrated C. glutamicum as an efficient microbial host for utilizing diverse carbon substrates present in biomass hydrolysates, such as glucose, arabinose, and xylose, in addition to its natural ability to assimilate lignin-derived aromatics. As a case study to demonstrate its bioproduction capabilities, L-lactate was chosen as the primary fermentation end product along with acetate and succinate. C. glutamicum was found to grow well in different aromatics (benzoic acid, cinnamic acid, vanillic acid, and p-coumaric acid) up to a concentration of 40 mM. Besides, 13 C-fingerprinting confirmed that carbon from aromatics enter the primary metabolism via TCA cycle confirming the presence of β-ketoadipate pathway in C. glutamicum . 13 C-fingerprinting in the presence of both glucose and aromatics also revealed coumarate to be the most preferred aromatic by C. glutamicum contributing 74 and 59% of its carbon for the synthesis of glutamate and aspartate respectively. 13 C-fingerprinting also confirmed the activity of ortho-cleavage pathway, anaplerotic pathway, and cataplerotic pathways. Finally, the engineered C. glutamicum strain grew well in biomass hydrolysate containing pentose and hexose sugars and produced L-lactate at a concentration of 47.9 g/L and a yield of 0.639 g/g from sugars with simultaneous utilization of aromatics. Succinate and acetate co-products were produced at concentrations of 8.9 g/L and 3.2 g/L, respectively. Our findings open the door to valorize all the major carbon components of biomass hydrolysate by using C. glutamicum as a microbial host for biomanufacturing.

13C-fingerprinting↗

Godzilla mineral dust and La Soufrière volcanic ash fallout immediately stimulate marine microbial phosphate uptake

During the “Godzilla” dust storm of June 2020, unusually high fluxes of mineral dust traveled across the Atlantic from the Sahara Desert, reaching the Caribbean Basin, Gulf Coast, and southeastern United States. Additionally, an eruption of the La Soufrière volcano on St. Vincent in April 2021 generated substantial ashfall in the southeastern Caribbean. While many studies have analyzed mineral dust’s ability to relieve nutrient limitation of phosphorus (P) in the P-stressed North Atlantic, less is known about the impact of extreme events and other natural aerosols on fluxes of P into seawater and from seawater into marine microbial cells. We quantified P and iron (Fe) content in mineral dust from the Godzilla dust storm and volcanic ash from the La Soufrière eruption collected at Ragged Point, Barbados. We also performed seawater incubations to assess the marine microbial response to aerosol deposition. Using environmentally-relevant concentrations of atmospheric particles for within the ocean’s mixed layer allowed us to draw realistic conclusions about how these deposition events impacted P cycling in situ. Volcanic ash has lower P content than mineral dust, and P in volcanic ash is far less soluble (~1%) than assumed in current atmospheric deposition models. Adding mineral dust and the volcanic ash leachate in concentrations representing different deposition scenarios increased soluble reactive phosphorus (SRP) concentrations in coastal seawater by ~7-32 nM. Phosphate uptake rate was stimulated in coastal seawater after either mineral dust or volcanic ash deposition at aerosol concentrations relevant to the Godzilla dust event, with ash eliciting the fastest uptake rate. Furthermore, high concentrations of both the mineral dust and volcanic ash led to slightly elevated alkaline phosphatase activity (APA) compared to the relevant controls, indicating higher potential for use of dissolved organic phosphorus (DOP) as a P source. Quantifying these aerosols’ impacts on P cycling is a significant step towards achieving a better understanding of their potential roles in relieving nutrient limitation and fueling the biological carbon pump.

54 ENVIRONMENTAL SCIENCES↗

Long-term integrated soil-crop management improves soil microbial community structure to reduce GHG emission and increase yield

Integrated soil-crop management (ISCM) has been shown as an effective strategy to increase efficiency and yield while its soil microbial community structure and function remain unclear. We evaluated changes in soil physicochemical factors, bacterial community structure responses, and the contributions of soil properties and bacterial communities to summer maize-winter wheat yield and GHG emissions through an ISCM experiment [T1 (local smallholder farmers practice system), T2 (improved management system), T3 (high–yield production system), and T4 (optimized management system)], which could provide scientific guidance for sustainable development of soil in summer maize-winter wheat rotation system. The results showed that the optimized ISCM could improve the soil quality, which significantly changed the soil bacterial community structure to reduce GHG emissions and increase yield. The co-occurrence network density of T3 was increased significantly. The Acidobacteria (class) and OM190 (class) were enriched in T2 and T4. The Frankiales (order) and Gaiellales (order) were enriched in T3. However, the changes in different crop growth stages were different. At the wheat jointing stage and maize mature stage, T4 could enhance carbon-related functional groups, such as aromatic hydrocarbon degradation and hydrocarbon degradation, to increase the soil organic carbon content. And at the maize tasseling stage, T4 could enhance nitrogen-related functional groups. And soil bacteria structure and function indirectly affected annual yield and GHG emission. T2 and T4 exhibited a similar soil microbial community. However, the yield and nitrogen use efficiency of T2 were reduced compared to those of T4. The yield of T3 was the highest, but the GHG emission increased and soil pH and nitrogen use efficiency decreased significantly. Therefore, T4 was a suitable management system to improve soil quality and soil bacterial community structure and function to decrease GHG emissions and increase the yield of the summer maize-winter wheat rotation system.

Yu, Ningning↗

Removal of primary nutrient degraders reduces growth of soil microbial communities with genomic redundancy

Understanding how microorganisms within a soil community interact to support collective respiration and growth remains challenging. Here, we used a model substrate, chitin, and a synthetic Model Soil Consortium (MSC-2) to investigate how individual members of a microbial community contribute to decomposition and community growth. While MSC-2 can grow using chitin as the sole carbon source, we do not yet know how the growth kinetics or final biomass yields of MSC-2 vary when certain chitin degraders, or other important members, are absent. To characterize specific roles within this synthetic community, we carried out experiments leaving out members of MSC-2 and measuring biomass yields and CO 2 production. We chose two members to iteratively leave out (referred to by genus name): Streptomyces, as it is predicted via gene expression analysis to be a major chitin degrader in the community, and Rhodococcus as it is predicted via species co-abundance analysis to interact with several other members. Our results showed that when MSC-2 lacked Streptomyces, growth and respiration of the community was severely reduced. Removal of either Streptomyces or Rhodococcus led to major changes in abundance for several other species, pointing to a comprehensive shifting of the microbial community when important members are removed, as well as alterations in the metabolic profile, especially when Streptomyces was lacking. These results show that when keystone, chitin degrading members are removed, other members, even those with the potential to degrade chitin, do not fill the same metabolic niche to promote community growth. In addition, highly connected members may be removed with similar or even increased levels of growth and respiration. Our findings are critical to a better understanding of soil microbiology, specifically in how communities maintain activity when biotic or abiotic factors lead to changes in biodiversity in soil systems.

59 BASIC BIOLOGICAL SCIENCES↗

Microbial communities in the liver and brain are informative for postmortem submersion interval estimation in the late phase of decomposition: A study in mouse cadavers recovered from freshwater

Introduction Bodies recovered from water, especially in the late phase of decomposition, pose difficulties to the investigating authorities. Various methods have been proposed for postmortem submersion interval (PMSI) estimation and drowning identification, but some limitations remain. Many recent studies have proved the value of microbiota succession in viscera for postmortem interval estimation. Nevertheless, the visceral microbiota succession and its application for PMSI estimation and drowning identification require further investigation. Methods In the current study, mouse drowning and CO 2 asphyxia models were developed, and cadavers were immersed in freshwater for 0 to 14 days. Microbial communities in the liver and brain were characterized via 16S rDNA high-throughput sequencing. Results Only livers and brains collected from 5 to 14 days postmortem were qualified for sequencing. There was significant variation between microbiota from liver and brain. Differences in microbiota between the cadavers of mice that had drowned and those only subjected to postmortem submersion decreased over the PMSI. Significant successions in microbial communities were observed among the different subgroups within the late phase of the PMSI in livers and brains. Eighteen taxa in the liver which were mainly related to Clostridium_sensu_stricto and Aeromonas , and 26 taxa in the brain which were mainly belonged to Clostridium_sensu_stricto , Acetobacteroides , and Limnochorda , were selected as potential biomarkers for PMSI estimation based on a random forest algorithm. The PMSI estimation models established yielded accurate prediction results with mean absolute errors ± the standard error of 1.282 ± 0.189 d for the liver and 0.989 ± 0.237 d for the brain. Conclusions The present study provides novel information on visceral postmortem microbiota succession in corpses submerged in freshwater which sheds new light on PMSI estimation based on the liver and brain in forensic practice.

Wang, Linlin↗

Editorial: Structure and mechanism of microbial membrane active transporters

Membrane active transporters play essential roles in microbial physiology. They couple energy transduction to conformational changes that drive translocation of nutrients, substrates and ions, as well as molecular communication. The structure and function of microbial membrane active transporters are highly diverse. Typical examples include the primary active transporters in the ATP-binding cassette (ABC) superfamily (Thomas and Tampé, 2020; Davidson et al., 2008; Locher et al., 2002), the secondary active transporters in the Major Facilitator Superfamily (MFS) (Drew et al., 2021; Kaback and Guan, 2019), and the ligand-gated porins in the TonB-dependent transporter (TBDT) family (Klebba et al., 2021). As structural, proteogenomic, and computational methods advance, active transporters are increasingly recognized as dynamic molecular machines whose mechanisms can now be visualized and modeled with remarkable precision, building on decades of biochemical and biophysical discovery that established the foundations of this field. The transporter studies recruited in this Research Topic provide us with new insights into the field including structure-function of sugar transporters in yeast, structural prediction and classification of ABC complexes in Bacillus subtilis, Type VI Secretion System (T6SS) in Bacteroides fragilis, amino acids uptake in Escherichia coli and bacterial spore germination.

mechanism↗