Fungal laccases have high activity in degrading various persistent organic pollutants. However, using enzymes in solution for water treatment has limitations of nonreusability, short enzyme lifetimes, and high cost of single use. In this study, we developed a new type of biocatalyst by immobilizing fungal laccase on the surface of yeast cells using synthetic biology techniques. The biocatalyst, referred to as surface display laccase (SDL), had an enzyme activity of 104 ± 3 mU/g dry cell (with 2,2-azinobis-3-ethylbenzothiazoline-6-sulfonate (ABTS)). The SDL retained over 90% of the initial enzyme activity after 25 days storage at room temperature, while, in contrast, activity of free laccase declined to 60% of its initial activity. The SDL could be reused with high stability as it retained 74% of initial activity after eight repeated batch reactions. Proof-of-concept evaluations of the effectiveness of SDL in treating contaminants of emerging concern were performed with bisphenol A and sulfamethoxazole. Results from contaminant degradation kinetics and the effects of redox mediator amendment provided insights into the factors affecting the efficacy of the SDL system. This study reports, for the first time, the development of a surface display enzyme biocatalyst as an effective and renewable alternative for treating recalcitrant organic micropollutants.
In large regions of the ocean, low iron availability regulates diatom growth and species composition. Diatom species often vary in their physiological response to iron enrichment, with natural and artificial iron additions in iron‐limited regions of the ocean resulting in large blooms of primarily pennate diatoms. The ability of pennate diatoms to proliferate following pulse iron additions has been partly attributed to their ability to acquire and store excess intracellular iron in the iron storage protein ferritin. Recent transcriptome sequencing of diatoms indicate that some centric diatoms also possess ferritin. Using a combination of physiological and molecular techniques, we examined the iron storage capacities and associated ferritin gene expression in phylogenetically diverse centric and pennate diatoms grown under high and low iron concentrations. There were no systematic differences among ferritin‐containing and non‐containing diatom lineages in their ability to store iron in excess of that needed to support maximum growth rates. An exception, however, was the ferritin‐containing pennate diatom Pseudo‐nitzschia granii, native to iron‐limited waters of the Northeast Pacific Ocean. This species exhibited an exceptionally large luxury iron storage capacity and increased ferritin gene expression at high iron concentrations, supporting a role in long‐term iron storage. By contrast, two other diatoms species that exhibited minimal iron storage capacities contained two distinct ferritin genes where one ferritin gene increased in expression under iron limitation while the second showed no variation with cellular iron status. We conclude that ferritin may serve multiple functional roles that are independent of diatom phylogeny.
The Antrim Shale, located in the Michigan Basin, United States (U.S.), is a major U.S. shale play having produced over 2.5 Trillion Cubic Feet (Tcf) of unconventional shale natural gas as of 2010. The shallow nature of this formation sets it apart from other, more characterized unconventional shale gas plays. The depth of gas production of the Antrim ranges from approximately 150 to 600 m and it is typically vertically drilled, contrary to deeper, horizontally drilled shales. A thorough understanding of the biogeochemistry and microbiology of this complex system will be advantageous for improving well performance, produced water management, and potential biocidal treatment as microbial community composition can vary substantially even among closely spaced wells. In this study, we analyzed produced water collected from nine different wells in the Antrim Shale by investigating the geochemical and microbial community composition of the produced water to gain greater insight into the overall biogeochemistry of this unique shale system. The majority of the wells from this study had high total dissolved solids (TDS) primarily composed of chloride and sodium, averaging 86 804 mg/L with a maximum 116 223 mg/L; however, three of the wells sampled along the northern margin of the basin exhibited significantly lower TDS ranging from 4932 to 6496 mg/L. Our microbial community analysis revealed relatively low abundance within our samples and high variability of the microbial community among the sampled wells. The majority of bacterial sequences were identified within Proteobacteria, Firmicutes, and Actinobacteria phyla and metagenomic sequencing revealed the low presence of Methanobacteriaceae within each sample. We also investigated potential microbial community drivers and found that TDS, sodium, chloride, iodide, bromide, ammonium, potassium, and strontium were significantly correlated with the observed microbial community. The varying geochemical conditions between wells demonstrate different subsurface environmental niches, potentially driving the heterogeneous microbial communities we observed from well to well. This analysis suggests an important relationship between both well location and geochemistry and the observed microbial community that can persist in the reservoir. Continued studies of the Antrim Shale will improve our understanding of the complex interdependencies of this ecosystem.
The Galápagos Archipelago is located at the intersection of several major oceanographic features that produce diverse environmental conditions around the islands, and thus has the potential to serve as a natural laboratory for discerning the underlying environmental factors that structure marine microbial communities. Here we used quantitative metagenomics to characterize microbial communities in relation to archipelago marine habitats, and how those populations shift due to substantial environmental changes brought on by El Niño. Environmental conditions such as temperature, salinity, inorganic dissolved nutrients, and dissolved organic carbon (DOC) concentrations varied throughout the archipelago, revealing a diversity of potential microbial niches arising from upwelling, oligotrophic to eutrophic gradients, physical isolation, and potential island mass effects. The volumetric abundances of microbial community members shifted with these environmental changes and revealed several taxonomic indicators of different water masses. This included a transition from a Synechococcus dominated system in the west to an even mix of Synechococcus and Prochlorococcus in the east, mirroring the archipelago's mesotrophic to oligotrophic and productivity gradients. Several flavobacteria groups displayed characteristic habitat distributions, including enrichment of Polaribacter and Tenacibaculum clades in the relatively nutrient rich western waters, Leeuwenhoekiella spp. that were enriched in the more nutrient-deplete central and eastern sites, and the streamlined MS024-2A group found to be abundant across all sites. During the 2015/16 El Niño event, both environmental conditions and microbial community composition were substantially altered, primarily on the western side of the archipelago due to the reduction of upwelling from the Equatorial Undercurrent. When the upwelling resumed, concentrations of inorganic nutrients and DOC at the western surface sites were more typical of mesopelagic depths. Correspondingly, Synechococcus abundances decreased by an order of magnitude, while groups associated with deeper water masses were enriched, including streamlined roseobacters HTCC2255 and HIMB11,
The injection of supercritical CO2 into depleted oil reservoirs for long-term storage will influence the subsurface biogeochemistry with important implications for future carbon storage sites. In this study, we characterized produced water collected from CO2 enhanced oil recovery (CO2 EOR) well separators in the Niagaran Pinnacle Reef, an oil-producing region in the Michigan Basin that is a proposed target for future geological carbon storage. Our analysis investigated the geochemical and microbial community composition of produced water to understand the overall biogeochemistry in subsurface environments exposed to conditions expected during carbon storage in a depleted oil reservoir. The majority of sampled wells were characterized by high salinity and high total dissolved solids ranging from 122,000 to 416,000 mg/L. In addition, the sample wells contained supercritical concentrations of CO2 (scCO2) that appeared to drive low microbial community abundance among the sampled wells. We also observed minimal well-to-well communication, suggesting low permeability and reduced fluid migration within the reservoir system, which likely fostered isolated evolution of the reservoir microbiomes. The predominant microorganisms observed have been described in previous carbon storage systems, including Desulfotomaculum, Sulfurospirillum, Halanaerobium, Acetobacterium, and Pseudomonas. These taxa may participate in microbial processes such as sulfide and acid production, biofilm formation, and biomineralization. These metabolic processes can impact reservoir quality and stability and long-term carbon storage. Overall, these results suggest that these subsurface reservoirs select for stress-tolerant microbial communities that are adapted to high salinity and scCO2 exposure. This work contributes to a greater understanding of the site-specific microbiology in a reservoir targeted for long-term CO2 sequestration.
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