Efficient Enhancement of Extracellular Electron Transfer in Shewanella oneidensis MR-1 via CRISPR-Mediated Transposase Technology.

Electroactive bacteria, exemplified by Shewanella oneidensis MR-1, have garnered significant attention due to their unique extracellular electron-transfer (EET) capabilities, which are crucial for energy recovery and pollutant conversion. However, the practical application of MR-1 is constrained by its EET efficiency, a key limiting factor, due to the complexity of research methodologies and the challenges associated with the practical use of gene editing tools. To address this challenge, a novel gene integration system, INTEGRATE, was developed, utilizing CRISPR-mediated transposase technologies for precise genomic insertion within the S. oneidensis MR-1 genome. This system facilitated the insertion of extensive gene segments at different sites of the Shewanella genome with an efficiency approaching 100%. The inserted cargo genes could be kept stable on the genome after continuous cultivation. The enhancement of the organism's EET efficiency was realized through two primary strategies: the integration of the phenazine-1-carboxylic acid synthesis gene cluster to augment EET efficiency and the targeted disruption of the SO3350 gene to promote anodic biofilm development. Collectively, our findings highlight the potential of utilizing the INTEGRATE system for strategic genomic alterations, presenting a synergistic approach to augment the functionality of electroactive bacteria within bioelectrochemical systems.

Transcriptomic Insights into Metabolism-Dependent Biosynthesis of Bacterial Nanocellulose.

Bacterial nanocellulose (BNC) is an attractive green-synthesized biomaterial for biomedical applications and various other applications. However, effective engineering of BNC production has been limited by our poor knowledge of the related metabolic processes. In contrast to the traditional perception that genome critically determines biosynthesis behaviors, here we discover that the glucose metabolism could also drastically affect the BNC synthesis in Gluconacetobacter hansenii. The transcriptomic profiles of two model BNC-producing strains, G. hansenii ATCC 53582 and ATCC 23769, which have highly similar genomes but drastically different BNC yields, were compared. The results show that their BNC synthesis capacities were highly related to metabolic activities such as ATP synthesis, ion transport protein assembly, and carbohydrate metabolic processes, confirming an important role of metabolism-related transcriptomes in governing the BNC yield. Our findings provide insights into the microbial biosynthesis behaviors from a transcriptome perspective, potentially guiding cellular engineering for biomaterial synthesis.

Nonsterilized Fermentation of Crude Glycerol for Polyhydroxybutyrate Production by Metabolically Engineered Vibrio natriegens.

Polyhydroxybutyrate (PHB) is an attractive biodegradable polymer that can be produced through the microbial fermentation of organic wastes or wastewater. However, its mass production has been restricted by the poor utilization of organic wastes due to the presence of inhibitory substances, slow microbial growth, and high energy input required for feedstock sterilization. Here, Vibrio natriegens, a fast-growing bacterium with a broad substrate spectrum and high tolerance to salt and toxic substances, was genetically engineered to enable efficient PHB production from nonsterilized fermentation of organic wastes. The key genes encoding the PHB biosynthesis pathway of V. natriegens were identified through base editing and overexpressed. The metabolically engineered strain showed 166-fold higher PHB content (34.95 wt %) than the wide type when using glycerol as a substrate. Enhanced PHB production was also achieved when other sugars were used as feedstock. Importantly, it outperformed the engineered Escherichia coli MG1655 in PHB productivity (0.053 g/L/h) and tolerance to toxic substances in crude glycerol, without obvious activity decline under nonsterilized fermentation conditions. Our work demonstrates the great potential of engineered V. natriegens for low-cost PHB bioproduction and lays a foundation for exploiting this strain as a next-generation model chassis microorganism in synthetic biology.

Genome Sequence of a Heavy Metal-Detoxifying Actinomycete, Microbacterium proteolyticum ustc.

A complete genome is presented for Microbacterium proteolyticum ustc, a member of the Gram-positive order Micrococcales of the phylum Actinomycetota that is resistant to high concentrations of heavy metals and participates in metal detoxification. The genome consists of one plasmid and one chromosome.

AQDS Activates Extracellular Synergistic Biodetoxification of Copper and Selenite via Altering the Coordination Environment of Outer-Membrane Proteins.

The biotransformation of heavy metals in the environment is usually affected by co-existing pollutants like selenium (Se), which may lower the ecotoxicity of heavy metals, but the underlying mechanisms remain unclear. Here, we shed light on the pathways of copper (Cu2+) and selenite (SeO32-) synergistic biodetoxification by Shewanella oneidensis MR-1 and illustrate how such processes are affected by anthraquinone-2,6-disulfonate (AQDS), an analogue of humic substances. We observed the formation of copper selenide nanoparticles (Cu2-xSe) from synergistic detoxification of Cu2+ and SeO32- in the periplasm. Interestingly, adding AQDS triggered a fundamental transition from periplasmic to extracellular reaction, enabling 14.7-fold faster Cu2+ biodetoxification (via mediated electron transfer) and 11.4-fold faster SeO32- detoxification (via direct electron transfer). This is mainly attributed to the slightly raised redox potential of the heme center of AQDS-coordinated outer-membrane proteins that accelerates electron efflux from the cells. Our work offers a fundamental understanding of the synergistic detoxification of heavy metals and Se in a complicated environmental matrix and unveils an unexpected role of AQDS beyond electron mediation, which may guide the development of more efficient environmental remediation and resource recovery biotechnologies.

Anaerobic self-assembly of a regenerable bacteria-quantum dot hybrid for solar hydrogen production.

Inorganic-biological hybrid systems (bio-hybrids), comprising fermentative bacteria and inorganic semiconductor photosensitizers for synergistic utilization of solar energy and organic wastes, offer opportunities for sustainable fuel biosynthesis, but the low quantum efficiency, photosensitizer biotoxicity and inability for self-regeneration are remaining hurdles to practical application. Here, we unveil a previously neglected role of oxygen in suppressing the biosynthesis of cadmium selenide quantum dots (CdSe QDs) and the metabolic activities of Escherichia coli, and accordingly propose a simple oxygen-regulation strategy to enable the self-assembly of bacterial-QD hybrids for efficient solar hydrogen production. Shifting from aerobic to anaerobic biosynthesis significantly lowered the intracellular reactive oxygen species level and increased NADPH and thiol-protein production, enabling a two-order-of-magnitude higher bio-QD synthesis rate and resulting in CdSe-rich products. Bacteria with abundant biocompatible intracellular bio-QDs naturally formed a highly active and self-regenerable bio-hybrid and achieved a quantum efficiency of 28.7% for hydrogen production under visible light, outperforming all the existing bio-hybrids. It also exhibited high stability during cyclic operation and robust performance for treating real wastewater under simulated sunlight. Our work provides valuable new insights into the metallic nanomaterial biosynthesis process to guide the design of self-assembled bio-hybrids towards sustainable energy and environmental applications.

Reductions in crop yields across China from elevated ozone.

Exposure of crops to high concentrations of ozone can cause substantial reductions in yield that pose a serious threat to global food security. Here we provide comprehensive estimates of yield losses for key crops across China between 2014 and 2017 attributed to ozone using a number of new approaches. We use an air quality model at 5 km resolution and crop-specific dose-response functions developed for both concentration- and flux-based metrics. We bias correct modelled ozone concentrations and metrics using observations from more than 1000 locations. We find that on a 4-year average basis, production losses of key crops are 34-91 million metric tonnes (Mt/yr), dependent on the approach used, with highest losses in Henan province. At a national level, loss of winter wheat production derived using a China-specific dose-response function increased by 82% from 2014 to 2017, with large interannual variations in the North China Plain and in eastern China. Winter wheat losses estimated using flux-based functions, which require robust simulation of stomatal conductance and underlying vegetation physiology, are significantly lower, at 30 Mt/yr. We show that the definition of the growing season may have a greater impact on estimated losses than small biases in ozone surface concentrations. Although uncertainties remain, our findings demonstrate that increasing ozone concentrations have substantial adverse impacts on crop yields and threaten food security in China. It is important to control ozone concentrations to mitigate these negative impacts.

Mechanical Insights into Thiol-Mediated Synergetic Biotransformation of Cadmium and Selenium in Nematodes.

Cadmium ion (Cd2+) is a common environmental pollutant with high biotoxicity. Interestingly, the Cd2+ biotoxicity can be alleviated by the coexisting selenite (SeO32-), which induces the formation of cadmium selenide-rich nanoparticles (CdSe NPs) under the function of thiol-capping peptides. However, the detailed biochemical mechanisms by which Cd and Se are synergistically transformed into CdSe NPs in living organisms remain unclear so far. Here, we shed light on the molecular basis of such biotransformation processes in Caenorhabditis elegans by focusing on the roles of several key thiol-capping peptides. By monitoring the compositional and structural changes of the Cd and Se species and the genetic-level responses of nematodes, we revealed the specific roles of glutathione (GSH) and phytochelatins (PCs) in mediating the CdSe NP formation. With the aid of in vitro bioassembly assay and density functional theory calculations, the detailed Cd-Se interaction pathways were further deciphered: the ingested Cd binds predominantly to GSH and PCs in sequence, then further interacts with selenocysteine to form tetrahedral-structured PC2-Cd2-Sec2 complex, and ultimately grows into CdSe NPs. This work provides molecular-level insights into the Cd-Se interaction in C. elegans and lays a basis for controlling the ecological and health risks of heavy metals in polluted environment.

Distribution and evolution of H1N1 influenza A viruses with adamantanes-resistant mutations worldwide from 1918 to 2019.

H1N1 influenza is a kind of acute respiratory infectious disease that has a high socioeconomic and medical burden each year around the world. In the past decades, H1N1 influenza viruses have exhibited high resistance to adamantanes, which has become a serious issue. To understand the up-to-date distribution and evolution of H1N1 influenza viruses with adamantanes-resistant mutations, we conducted a deep analysis of 15875 M2 protein and 8351 MP nucleotides sequences. Results of the distribution analyses showed that 77.32% of H1N1 influenza viruses harbored-resistance mutations of which 73.52% were S31N, And the mutant variants mainly appeared in North America and Europe and H1N1 influenza viruses with S31N mutation became the circulating strains since 2009 all over the world. In addition, 80.65% of human H1N1 influenza viruses and 74.61% of swine H1N1 influenza viruses exhibited adamantanes resistance, while the frequency was only 1.86% in avian H1N1 influenza viruses. Studies from evolutionary analyses indicated that the avian-origin swine H1N1 influenza viruses replaced the classical human H1N1 influenza viruses and became the circulating strains after 2009; The interspecies transmission among avian, swine, and human strains over the past 20 years contributed to the 2009 swine influenza pandemic. Results of our study clearly clarify the historical drug resistance level of H1N1 influenza viruses around the world and demonstrated the evolution of adamantanes-resistant mutations in H1N1 influenza viruses. Our findings emphasize the necessity for monitoring the adamantanes susceptibility of H1N1 influenza viruses and draw attention to analyses of the evolution of drug-resistant H1N1 influenza variants.

Health impacts of long-term ozone exposure in China over 2013-2017.

Increasing ozone concentrations are becoming a severe problem for air pollution in China and have an adverse impact on human health. Here we evaluate premature deaths attributable to long-term exposure to ambient ozone in China between 2013 and 2017 with an air quality model at 5 km resolution and the latest estimates of the relative risk to health. We use a modified inverse distance weighting method to bias-correct the key model-simulated ozone metrics. We find that on a 5-year average basis there are 186,000 (95% Confidence Interval: 129,000-237,000) respiratory deaths and 125,000 (42,000-204,000) cardiovascular deaths attributable to ozone exposure. Sichuan exhibits the largest per capita respiratory mortality (0.31‰) among all provinces. We find that there are 73,000 (51,000-93,000) premature respiratory deaths in urban areas, accounting for 39% of total deaths. Between 2013 and 2017 the population-weighted annual average maximum daily 8-h average ozone (AMDA8) and premature respiratory deaths increased by 14% and 31%, respectively, at a national level. Changes in precursor emissions explain most of these increases, with differences in meteorology accounting for 21% and 16% respectively. Interannual variations in population-weighted ozone and premature respiratory deaths at a provincial level are much larger than those at a national level, particularly in northern, central and eastern China. These findings emphasize that ozone should be an important focus of future air quality policies in China, and tighter controls of precursor emissions are urgently needed.

Using the Google Earth Engine to estimate a 10 m resolution monthly inventory of soil fugitive dust emissions in Beijing, China.

Soil fugitive dust (SFD) is an important contributor to ambient particulate matter (PM), but most current SFD emission inventories are updated slowly or have low resolution. In areas where vegetation coverage and climatic conditions undergo significant seasonal changes, the classic wind erosion equation (WEQ) tends to underestimate SFD emissions, increasing the need for higher spatiotemporal data resolution. Continuous acquisition of precise bare soil maps is the key barrier to compiling monthly high-resolution SFD emission inventories. In this study, we proposed taking advantage of the massive Landsat and Sentinel-2 imagery data sets stored in the Google Earth Engine (GEE) cloud platform to enable the rapid production of bare soil maps with spatial resolutions of up to 10 m. The resulting improved spatiotemporal resolution of wind erosion parameters allowed us to estimate SFD emissions in Beijing as being ~5-7 times the level calculated by the WEQ. Spring and winter accounted for >85% of SFD emissions, while April was the dustiest month with SFD emissions of PM10 exceeding 11,000 t. Our results highlighted the role of SFD in air pollution during winter and spring in northern China, and suggested that GEE should be further used for image acquisition, data processing, and compilation of gridded SFD inventories. These inventories can help identify the location and intensity of SFD sources while providing supporting information for local authorities working to develop targeted mitigation measures.

Numerical study of air pollution over a typical basin topography: Source appointment of fine particulate matter during one severe haze in the megacity Xi'an.

Many cities are located in lands with typical basin topographies, which are not conducive to the spread of air pollutants. In the winter of 2016/2017, a severe haze happened in Xi'an, the main city in the Guanzhong Basin in central China. When the peak daily concentration of fine particulate matter (PM2.5) reaches 499 µg/m3, the source of the atmospheric pollution needs to be found urgently in order to take countermeasures. The comprehensive air quality model with extensions, coupled with the tracer tagging particulate source apportionment technology (PSAT) module, and an improved emission inventory, higher grid resolution, and bigger inner domain area, have been applied to quantify the contributions of local and regional emissions to the PM2.5 pollutions. The model performed well in time period considered in this study. The correlation of the simulated daily PM2.5 concentration data reaches 0.82, and the fraction of predictions within a factor of two of observations approaches 84%. With the PSAT module, the PM2.5 contributions from local and regional sources to the urban centre and rural areas during the severe winter haze event are analysed in detail. The PM2.5 concentrations in the urban centre in Xi'an is mainly originating from local emissions (60%), and Xianyang City is the largest contributor among the surrounding source regions (11.6%), while the transportation sector outside the Shaanxi Province (5.1%) also contributes significantly. Comparatively, the rural areas have lower local contributions and higher transport contributions. In particular, in the northern rural area Yanliang, the contribution from surrounding source regions approaches 82%. The results of this study suggest that to improve the air quality in a typical basin city, a regional-scale coordinated emissions control should be used, focusing on the emissions from both local and surrounding areas.

Improved Inversion of Monthly Ammonia Emissions in China Based on the Chinese Ammonia Monitoring Network and Ensemble Kalman Filter.

Ammonia (NH3) emission inventories are an essential input in chemical transport models and are helpful for policy-makers to refine mitigation strategies. However, current estimates of Chinese NH3 emissions still have large uncertainties. In this study, an improved inversion estimation of NH3 emissions in China has been made using an ensemble Kalman filter and the Nested Air Quality Prediction Modeling System. By first assimilating the surface NH3 observations from the Ammonia Monitoring Network in China at a high resolution of 15 km, our inversion results have provided new insights into the spatial and temporal patterns of Chinese NH3 emissions. More enhanced NH3 emission hotspots, likely associated with industrial or agricultural sources, were captured in northwest China, where the a posteriori NH3 emissions were more than twice the a priori emissions. Monthly variations of NH3 emissions were optimized in different regions of China and exhibited a more distinct seasonality, with the emissions in summer being twice those in winter. The inversion results were well-validated by several independent datasets that traced gaseous NH3 and related atmospheric processes. These findings highlighted that the improved inversion estimation can be used to advance our understanding of NH3 emissions in China and their environmental impacts.

Developed and developing world responsibilities for historical climate change and CO2 mitigation.

At the United Nations Framework Convention on Climate Change Conference in Cancun, in November 2010, the Heads of State reached an agreement on the aim of limiting the global temperature rise to 2 °C relative to preindustrial levels. They recognized that long-term future warming is primarily constrained by cumulative anthropogenic greenhouse gas emissions, that deep cuts in global emissions are required, and that action based on equity must be taken to meet this objective. However, negotiations on emission reduction among countries are increasingly fraught with difficulty, partly because of arguments about the responsibility for the ongoing temperature rise. Simulations with two earth-system models (NCAR/CESM and BNU-ESM) demonstrate that developed countries had contributed about 60-80%, developing countries about 20-40%, to the global temperature rise, upper ocean warming, and sea-ice reduction by 2005. Enacting pledges made at Cancun with continuation to 2100 leads to a reduction in global temperature rise relative to business as usual with a 1/3-2/3 (CESM 33-67%, BNU-ESM 35-65%) contribution from developed and developing countries, respectively. To prevent a temperature rise by 2 °C or more in 2100, it is necessary to fill the gap with more ambitious mitigation efforts.