Enrichment of marine microbes to remove nitrogen of urea wastewater under salinity stress.

Salinity (NaCl) and urea concentration significantly affect the diversity, structural and physiological function of microbial communities in the biological treatment of wastewater. However, the responses of microbial in high salt and urea wastewater remain elusive. Here, we investigated microbial community function and assembly of four regions using gradient domestication experiment combined with 16S rRNA gene sequencing and statistical methods. The results showed that with the increase of salinity and urea concentration, the consortium Xiamen could still remove most urea, while the other three consortia could not. The alpha diversity of microbial community initially decreased and then increased, showing a recovery trend. After domestication, the consortium Xiamen exhibited high physiological activity and complex network structure, and the community assembly process changed from stochastic to deterministic during the domestication. Furthermore, the keystones with low abundance were associated with urea removal and important for maintain the complexity of the networks, while Arenibacter and Oceanimonas were found to be keystones in maintaining efficient urea removal in harsh environments. To sum up, environmental effects dominated by salinity and urea concentration stress drove the community assembly and species coexistence that underpinned the microbial differentiation pattern at a geographic scale. These results provided new sights for elucidate how microbial response to salinity and urea during wastewater treatment.

Insights into lignin bioconversion: lignin-derived compounds treatment of a novel marine fungus K-2.

BACKGROUND: The potential for the efficient conversion of lignocellulosic biomass has been extensively explored to produce a wide range of bioproducts. Many approaches have been sought for the deep conversion of lignin to generate products that are toxin-free and beneficial for processing into high-value-added components. RESULTS: This study reported a fungus isolated from the deep sea with strong synthesis of multiple lignocellulases, conversion of lignin and guaiacol (0.1%) by 71.6% and 86.1% within 9 days at 30 °C respectively, and outstanding environmental adaptability (20-50 °C and pH 3-8). Metabolic pathway profiling showed that this fungus utilized lignin to rapidly activate multiple ring-opening reactions including the 2,3- and 3,4-cleavage pathways, with the 2,3-cleavage pathway predominating after 5 days. Conversion of metabolic intermediates confirmed the superb potential of this strain for lignin treatment. Meanwhile, its shikimic acid pathway was metabolically active under lignin. CONCLUSION: This further expands the potential to produce valuable bioproducts during lignin treatment, especially under ambient conditions, which can significantly enhance high-value precursor compound production. © 2024 Society of Chemical Industry.

Moderate permeability enhanced microbial community turnover and copper extraction during bioleaching of low-grade copper ores.

Heap bioleaching is one of the most promising technologies for extracting valuable metals from low-grade ores. However, the effects of permeability of the heap on microbial community and bioleaching efficiency remain unclear. In this study, heap bioleaching systems with different permeability were constructed. Despite the high content of larger particles had better permeability (0.25 cm/s) and oxygen transfer efficiency (0.14), there was a 25 % decrease in copper extraction compared with the moderate permeability group (81.2 %), while low permeability (0.025 cm/s) could cut the extraction in half (48.5 %). The fine profiles of microbial communities based on relatively and absolutely quantitative technologies suggested that permeability significantly affected microbial diversity, biomass, and composition. Microbial community evenness was crucial to improving extraction than biomass. Additionally, Thermoplasmatales except for Acidiplasma and Ferroplasma played vital roles in bioleaching. This study highlighted the delicate trade-off of particle size-mediated permeability for intensifying bioleaching efficiency of low-grade copper ores.

Regulation of carbon metabolic fluxes to enhance lipid and succinate production in oleaginous fungus Mortierella alpina.

Mortierella alpina is popular for lipid production, but the low carbon conversion rate and lipid yield are major obstacles for its economic performance. Here, external addition of organic acids involved in tricarboxylic acid cycle was used to tune carbon flux and improve lipid production. Citrate was determined to be the best organic acid that can be used for enhancing lipid production. By the addition of citrate, the lipid titer and content were approximately 1.24 and 1.34 times higher, respectively. Meanwhile, citrate supplement also promoted the accumulation of succinate, an important value-added platform chemical. Owing to the improved lipid and succinate production through adding citrate, the carbon conversion rate of M. alpina reached up to 52.17%, much higher than that of the control group (14.11%). The addition of citrate could redistribute carbon flux by regulating the expression level of genes related to tricarboxylic acid cycle metabolism. More carbon fluxes flow to lipid and succinate synthesis, which greatly improved the carbon conversion efficiency of M. alpina. This study provides an effective and straightforward strategy with potential economic benefits to improve carbon conversion efficiency in M. alpina.

Redundancy and resilience of microbial community under aniline stress during wastewater treatment.

Aniline is one of the most toxic and widespread organic pollutants. Although biological treatment is cost-effective and generates minimal secondary pollution, microbial communities are significantly affected by high aniline concentrations, which result in low degradation efficiency. However, a comprehensive understanding of the microbial community response to aniline stress is lacking. Here, we performed a cyclic experiment with aniline concentrations (200, 600, 1200, 600, and 200 mg/L) to investigate the ability of microbial communities to recover their performance after exposure to high aniline concentrations. At aniline concentrations up to 600 mg/L, the bioreactor exhibited high aniline removal efficiency (almost 100 %). Comamonas, Zoogloea, and Delftia played crucial roles in removing aniline and microbial beta diversity changed. Additionally, alpha diversity and network complexity decreased with increasing aniline concentration, but these metrics recovered to their original levels when the aniline concentration was returned to 200 mg/L. Homogeneous and heterogeneous selection dominated microbial community assembly. Therefore, according to the observed variations in community structure and the recovery of keystones after aniline stress, microbial community redundancy and resilience are pivotal for ensuring system stability. Overall, this study provides valuable insights into the redundancy and resilience of microbial communities under aniline stress and establishes a scientific basis for managing and evaluating wastewater treatment plants.

Reductant-independent oxidative cleavage of cellulose by a novel marine fungal lytic polysaccharide monooxygenase.

Among the enzymes derived from fungus that act on polysaccharides, lytic polysaccharide monooxygenase (LPMOs) has emerged as a new member with complex reaction mechanisms and high efficiency in dealing with recalcitrant crystalline polysaccharides. This study reported the characteristics, structure, and biochemical properties of a novel LPMO from Talaromyces sedimenticola (namely MaLPMO9K) obtained from the Mariana Trench. MaLPMO9K was a multi-domain protein combined with main body and a carbohydrate-binding module. It was heterologously expressed in E. coli for analyzing peroxidase activity in reactions with the substrate 2,6-DMP, where H2O2 serves as a co-substrate. Optimal peroxidase activity for MaLPMO9K was observed at pH 8 and 25 °C, achieving the best Vmax value of 265.2 U·g-1. In addition, MaLPMO9K also demonstrated the ability to treat cellulose derivatives, and cellobiose substrates without the presence of reducing agents.

Bacterial abundant taxa exhibit stronger environmental adaption than rare taxa in the Arctic Ocean sediments.

Marine bacteria influence Earth's environmental dynamics in fundamental ways by controlling the biogeochemistry and productivity of the oceans. However, little is known about the survival strategies of their abundant and rare taxa, especially in polar marine environments. Here, bacterial environmental adaptation, community assembly processes, and co-occurrence patterns between abundant and rare taxa were compared in the Arctic Ocean sediments. Results indicated that the diversity of rare taxa is significantly higher than that of abundant taxa, whereas the distance-decay rate of rare taxa community similarity is over 1.5 times higher than that of abundant taxa. Furthermore, abundant taxa exhibited broader environmental breadth and stronger phylogenetic signals compared to rare taxa. Additionally, the community assembly processes of the abundant taxa were predominantly governed by 81% dispersal limitation, while rare taxa were primarily influenced by 48% heterogeneous selection. The co-occurrence network further revealed the abundant taxa formed a more complex network to enhance their environmental adaptability. This study revealed the differences in environmental responses and community assembly processes between bacterial abundant and rare taxa in polar ocean sediments, providing some valuable insights for understanding their environmental adaptation strategies in marine ecosystems.

Effects of Cd(II) on nitrogen removal by a heterotrophic nitrification aerobic denitrification bacterium Pseudomonas sp. XF-4.

Simultaneous heterotrophic nitrification and aerobic denitrification (SND) is gaining tremendous attention due to its high efficiency and low cost in water treatment. However, SND on an industrial scale is still immature since effects of coexisting pollutants, for example, heavy metals, on nitrogen removal remains largely unresolved. In this study, a HNAD bacterium (Pseudomonas sp. XF-4) was isolated. It could almost completely remove ammonium and nitrate at pH 5-9 and temperature 20 ℃-35 ℃ within 10 h, and also showed excellently simultaneous nitrification and denitrification efficiency under the coexistence of any two of inorganic nitrogen sources with no intermediate accumulation. XF-4 could rapidly grow again after ammonium vanish when nitrite or nitrate existed. There was no significant effects on nitrification and denitrification when Cd(II) was lower than 10 mg/L, and 95 % of Cd(II) was removed by XF-4. However, electron carrier and electron transport system activity was inhibited, especially at high concentration of Cd(II). Overall, this study reported a novel strain capable of simultaneous nitrification and denitrification coupled with Cd(II) removal efficiently. The results provided new insights into treatment of groundwater or wastewater contaminated by heavy metals and nitrogen.

Distinct strategies of the habitat generalists and specialists in the Arctic sediments: Assembly processes, co-occurrence patterns, and environmental implications.

Microorganisms could be classified as habitat generalists and specialists according to their niche breadth, uncovering their survival strategy is a crucial topic in ecology. Here, differences in environmental adaptation, community assemblies, co-occurrence patterns, and ecological functions between generalists and specialists were explored in the Arctic marine sediments. Compared to specialists, generalists showed lower alpha diversity but stronger environmental adaption, and dispersal limitation contributed more to the community assembly of specialists (74 %) than generalists (46 %). Furthermore, the neutral theory model demonstrated that generalists (m = 0.20) had a higher immigration rate than specialists (m = 0.02), but specialists exhibited more complex co-occurrence patterns than generalists. Our results also found that generalists may play more important roles in C, N, S metabolism but are weaker in carbon fixation and xenobiotic biodegradation and metabolism. This study would broaden our understanding of bacterial generalists' and specialists' survival strategies, and further reveal their ecological functions in marine sediments.

Integration of lactic acid biorefinery with treatment of red mud from alumina refinery: win-win paradigm for waste valorization.

The cost of detoxification and neutralization poses certain challenges to the development of an economically viable lactic acid biorefinery with lignocellulosic biomass as feedstock. Herein, red mud, an alkaline waste, was explored as both a detoxifying agent and a neutralizer. Red mud treatment of lignocellulosic hydrolysate effectively removed the inhibitors generated in dilute acid pretreatment, improving the lactic acid productivity from 1.0 g/L·h-1 to 1.9 g/L·h-1 in later fermentation. In addition, red mud could replace CaCO3 as a neutralizer in lactic acid fermentation, which in turn enabled simultaneous bioleaching of valuable metals (Sc, Y, Nd, and Al) from red mud. The neutralization of alkali in red mud by acids retained in lignocellulosic hydrolysate and lactic acid produced from fermentation led to effective dealkalization, rendering a maximum alkali removal efficiency of 92.2 %. Overall, this study offered a win-win strategy for the valorization of both lignocellulosic biomass and red mud.

Treating waste with waste: Lignin acting as both an effective bactericide and passivator to prevent acid mine drainage formation at the source.

Acid mine drainage (AMD) induced by pyrite oxidation is a notorious and serious environmental problem, but the management of AMD in an economical and environmentally friendly way remains challenging. Here, lignin, a natural polymer and abundant waste, was employed as both a bactericide and passivator to prevent AMD formation. The addition of lignin to a mimic AMD formation system inoculated with Acidithiobacillus ferrooxidans at a lignin-to-pyrite weight ratio of 2.5: 10 reduced the combined abiotic and biotic oxidation of pyrite by 68.4 % (based on released SO42-). Morphological characterization of Acidithiobacillus ferrooxidans revealed that lignin could act on the cell surface and impair the cell integrity, disrupting its normal growth and preventing biotic oxidation of pyrite accordingly. Moreover, lignin can be used alone as a passivator to form a coating on the pyrite surface, reducing abiotic oxidation by 71.7 % (based on released SO42-). Through multiple technique analysis, it was proposed that the functional groups on lignin may coordinate with iron ions on pyrite, promoting its deposition on the surface. In addition, the inherent antioxidant activity of lignin may also be actively involved in the abatement of pyrite oxidation via the reduction of iron. Overall, this study offered a "treating waste with waste" strategy for preventing AMD formation at the source and opened a new avenue for the management of AMD.

Discrepant assembly processes of prokaryotic communities between the abyssal and hadal sediments in Yap Trench.

Abyssal and hadal sediments represent two of the most type ecosystems on Earth and have the potential interactions with geochemistry. However, little is known about the prokaryotic community assembly and the response of prokaryotic communities to metal(loid)s in trench sediments due to the lack of adequate and appropriate samples. In this study, a systematic investigation combined the assembly mechanisms and co-occurrence patterns of prokaryotic communities between the hadal and abyssal sediments across the Yap Trench. The results revealed that the hadal prokaryotes had less species diversity, but more abundant function than the abyssal prokaryotes. The prokaryotic communities in the abyssal sediments had more core taxa than the hadal sediments. Twenty-one biomarkers mostly affiliated with Nitrosopumilaceae were detected using Random-Forests machine learning algorithm. Furthermore, stochasticity was dominant in the prokaryotic community assembly processes of the Yap Trench sediments. Meanwhile, homogeneous selection (32.6%-52.9%) belonging to deterministic processes governed the prokaryotic community assembly in hadal sediments with increasing of sediment depth. In addition to total nitrogen and total organic carbon, more metal(loid)s were significantly correlated with the prokaryotic community in the hadal sediments than that in the abyssal sediments. The hadal prokaryotic communities was most positively related to bismuth (r = 0.31, p < 0.01), followed by calcium, chromium, cerium, potassium, plumbum, scandium, titanium, and vanadium. Finally, co-occurrence networks revealed two potential dominant prokaryotic modules in Yap Trench sediments covaried across oceanographic zonation. By contrast, the hadal network had relatively more complexity, more bacterial taxa, and more associations among prokaryotic taxa, relative to the abyssal network. This study reveals potentially metal variables and community assembly mechanisms of the prokaryotic community in abyssal and hadal sediments and provides a better understanding on the prokaryotic diversity and ecology in trench sediment ecosystems.

Depth significantly affects plastisphere microbial evenness, assembly and co-occurrence pattern but not richness and composition.

Microplastics have become one of the hot concerns of global marine pollution. In recent years, diversity and abiotic influence factors of plastisphere microbial communities were well documented, but our knowledge of their assembly mechanisms and co-occurrence patterns remains unclear, especially the effects of depth on them. Here, we collected microorganisms on microplastics to investigate how ocean depth affects on microbial diversity, community composition, assembly processes and co-occurrence patterns. Our results indicated that there were similar microbial richness and community compositions but microbial evenness and unique microbes were obviously different in different ocean layers. Our findings also demonstrated that deterministic processes played dominant roles in the assembly of the mesopelagic plastisphere microbial communities, while the bathypelagic microbial community assembly was mainly shaped by stochastic processes. In addition, the co-occurrence networks suggested that the relationships between microorganisms in the mesopelagic layer were more complex and stable than those in the bathypelagic layer. Simultaneously, we also found that Proteobacteria and Actinobacteriota were the most abundant keystones which played important roles in microbial co-occurrence networks at both layers. This study enhanced our understanding of microbial diversity, assembly mechanism, and co-occurrence pattern on plastisphere surfaces, and provided useful insights into microorganisms capable of degrading plastics and microbial remediation.

Depth-Dependent Distribution of Prokaryotes in Sediments of the Manganese Crust on Nazimov Guyots of the Magellan Seamounts.

Deep ocean polymetallic nodules, rich in cobalt, nickel, and titanium which are commonly used in high-technology and biotechnology applications, are being eyed for green energy transition through deep-sea mining operations. Prokaryotic communities underneath polymetallic nodules could participate in deep-sea biogeochemical cycling, however, are not fully described. To address this gap, we collected sediment cores from Nazimov guyots, where polymetallic nodules exist, to explore the diversity and vertical distribution of prokaryotic communities. Our 16S rRNA amplicon sequencing data, quantitative PCR results, and phylogenetic beta diversity indices showed that prokaryotic diversity in the surficial layers (0-8 cm) was > 4-fold higher compared to deeper horizons (8-26 cm), while heterotrophs dominated in all sediment horizons. Proteobacteria was the most abundant taxon (32-82%) across all sediment depths, followed by Thaumarchaeota (4-37%), Firmicutes (2-18%), and Planctomycetes (1-6%). Depth was the key factor controlling prokaryotic distribution, while heavy metals (e.g., iron, copper, nickel, cobalt, zinc) can also influence significantly the downcore distribution of prokaryotic communities. Analyses of phylogenetic diversity showed that deterministic processes governing prokaryotic assembly in surficial layers, contrasting with stochastic influences in deep layers. This was further supported from the detection of a more complex prokaryotic co-occurrence network in the surficial layer which suggested more diverse prokaryotic communities existed in the surface vs. deeper sediments. This study expands current knowledge on the vertical distribution of benthic prokaryotic diversity in deep sea settings underneath polymetallic nodules, and the results reported might set a baseline for future mining decisions.

Regulation on Pathway Metabolic Fluxes to Enhance Colanic Acid Production in Escherichia coli.

Colanic acid (CA) is a natural polysaccharide macromolecule with rich and unique biological properties and is a promising candidate for use in food and cosmetics. To date, the efficient biosynthesis of CA and the influence of product accumulation on the strains used have yet to be precisely investigated. Herein, bottlenecks in the CA metabolic pathway were untangled by finely regulating the expression of manA, cpsG, fcl, and rcsA. Engineered strains produced CA at >1 g/L in shake flasks without dependence on cold temperatures, and it was verified in a 1 L bioreactor with a titer up to 18.64 g/L within 24 h. The accumulation of CA caused a decrease in the saturated fatty acid content (represented by C16:0 and C18:0) in the cell membrane. This study demonstrated pathway engineering for efficient CA production in cell factories and provided insights into the barriers and solutions faced in the biosynthesis of natural products.

Enhanced Enzymatic Performance of ß-Mannanase Immobilized on Calcium Alginate Beads for the Generation of Mannan Oligosaccharides.

Mannan oligosaccharides (MOSs) are excellent prebiotics that are usually obtained via the enzymatic hydrolysis of mannan. In order to reduce the cost of preparing MOSs, immobilized enzymes that demonstrate good performance, require simple preparation, and are safe, inexpensive, and reusable must be developed urgently. In this study, ß-mannanase was immobilized on calcium alginate (CaAlg). Under the optimal conditions of 320 U enzyme addition, 1.6% sodium alginate, 2% CaCl2, and 1 h of immobilization time, the immobilization yield reached 68.3%. The optimum temperature and pH for the immobilized ß-mannanase (Man-CaAlg) were 75 °C and 6.0, respectively. The Man-CaAlg exhibited better thermal stability, a high degree of pH stability, and less substrate affinity than free ß-mannanase. The Man-CaAlg could be reused eight times and retained 70.34% of its activity; additionally, the Man-CaAlg showed 58.17% activity after 30 days of storage. A total of 7.94 mg/mL of MOSs, with 4.94 mg/mL of mannobiose and 3.00 mg/mL of mannotriose, were generated in the oligosaccharide production assay. It is believed that this convenient and safe strategy has great potential in the important field of the use of immobilized ß-mannanase for the production of mannan oligosaccharides.

Distinct assembly processes and environmental adaptation of abundant and rare archaea in Arctic marine sediments.

Revealing the ecological processes and environmental adaptation of abundant and rare archaea is a central, but poorly understood, topic in ecology. Here, abundant and rare archaeal diversity, community assembly processes and co-occurrence patterns were comparatively analyzed in Arctic marine sediments. Our findings revealed that the rare taxa exhibited significantly higher diversity compared to the abundant taxa. Additionally, the abundant taxa displayed stronger environmental adaptation than the rare taxa. The co-occurrence network analysis demonstrated that the rare taxa developed more interspecies interactions and modules in response to environmental disturbance. Furthermore, the community assembly of abundant and rare taxa in sediments was primarily controlled by stochastic and deterministic processes, respectively. These findings provide valuable insights into the archaeal community assembly processes and significantly contribute to a deeper understanding of the environmental adaptability of abundant and rare taxa in Arctic marine sediments.

Kinetic process of the biosorption of Cu(II), Ni(II) and Cr(VI) by waste Pichia pastoris cells.

Waste biomass of Pichia pastoris (P.pastoris) cells from the fermentation industry is an environmentally friendly biosorption material. The present study aimed to explore the biosorption behaviour of waste P.pastoris cells for Cu(II), Ni(II) and Cr(VI) in aqueous solution conditions. The results showed that the adsorption kinetics of three kinds of metals were well-fitted with lineared Elovich, pseudo-second-order kinetics models, non-linear kinetics and adsorption isotherms. The effective biosorption rates for Cu(II), Ni(II) and Cr(VI) removal were 71.3%, 59.7% and 16.25% respectively. The maximum Cu(II) adsorption capacity of waste P.pastoris was 40 mg/g at pH = 4 and 225 mg/L of solute concentration for 0.4 g biomass, better than that of the living yeasts. The pattern of Fourier transform infrared (FTIR) indicated that functional groups such as -NH, -OH, Si-O, P-O-C were involved in Cu(II) adsorption process. The analysis of SEM-EDS, XRD and TEM-EDS can be concluded that Cu(II) occupied Ca(II) binding sites by ion exchange mechanism to remove flocculation, and Cu(II) adsorbed onto the diatomite containing in the industrial waste P.pastoris. Thus the adsorption mechanism of the industrial waste P.pastoris was proposed taking Cu(II) as the example. And consecutive biosorption/desorption cycles were used for the evaluation of the regeneration efficiency, suggesting the good regeneration and reusability of waste P.pastoris.

Enhancing lactose recognition of a key enzyme in 2'-fucosyllactose synthesis: α-1,2-fucosyltransferase.

BACKGROUND: 2'-Fucosyllactose, a representative oligosaccharide in human milk, is an emerging and promising food and pharmaceutical ingredient due to its powerful health benefits, such as participating in immune regulation, regulation of intestinal flora, etc. To enable economically viable production of 2'-fucosyllactose, different biosynthesis strategies using precursors and pathway enzymes have been developed. The α-1,2-fucosyltransferases are an essential part involved in these strategies, but their strict substrate selectivity and unsatisfactory substrate tolerance are one of the key roadblocks limiting biosynthesis. RESULTS: To tackle this issue, a semi-rational manipulation combining computer-aided designing and screening with biochemical experiments were adopted. The mutant had a 100-fold increase in catalytic efficiency compared to the wild-type. The highest 2'-fucosyllactose yield was up to 0.65 mol mol-1 lactose with a productivity of 2.56 g mL-1  h-1 performed by enzymatic catalysis in vitro. Further analysis revealed that the interactions between the mutant and substrates were reduced. The crucial contributions of wild-type and mutant to substrate recognition ability were closely related to their distinct phylotypes in terms of amino acid preference. CONCLUSION: It is envisioned that the engineered α-1,2-fucosyltransferase could be harnessed to relieve constraints imposed on the bioproduction of 2'-fucosyllactose and lay a theoretical foundation for elucidating the substrate recognition mechanisms of fucosyltransferases. © 2022 Society of Chemical Industry.

Temperature mediated the balance between stochastic and deterministic processes and reoccurrence of microbial community during treating aniline wastewater.

Seasonal temperature changes significantly affect microbial community diversity, composition, and performance in wastewater treatment plants. However, the community assembly mechanisms under seasonal temperature variations remain unclear. Here, we carried out temperature cycling experiments (30 °C, 35 °C, 37 °C, 40 °C, 42 °C, 45 °C, 40 °C, and 30 °C) to investigate how temperature impacts microbial performance and co-occurrence network and how assembly processes determine the structure and function of microbial communities during treating aniline wastewater. During the 195-day operation, the system achieved an efficient and stable aniline removal of 99%. Interestingly, α-diversity and network complexity were negatively correlated with temperature but could be recovered when the temperature was returned to 30 °C. The results showed that functional redundancy was probably responsible for the excellent microbial performance during the whole process. Null model analyses presented that deterministic process dominated the community when the temperature was 30 °C, and stochasticity dominated the assembly process when the temperature was over 30 °C. Overall, the balance between stochastic and deterministic processes in the treatment of aniline wastewater mediated the reoccurrence of microbial community and co-occurrence network at different temperatures. This study provides new insights into microbial community reoccurrence under seasonal temperature changes and a theoretical basis for regulating microbial communities in wastewater treatment plants.