Extracellular Enzyme Activities Research Articles

In Situ Galacto-Oligosaccharides Synthesis in Whey Powder Fortified Milk by a Modified β-Galactosidase and Its Effect on the Techno-Functional Characteristics of Yogurt.

In situ galacto-oligosaccharide (GOS) synthesis in milk using β-galactosidases is an effective method for developing prebiotic dairy products. However, the low lactose concentration in milk (∼4.6%, w/w) reduces the GOS yield. In this study, a modified β-galactosidase from Bacillus circulans (mBgaD-D) with enhanced transglycosylation activity at low lactose concentration was developed through directed evolution and saturation mutagenesis. The GOS yield by mBgaD-D increased from 22.8% (wild type) to 30.8% in 50 g/L lactose (phosphate buffer). Pmgut was a strong sorbitol-inducible promoter from Bacillus subtilis. The expression of mBgaD-D in B. subtilis, coupled with the Pmgut promoter, resulted in a 6.4-fold increase (compared to the P43 promoter) in extracellular enzyme activity. Additionally, adding whey powder to boost the initial lactose concentration further improved the GOS yield, which reached 43% under the optimized conditions. Combining mBgaD-D and whey powder enhanced milk sweetness, producing no sugar-added, GOS-enriched yogurt (GOSY). The GOS content in GOSY was 4.1/100 g, providing an appropriate level of sweetness and yielding a yogurt that is elastic as well as firm. GOSY also increased the population of Bifidobacterium spp. during a 24 h in vitro fecal fermentation. Thus, fortifying yogurt with mBgaD-D and whey powder can enhance its technological properties and health benefits.

Microbial Life in Playa-Lake Sediments: Adapted Structure, Plastic Function to Extreme Water Activity Variations

Saline shallow lakes in arid and semi-arid regions frequently undergo drying episodes, leading to significant variations in salinity and water availability. Research on the impacts of salinity and drought on the structure and function of biofilms in hypersaline shallow lakes is limited. This study aimed to understand the potential changes of biofilms in playa-lake sediments during the drying process. Sediments were sampled at different depths (surface, subsurface) and hydrological periods (wet, retraction, and dry), which included a decrease in water activity (aw, the availability of water for microbial use) from 0.99 to 0.72. aw reduction caused a greater effect on functional variables compared to structural variables, indicating the high resistance of the studied biofilms to changes in salinity and water availability. Respiration and hydrolytic extracellular enzyme activities exhibited higher values under high aw, while phenol oxidase activity and prokaryote biomass increased at lower aw. This shift occurred at both depths but was more pronounced at the surface, possibly due to the more extreme conditions (up to 0.7 aw). The increased levels of extracellular polymeric substances and carotenoids developed at low aw may help protect microorganisms in high salinity and drought environments. However, these harsh conditions may interfere with the activity of hydrolytic enzymes and their producers, while promoting the growth of resistant prokaryotes and their capacity to obtain C and N sources from recalcitrant compounds. The resilience of biofilms in hypersaline lakes under extreme conditions is given by their resistant biochemichal structure and the adaptability of their microbial functioning.

Spatio-temporal distribution and biotechnological potential of culturable yeasts in the intertidal sediments and seawater of Aoshan Bay, China.

Marine yeasts play a crucial role in marine microbial ecology, facilitating the biogeochemical cycling of carbon and nitrogen in marine ecosystems, while also serving as important reservoirs of bioactive compounds with extensive applications in pharmaceuticals, agriculture, and various industries. Intertidal flats, characterized by their complex ecological dynamics, are postulated to harbor a wealth of yeast resources. This study employed a culture-dependent approach to assess the diversity, spatio-temporal distribution, and biotechnological potential of yeast communities residing within the intertidal sediments and seawater of Aoshan Bay. A total of 392 yeast strains were identified from 20 distinct genera, encompassing 43 recognized species and four candidate novel species. Notably, 17 of these species were identified as novel occurrences in marine environments, underscoring the rich yeast biodiversity of the Aoshan Bay ecosystem, with Candida emerging as the dominant genus in both sedimentary and aqueous habitats. Yeast community composition exhibited significant spatial and temporal variation, with peak diversity and abundance observed in autumn, the subtidal zone, and the surface soil layer. No clear pattern, however, emerged linking these shifts to specific changes in community composition, highlighting the complex interactions between microbial communities, environmental variables, and anthropogenic disturbance. Although several yeast species isolated here have been previously recognized for their biotechnological potential, their diverse and abundant extracellular enzyme profiles were characterized, further highlighting their crucial role in organic matter decomposition and nutrient cycling within the tidal ecosystem, as well as their potential applicability in the food, fine chemical, textile, and pharmaceutical industries.IMPORTANCEThis study presents groundbreaking insights into the yeast diversity of Aoshan Bay, offering invaluable information on their spatial and temporal distribution patterns, as well as their biotechnological potential in the tidal environment. The findings reveal that the eutrophic intertidal flat is a rich repository of yeasts with abundant extracellular enzymatic activity and an important role in nutrient cycling and decomposition processes. Also, these yeasts serve as crucial indicators of ecosystem health and function and are excellent candidates for biotechnological and industrial applications. Collectively, this study not only expands our knowledge of the diversity and distribution of intertidal yeasts but also highlights their promising potential for biotechnological applications.

Plant community composition and traits modulate the impacts of drought intensity on soil microbial community composition and function

Terrestrial ecosystems are increasingly threatened by extreme drought events. Soil microbial communities are central to terrestrial ecosystem function via their role in regulating biogeochemical cycling. Consequently, the impact of increasingly intense drought events on soil microbial communities will have knock-on effects for how ecosystems cope with climate change. In an outdoor grassland mesocosm experiment, we determined how increasing drought intensity affects bacterial and fungal community composition, and functioning, during and after drought. We also tested whether plant community resource acquisition strategy (fast- versus slow-strategy plant communities), plant community composition, and plant functional traits mediate soil microbial responses to increasing drought intensity. We found that increasing drought intensity markedly shifted bacterial and fungal community composition, and these effects persisted until the end of the experiment (two months after re-wetting). Bacterial and fungal communities that experienced severe droughts did not return to baseline composition, while those that experienced a mild drought did. Microbial community functioning (potential extracellular enzyme activity) was reduced at peak drought and shortly after re-wetting. While drought intensity effects on bacterial or fungal communities were insensitive to plant community resource acquisition strategy, functional group abundance (aboveground biomass of grass or forb plant species) composition (grass:forb ratio) and leaf traits (leaf dry matter content and leaf nitrogen concentration) explained significant variation in bacterial and fungal community composition during and after drought. Notably, plant community leaf dry matter content and soil nitrogen were the key factors mediating the effect of increasing drought intensity on microbial indicator taxa (ASVs). We conclude that increasing drought intensity affects grassland soil microbial communities during and after drought, and this impact is influenced by plant community composition and functional traits.

Effects of inorganic nitrogen enrichment on soil CH4 and CO2 production in freshwater and mesohaline marshes across six estuaries in China

Eutrophication is an important environmental stressor in coastal wetlands that alters ecosystem processes including primary production and the carbon cycle. However, how different nitrogen eutrophication scenarios may affect CH4 and CO2 production along a salinity gradient in coastal wetlands is poorly known. We collected the surface soil samples from both tidal freshwater and mesohaline Phragmites australis marshes in six main estuaries in China and conducted inorganic nitrogen enrichment anaerobic incubation experiments. Background soil CH4 and CO2 production rates were strongly correlated with total nitrogen but not salinity. On average, nitrate enrichment decreased soil CH4 and CO2 production rates by ca. 20 % in freshwater marsh and 16–43 % in mesohaline marshes, whereas ammonium enrichment decreased CO2 production by ca. 26–30 % but had no effect on CH4 production. Salinity was not an important mitigating factor in either eutrophication scenario. In most cases, nitrogen addition decreased the extracellular enzyme activities of β-1,4-glucosidase and cellobiohydrolase, but increased the activity of β-N-acetyl glucosaminidase, phenol oxidase and peroxidase. Nitrate addition decreased the mcrA gene abundance on average by 34–35 % but ammonium addition had insignificant effect. Total nitrogen availability was an important driver of CH4 and CO2 production rates in these tidally influenced coastal wetlands. Our results showed that tidal marshes with salinity < 15 ppt had similar soil CH4 production rates, and increasing inorganic nitrogen eutrophication might lower soil microbial carbon gas production in both tidal freshwater and mesohaline marshes.

Fungal and enzymatic pretreatments of bamboo lignocellulose towards high-strength biocomposites: Biological macromolecule/enzyme interaction and bonding enhancement

An energy-intensive and chemical-consuming pretreatment of bamboo is often required to develop its high-performance composites. This study is to evaluate a fungal and enzymatic pretreatment as a sustainable surface modification approach towards high-strength bamboo biocomposites based on D. sinicus. Extracellular enzyme activity analysis suggested that the lignin oxidative enzyme and pectinase secreted by the white-rot fungus T. versicolor achieved higher reaction than the hydrolytic enzymes throughout the pretreatment. Chemical structure analysis revealed that lignin and hemicellulose were partially depolymerised and/or removed during the pretreatment, leading to a relatively higher cellulose content in D. sinicus. The analysis of physical property and microstructure suggested that the surface wettability and porosity of pretreated D. sinicus were significantly improved, facilitating the spreading and penetration of adhesive resins, which subsequently contributed to the superior interfacial bonding of the biocomposites. The tensile strength of the biocomposites increased from 195.8 MPa to 245.8 MPa and the bonding strength increased from 4.9 MPa to 6.3 MPa after the pretreatment, which were 1.26 times and 1.28 times those of the untreated biocomposite respectively. The results proved that the fungal and enzymatic pretreatment is a promising, eco-friendly and efficient surface modification approach for the preparation of high-strength lignocellulosic biocomposites.

Isolation of Heavy Metal-Tolerant and Anti-Phytopathogenic Plant Growth-Promoting Bacteria from Soils.

In this study, by isolating multifunctional soil bacteria that can promote plant development, resist heavy metals, exhibit anti-phytopathogenic action against plant diseases, and produce extracellular enzymes, we hope to improve the effectiveness of phytoremediation techniques. To isolate multifunctional soil bacteria, we used soils with diverse characteristics as isolation sources. To look into the diversity and structural traits of the bacterial communities, We conducted amplicon sequencing of the 16S rRNA gene on five types of soils and predicted functional genes using Tax4Fun2. The isolated bacteria were evaluated for their multifunctional capabilities, including heavy metal tolerance, plant growth promotion, anti-phytopathogenic activity, and extracellular enzyme activity. The genes related to plant growth promotion and anti-phytopathogenic activity were most abundant in forest and paddy soils. Burkholderia sp. FZ3 and FZ5 demonstrated excellent heavy metal resistance (≤ 1 mM Cd and ≤ 10 mM Zn), Pantoea sp. FC24 exhibited the highest protease activity (24.90 μmol tyrosine·g-DCW-1·h-1), and Enterobacter sp. PC20 showed superior plant growth promotion, especially in siderophore production. The multifunctional bacteria isolated using traditional methods included three strains (FC24, FZ3, and FZ5) from the forest and one strain (PC20) from paddy field soil. These results indicate that, for the isolation of beneficial soil microorganisms, utilizing target gene information obtained from isolation sources and subsequently exploring target microorganisms is a valuable strategy.

Arbuscular mycorrhizal symbiosis enhances the accumulation of plant-derived carbon in soil organic carbon by regulating the biosynthesis of plant biopolymers and soil metabolism

Plant-derived carbon (C) is a critical constituent of particulate organic carbon (POC) and plays an essential role in soil organic carbon (SOC) sequestration. Yet, how arbuscular mycorrhizal fungi (AMF) control the contribution of plant-derived C to SOC storage through two processes (biosynthesis of plant biopolymers and soil metabolism) remains poorly understood. Here, we utilized transcriptome analysis to examine the effects of AMF on P. communis roots. Under the AM symbiosis, root morphological growth and tolerance to stress were strengthened, and the biosynthetic pathways of key plant biopolymers (long-chain fatty acids, cutin, suberin, and lignin) contributing to the plant-derived C were enhanced. In the subsequent metabolic processes, AMF increased soil metabolites contributing to plant-derived C (such as syringic acid) and altered soil metabolic pathways, including carbohydrate metabolism. Additionally, C-acquiring soil extracellular enzyme activities were enhanced by AMF, which could affect the stabilization of plant-derived C in soil. The contents of POC (21.71 g kg−1 soil), MAOC (10.75 g kg−1 soil), and TOC (32.47 g kg−1 soil) in soil were significantly increased by AMF. The concentrations of plant-derived C and microbial-derived C were quantified based on biomarker analysis. AMF enhanced the content of plant-derived C in both POC and MAOC fractions. What's more, plant-derived C presented the highest level in the POC fraction under the AMF treatment. This research broadens our understanding of the mechanism through which plant-derived C contributes to the accumulation of POC and SOC induced by AM symbiosis, and evidences the benefits of AMF application in SOC sequestration.

When are you measuring soil β‐glucosidase activities in cropping systems?

AbstractIn situ soil respiration is driven by annual patterns of temperature and soil moisture, but what about extracellular enzyme activities responsible for depolymerizing soil organic matter? We conducted biweekly measurements of potential soil β‐glucosidase activities during a 4‐month period from March soil thawing through July in annually cropped field plots in eastern South Dakota. Our objective was to determine the best sampling time to resolve the effects of crop rotational diversity on soil microbial activities. Potential β‐glucosidase activities were elevated immediately following soil thaw, peaked in May, and declined to their lowest value in mid‐summer. Temperature and precipitation had no value in predicting enzyme activities; however, enzyme activities were affected by crop rotational diversity and responded to current crop and previous crop. These findings are pertinent to the use of soil extracellular enzymes in soil health assessments and as indicators of microbial substrate preference with implications for soil carbon stabilization.

Microplastics impair extracellular enzymatic activities and organic matter cycling in oligotrophic sandy marine sediments

Microplastics (MPs) are ubiquitous and constantly accumulating in the marine environment, especially sediments. Yet, it is not well clarified if and how their carbon backbone could interact with surrounding sediments, eventually impairing key benthic processes. We assessed the effects of a ‘pulse’ contamination event of MPs on sedimentary organic matter (OM) quantity, quality and extracellular enzymatic activities (EEAs), which are well established descriptors of benthic ecosystem functioning. Marine sediments were exposed for 30 days to environmentally relevant concentrations (∼0.2 % in weight) of naturally weathered particles (size range 70–210 μm) of polyurethane, polyethylene, and a mixture of the most common polymers that are documented to accumulate in marine sediments. Despite the low concentration, contaminated sediments showed significantly different composition of OM, showing a decrease in lipid content and increase in protein. Moreover, we document a significant decrease (over 25 %) in quantity of biopolymeric C already after 15 days of exposure, compared to controls. Contaminated sediments showed lower C degradation rates (up to −40 %) and altered EEAs, with alkaline phosphatase being ∼50 % enhanced and aminopeptidase being reduced over 35 % compared to control treatments. Overall, the effects generated by the mixture of polymers were smaller than those exerted by the same amount of a single polymer. Our results provide insights on how that MPs can significantly alter marine sedimentary biogeochemistry through altered benthic processes, that could cumulatively impair whole benthic trophic webs by enhancing the accumulation and possible longer-term storage of recalcitrant organic C in the seabed.

The Responses of Soil Extracellular Enzyme Activities and Microbial Nutrients to the Interaction between Nitrogen and Phosphorus Additions and Apoplastic Litter in Broad-Leaved Korean Pine Forests in Northeast China

The impact of nitrogen and phosphorus deposition alternations, as well as apoplastic litter quality and quantity, on soil nutrient cycling and soil carbon pool processes in forest ecosystems is of considerable importance. Soil ecological enzyme chemistry is a powerful tool for elucidating the nutrient limitations of microbial growth and metabolic processes. In order to explore the responding mechanisms of soil ecological enzyme chemistry to the simultaneous changes in apoplast input and nitrogen and phosphorus deposition in temperate coniferous and broad-leaved mixed forests, an outdoor simulating experiment was conducted. The results demonstrate that the treatments involving apoplastic material and nitrogen and phosphorus additions had significantly impacted soil nutrient levels across different forest types. Apoplastic treatments and N-P additions had a significant effect on the soil total organic carbon (TOC), dissolved organic carbon (DOC), soil total soluble nitrogen (TSN), soil available phosphorus (SAP), soil total nitrogen (TN), soil total phosphorus (TP), and microbial biomass carbon (MBC). However, the effects on soil microbial biomass (MBN) and microbial biomass phosphorus (MBP) were insignificant. The apomictic treatments with N and P addition did not result in a statistically significant change in soil C-hydrolase activities (β-1,4-glucosidase BG, β-1,4-xylosidase BX, cellobiohydrolase CBH, phenol oxidase POX, and peroxidase PER), N-hydrolase activities (β-1,4-N-acetylglucosaminidase NAG and L-leucine aminopeptidase LAP), or P-hydrolase activities (Acid phosphatase AP). Although the apomictic treatments did not yield a significant overall impact on carbon hydrolase activity, they influenced the activity of specific enzymes, such as CBH, LAP, and PER, to varying degrees. The effects on BG, BX, CBH, AP, and C-hydrolase activities were significant for different stand types. The impact of apomictic treatments and N-P additions on soil nitrogen hydrolase activities was inconsequential with a minimal interactive effect. The highest correlation between PER, LAP, and N-hydrolase activities was observed in conjunction with elevated levels of nitrogen and phosphorus addition (N3L0, original litter treatment, and high amounts of N and P addition). These findings may provide a theoretical foundation for the management of ecosystem function in broad-leaved Korean pine forests.

“Efficient novel fungal-enriched biochar formulation for hexavalent chromium bioremediation”

Chromium (Cr), a key element in industrial processes such as leather tanning, poses severe environmental hazards, particularly its hexavalent form, Cr(VI), which is highly toxic and prevalent in tannery effluents/sludge. The persistence and toxicity of Cr(VI) necessitate the development of effective remediation strategies to mitigate its environmental impact. This study investigated the potential of Trichoderma yunnanense (NBRICRF_97) and its combination with 0.5% sugarcane bagasse biochar (SBC) for the reduction of Cr(VI). The results demonstrated that T. yunnanense alone achieved a 91.04% reduction of 50 mg L−1 Cr(VI) within 72 h. Combined with 0.5% SBC, the reduction efficiency increased to 99.65% within 48 h. However, the efficiency decreased at higher concentrations (200 mg L−1). The combination also improved fungal growth and increased extracellular ChrR enzyme activity (13.07 U mg−1 protein compared to the control). Total glutathione activity was boosted by 161.07% at 100 mg L−1 Cr(VI). Antioxidant enzymes (SOD, POD, CAT) and proline mitigated oxidative stress and FTIR analysis revealed changes in fungal cell wall functional groups (-OH and -NH) upon Cr(VI) exposure. SEM-EDX confirmed chromium deposition on fungal surfaces. These results underscore the Cr(VI) detoxification capabilities of T. yunnanense and the synergistic benefits of SBC, suggesting a promising bioremediation strategy for Cr(VI)-contaminated environments. The integration of T. yunnanense with SBC offers a sustainable and cost-effective approach for the bioremediation of Cr(VI)-contaminated sites, with potential for implementation in large-scale environmental cleanup efforts.

Dominant Tree Species and Litter Quality Govern Fungal Community Dynamics during Litter Decomposition.

Litter decomposition is a crucial biochemical process regulated by microbial activities in the forest ecosystem. However, the dynamic response of the fungal community during litter decomposition to vegetation changes is not well understood. Here, we investigated the litter decomposition rate, extracellular enzyme activities, fungal community, and nutrient cycling-related genes in leaf and twig litters over a three-year decomposition period in a pure Liquidamabar formosana forest and a mixed L. formosana/Pinus thunbergii forest. The result showed that during the three-year decomposition, twig litter in the mixed forest decomposed faster than that in the pure forest. In both leaf litter and twig litter, β-cellobiosidase and N-acetyl-glucosamidase exhibited higher activities in the mixed forest, whereas phosphatase, β-glucosidase, and β-xylosidase were higher in the pure forest. The fungal α-diversity were higher in both litters in the pure forest compared to the mixed forest, with leaf litter showing higher α-diversity than twig litter. Fungal species richness and α-diversity within leaf litter increased as decomposition progressed. Within leaf litter, Basidiomycota dominated in the mixed forest, while Ascomycota dominated in the pure forest. Funguild analysis revealed that Symbiotroph and ectomycorrhizal fungi were more abundant in the mixed forest compared to the pure forest. In the third-year decomposition, genes related to phosphorus cycling were most abundant in both forests, with the pure forest having a higher abundance of cex and gcd genes. Fungal community structure, predicted functional structure, and gene composition differed between the two forest types and between the two litter types. Notably, the fungal functional community structure during the first-year decomposition was distinct from that in the subsequent two years. These findings suggest that dominant tree species, litter quality, and decomposition time all significantly influence litter decomposition by attracting different fungal communities, thereby affecting the entire decomposition process.

Strong large‐scale structure–function coupling in benthic bacteria is mediated by algae in a geodiverse river network

AbstractBenthic bacteria in stream ecosystems drive organic matter mineralization. However, knowledge on how this ecosystem function is driven by bacterial community composition in interaction with environmental conditions and organic matter resources is poor. This is especially true when considering the regional scale of river networks, at which environmental conditions vary in a scale‐dependent manner and are spatially structured due to asymmetrical water flow. Similarly, organic matter resources may have a terrestrial origin in remote headwaters or be sourced locally from algae living in close proximity to bacteria in benthic biofilms. We investigated benthic biofilm meta‐community structure and function across the &gt; 6700 km2 river network of the near‐natural Vjosa in Albania and Greece and found a strong control of the benthic algal community on bacterial community composition (13.4% of variability explained). In addition, bacterial community composition has linkages to water chemistry, which itself is strongly shaped by the diverse geology in the catchment, and to dispersal, shaping metacommunity structure as a neutral process. Notably, bacterial community composition explained the largest single fraction of variability (31.5%) in extracellular enzymatic activities, while there was no dependency of enzyme ratios on organic matter nor environmental conditions. Synergistic effects between bacteria and algae accounted for additional 47.3% of variability in heterotrophic functioning, emphasizing the importance of algal–bacterial interactions in benthic biofilms. Our findings shed new light on bacterial structure–function coupling highlighting the importance of algal–bacterial interactions at the river network scale.

Deep return of straw improves topsoil health and maize productivity more efficiently than shallow return

AbstractStraw return is an important nature‐based solution to identify optimal management practices to maintain cropland soil health and food security. However, there is still limited information available regarding the impacts of different straw return strategies on soil health, especially when considering a comprehensive assessment of soil physical, chemical and biological characteristics. Soil microorganisms are the core and key to maintaining soil health. The present research aims to determine the impact of straw return to different depths on soil health index (SHI) and crop productivity across different soil layers. We conducted a 3–year field experiment with four treatments: a blank control with no straw return (NR), straw return treatments with shallow return (SR), mulch return (MR) and deep return (DR), and the straw application rate was 10,500 kg/ha. The results showed that it was the microbial indicators that dominated the effects on soil health, rather than soil organic carbon (SOC). Specifically, we found that the SR treatment exhibited the highest extracellular enzyme activities of both hydrolases and oxidases in the subsoil, and the peroxidase (PEO) activity was 758.99 nmol/g/h. In addition, compared with MR and DR treatments, the abundance of symbiotroph fungi in SR treatment increased by 78.41% and 54.38%, releasing more nutrients to the soil. Hence, SR treatment increased SHI mainly by increasing the PEO activity and NO3−–N content, and SR treatment significantly increased SHI by 25.59% and 22.16% compared with MR and DR treatments. By contrast, the DR treatment greatly increased the topsoil SHI by 43.33% and 58.98% compared with SR and SM treatments, mainly due to the enhanced microbial biomass N (MBN) and dissolved organic nitrogen (DON). The DR treatment in topsoil had the lowest soil bulk density (BD) which enhanced Bacteroidota but decreased Methylomirabilota abundance, thus resulting in the accumulation of MBN. Moreover, DR treatment led the highest maize yield which increased with an increasing SHI. To conclude, DR treatment is an effective method to improve SHI and crop productivity in nutrient‐rich topsoil, while SR treatment has a much better effect in the subsoil. Our results will provide a theoretical paradigm for selecting straw return strategies, which is crucial for promoting soil health and food productivity, achieving high crop yields, and supporting sustainable agricultural development.

The legacy effects of mycorrhizal fungal inoculation alleviate soil nitrogen limitations by modulating the homeostasis of extracellular enzymes and fungal function in arid mining regions

AbstractRe‐vegetation types and the application of arbuscular mycorrhizal fungi (AMF) play a crucial role in regulating soil conditions and enhancing vegetation cover during ecological restoration in the northern arid mining area. However, the impact of AMF on the balance of soil–plant–microbial ecosystems through its influence on microbial activities remains unclear. Therefore, we studied the influence of AMF on soil nutrient limitation and the soil fungal community in five different vegetation types in the Daliuta reclamation area. Through the measurement of soil extracellular enzyme activity and the analysis of soil fungal communities by high‐throughput sequencing, we found that the ratio of lnBG:ln(NAG + LAP):lnAP in the AMF sample was closer to 1:1:1, and the soil ecological enzyme stoichiometry was more balanced. The results of enzyme vector showed that AMF inoculation could alleviate the restriction of soil nitrogen nutrients. The results of high‐throughput sequencing showed that AMF improved the diversity of soil fungal community by regulating the composition of soil fungal community, and finally alleviated the nitrogen restriction during decomposition. Functional prediction of soil fungi using FUNGuild indicated that the soil fungi present were mainly saprophytic, saprophytic–symbiotic, and pathotroph–saprophytic–symbiotic trophic types. AMF reduced the percentage of pathotroph fungi and also tended to transform saprophytic into saprophytic–symbiotic type. The findings suggest that mycorrhizal reclamation can enhance the homeostasis of extracellular enzyme dynamics by optimizing fungal community structure and modulating fungal functionality, thereby alleviating microbial nutrient constraints and improving plant adaptability to arid environments. This article provides a theoretical underpinning for eco‐friendly mining construction and sustainable ecological development in arid mining areas.

Ectomycorrhizal fungi and the nitrogen economy of Nothofagus in southern Patagonia.

Subantarctic Nothofagus forests are the southernmost forests in the world, with negligible atmospheric nitrogen (N) deposition. Most paradigms about the role of ectomycorrhizal (ECM) fungi in N cycling and plant N uptake at high latitudes have been tested in boreal coniferous forests, while in the southern hemisphere, ECM hosts are primarily angiosperms. Using ITS1 meta-barcoding, we characterized ECM and saprotrophic fungal communities in evergreen and deciduous Nothofagus forests forming monodominant and mixed stands in the archipelago of Tierra del Fuego (Chile and Argentina). We assessed the N economy of Nothofagus by correlating host species with fungal relative abundances, edaphic variables, net N mineralization, microbial biomass N and the activity of eight extracellular soil enzymes activities. The N economy of deciduous N. pumilio forests was strikingly similar to boreal coniferous forests, with the lowest inorganic N availability and net N mineralization, in correlation to higher relative abundances of ECM fungi with enzymatic capacity for organic N mobilization (genus Cortinarius). In contrast, the N economy of evergreen N. betuloides forests was predominantly inorganic and correlated with ECM lineages from the family Clavulinaceae, in acidic soils with poor drainage. Grassy understory vegetation in deciduous N. antarctica forests likely promoted saprotrophic fungi (i.e., genus Mortierella) in correlation with higher activities of carbon-degrading enzymes. Differences between Nothofagus hosts did not persist in mixed forests, illustrating the range of soil fertility of these ECM angiosperms and the underlying effects of soil and climate on Nothofagus distribution and N cycling in southern Patagonia.

Peptide-Driven Assembly of Magnetic Beads for Potentiometric Sensing of Bacterial Enzyme at a Subcellular Level.

Bacterial enzymes with different subcellular localizations play a critical ecological role in biogeochemical processing. However, precisely quantifying enzymes localized at certain subcellular levels, such as extracellular enzymes, has not yet been fully realized due to the complexity and dynamism of the bacterial outer membrane. Here we present a magneto-controlled potentiometric sensing platform for the specific detection of extracellular enzymatic activity. Alkaline phosphatase (ALP), which is one of the crucial hydrolytic enzymes in the ocean, was selected as the target enzyme. Magnetic beads functionalized with an ALP-responsive self-assembled peptide (GGGGGFFFpYpYEEE, MBs-peptides) prevent negatively charged peptides from entering the bacterial outer membrane, thereby enabling direct potentiometric sensing of extracellular ALP both attached to the bacterial cell surface and released into the surrounding environment. The dephosphorylation-triggered assembly of peptide-coupled magnetic beads can be directly and sensitively measured by using a magneto-controlled sensor. In this study, extracellular ALP activity of Pseudomonas aeruginosa at concentrations ranging from 10 to 1.0 × 105 CFU mL-1 was specifically and sensitively monitored. Moreover, this magneto-controlled potentiometric method enabled a simple and accurate assay of ALP activity across different subcellular localizations.

Release of intracellular enzymes increases the total extracellular activities of carbohydrate processing enzymes in marine environment.

Microbial extracellular enzymatic activities (EEAs) produced by microbes to degrade biopolymers are the 'gatekeeper' of carbon cycle in the marine ecosystem. It is usually assumed that these extracellular enzymes are actively secreted by microbes. But biopolymers degrading enzymes also exist in the intracellular space. Cell lysis will passively release these enzymes into the environments and contribute to the total EEAs. However, to what extent the cell lysis can contribute to the total EEAs are still unclear. Here, using extreme cell lysis method, we evaluated the maximum contribution of cell lysis to total EEAs in culturable marine bacteria and coastal seawater. For carbohydrate processing enzymes (β-glucosidase, alginate lyase and chitinase), the release of intracellular enzymes could contribute positively (up to 56.1% increase for β-glucosidase in seawater) to the total EEAs. For protease and leucine aminopeptidase, the cell lysis did not increase and even decreased the total EEAs. For alkaline phosphatase, the intracellular enzymes generally had no contribution to the total EEAs. These results showed that passively released intracellular enzymes could substantially increase the total extracellular activities of carbohydrate processing enzymes, which should be considered in building the link between the EEAs and organic carbon cycle in the ocean.

Nutrient regeneration patterns for initiating and maintaining algae blooms-a case study of in Lake Taihu

In order to clarify the nitrogen (N) and phosphorus (P) regeneration patterns and internal mechanism for initiating and maintaining algal blooms in Lake Taihu, samples (including surface water and sediment) from 8 sites in Lake Taihu were collected for nine times from May 2010 to April 2011, and analyzed for total and labile organic matter, P fractionation and sorption behaviors, extracellular enzymatic activities (EEA), dehydrogenase activity, the respiratory electron transport system activity, and iron in sediment, EEA, N and P species and chlorophyll a (Chl. a) in surface water, as well as N and P species in interstitial water. In Lake Taihu, although severe blooms occurred in both Meiliang Bay and Zhushan Bay, the nutrient regeneration patterns stimulating the initiation and maintenance of algae blooms in these two bays were different. In Zhushan Bay with low EEA in surface water, abundant N and P flux from sediments, due to the degradation of organic matter and enzymatic hydrolysis in sediment, further stimulated the initiation and maintenance of algae blooms. In Meiliang Bay, in spite of lower nutrient supply from sediment, high EEA in surface water occurred later than the serious blooms, showing that the nutrient regeneration from sediment, not water body, was still the trigger for the start of the bloom, and sediment nutrient release and predominant surface water nutrient regeneration by abundant exoenzymes sustained the algal blooms. In the Western region, algal bloom started in the northern area and further spread in the remaining part of the lake; nutrient regeneration in the surface water sustained the slight bloom. In the East Bays, the decay and decomposition of macrophytes led to anaerobic conditions in sediments and high ammonia in interstitial water, but low iron bound phosphorus resulted in anaerobic release of very few P, thus showed extremely low Chl. a concentration.