Extracellular Enzymes Research Articles

Streptomyces as a Novel Biotool for Azo Pigments Remediation in Contaminated Scenarios.

Azo pigments are widely used in the textile and leather industry, and they generate diverse contaminants (mainly in wastewater effluents) that affect biological systems, the rhizosphere community, and the natural activities of certain species. This review was performed according to the Systematic Reviews and Meta Analyses (PRISMA) methodology. In the last decade, the use of Streptomyces species as biological azo-degraders has increased, and these bacteria are mainly isolated from mangroves, dye-contaminated soil, and marine sediments. Azo pigments such as acid orange, indigo carmine, Congo red, and Evans blue are the most studied compounds for degradation, and Streptomyces produces extracellular enzymes such as peroxidase, laccase, and azo reductase. These enzymes cleave the molecule through asymmetric cleavage, followed by oxidative cleavage, desulfonation, deamination, and demethylation. Typically, some lignin-derived and phenolic compounds are used as mediators to improve enzyme activity. The degradation process generates diverse compounds, the majority of which are toxic to human cells and, in some cases, can improve the germination process in some horticulture plants. Future research should include analytical methods to detect all of the molecules that are generated in degradation processes to determine the involved reactions. Moreover, future studies should delve into consortium studies to improve degradation efficiency and observe the relationship between microorganisms to generate scale-up biotechnological applications in the wastewater treatment industry.

Process and mechanism of termite impact on soil and plant.

Termites, as a kind of nesting social insects, are often confused as worldwide "pests" because some of their groups have great destructive effects. The vast majority of termites can regulate ecosystem functions and ser-vices by participating in biogeochemical cycles, known as "ecosystem engineers". We reviewed studies on the effects of termites on the physical, chemical and biological characteristics of mound soil ecosystems and the composition and diversity of plant communities. Termites could form unique soil "biogenic aggregates" and "resource heterogeneity patches", which affect microbial community structure, extracellular enzyme activity, physicochemical property and greenhouse gas emission, thereby affecting plant growth, community composition and structure, and vegetation productivity. However, this effect significantly differed among termite groups and functional groups, and was dependent on regional soil environment and microclimate conditions. Meanwhile, termite-mound could effectively improve ecosystem adaptation or resistance to environmental stress through the above process. Future research should focus on the following directions: 1) studying the trophic cascading effect of termite-centered soil multilevel biological network and the potential effect on biogeochemical cycle from microscale (aggregate level) to macroscale (landscape level); 2) exploring the potential of termite mound soil as a fertility amendment in tropical regions, and mining beneficial microbial functional genes to develop related products for termite control.

Discovering the role of fairy ring fungi in accelerating nitrogen cycling to promote plant productivity in grasslands

Soil microorganisms play a key role in the provision of plant-bioavailable nutrients, which is crucial for ecosystem functioning and plant productivity. Fairy rings are widespread features in grasslands accompanied by lush dark-green vegetation bands. This enigmatic feature is caused by increased soil bioavailable nitrogen (N) due to the expansion of fairy ring fungi (FRF). However, little is known about how FRF enhance soil bioavailable N concentrations. Here, we conducted a survey of 35 fairy rings in temperate grasslands to reveal the role of FRF in regulating soil microorganisms and N cycling using amplicon and metagenomic sequencing. The presence of FRF accelerated organic N mineralization via promoting extracellular enzyme (β-1,4-N-acetylglucosaminidase) activity, leading to a 455% increase in ammonium-N. This further stimulated nitrification to enhance nitrate-N concentration by favoring ammonia-oxidizing archaea. Concomitantly, the increased nitrate-N did not promote denitrification or affect the potential risk of N loss. Furthermore, the relative abundance of other saprotrophic and symbiotrophic fungi was significantly reduced by FRF but the changes in these fungi did not affect the activity of extracellular enzymes involved in N mineralization. Our results suggest that FRF can act as ecosystem engineer species shaping fairy rings by driving soil N cycling without the involvement of other microbial functional groups of saprotroph and symbiotroph to boost plant productivity. Thus, due to the stronger N mobilizing ability, FRF show great potential to be exploited as beneficial microorganisms in plant production and sustainable agricultural development.

Isolation and characterization of a novel bacterium that promotes the degradation of poly(glycolic acid) by its extracellular esterase under thermophilic conditions

Poly(glycolic acid) (PGA) is widely utilized in the shale oil and gas industry owing to its biodegradable nature, as well as superior mechanical and barrier properties. However, PGA degradability is limited by environmental conditions such as temperature and pH, and using acids to optimize the degradation can have adverse environmental impact. Thus, it is essential to identify methods that can effectively promote the degradation of PGA under mild conditions, such as microbial degradation. In the present study, strain DB14, a bacterium that promotes PGA degradation, was isolated from drain water of a steam pipeline. Sequence analysis of the 16S rRNA gene of the bacterium revealed that it is mostly closely related to Geobacillus icigianus; however, it also differed from Geobacillus icigianus in several physiological properties. To investigate PGA degradation by strain DB14, a PGA film and disc were incubated with the strain. The residual weight of the PGA film (thickness: 170 μm) significantly reduced after incubation, whereas the decrease in the thickness of the PGA disc (thickness: 3 mm) was relatively small. The penetration of water, the bacterium, and the extracellular enzymes into the interior from the reaction-erosion front of the PGA disc may be inhibited by the high barrier performance of PGA. Strain DB14 was also found to change the pH of the surrounding environment to approximately 8–9. To investigate the effect of pH on PGA degradability, degradation tests with crude extracellular enzymes derived from strain DB14 were conducted in various buffers. The results showed that the degradation activity was highest at pH 8, which implied that DB14 efficiently maximized the hydrolytic capacity of its enzyme for degrading PGA. Thus, this study provides a basis for developing environmentally friendly technologies that can promote the degradation of PGA molding articles, especially those used in wellbores for oil and gas recovery.

Inhibition of diastatic yeasts by Saccharomyces killer toxins to prevent hyperattenuation during brewing.

Secondary fermentation in beer can result in undesirable consequences, such as off-flavors, increased alcohol content, hyperattenuation, gushing, and the spontaneous explosion of packaging. Strains of Saccharomyces cerevisiae var. diastaticus are a major contributor to such spoilage due to their production of extracellular glucoamylase enzyme encoded by the STA1 gene. Saccharomyces yeasts can naturally produce antifungal proteins named "killer" toxins that inhibit the growth of competing yeasts. Challenging diastatic yeasts with killer toxins revealed that 91% of strains are susceptible to the K1 killer toxin produced by S. cerevisiae. Screening of 192 killer yeasts identified novel K2 toxins that could inhibit all K1-resistant diastatic yeasts. Variant K2 killer toxins were more potent than the K1 and K2 toxins, inhibiting 95% of diastatic yeast strains tested. Brewing trials demonstrated that adding killer yeast during a simulated diastatic contamination event could prevent hyperattenuation. Currently, most craft breweries can only safeguard against diastatic yeast contamination by good hygiene and monitoring for the presence of diastatic yeasts. The detection of diastatic yeasts will often lead to the destruction of contaminated products and the aggressive decontamination of brewing facilities. Using killer yeasts in brewing offers an approach to safeguard against product loss and potentially remediate contaminated beer.IMPORTANCEThe rise of craft brewing means that more domestic beer in the marketplace is being produced in facilities lacking the means for pasteurization, which increases the risk of microbial spoilage. The most damaging spoilage yeasts are "diastatic" strains of Saccharomyces cerevisiae that cause increased fermentation (hyperattenuation), resulting in unpalatable flavors such as phenolic off-flavor, as well as over-carbonation that can cause exploding packaging. In the absence of a pasteurizer, there are no methods available that would avert the loss of beer due to contamination by diastatic yeasts. This manuscript has found that diastatic yeasts are sensitive to antifungal proteins named "killer toxins" produced by Saccharomyces yeasts, and in industrial-scale fermentation trials, killer yeasts can remediate diastatic yeast contamination. Using killer toxins to prevent diastatic contamination is a unique and innovative approach that could prevent lost revenue to yeast spoilage and save many breweries the time and cost of purchasing and installing a pasteurizer.

Effect of Solid-State Fermentation of Hericium erinaceus on the Structure and Physicochemical Properties of Soluble Dietary Fiber from Corn Husk.

Corn husk, a by-product of corn starch production and processing, contains high-quality dietary fiber (DF). Our study compares and analyzes the impact of Hericium erinaceus solid-state fermentation (SSF) on the structure and physicochemical characteristics of soluble dietary fiber (SDF) of corn husks. The study also investigates the kinetics of SSF of H. erinaceus in this process. The scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT-IR) results revealed significant structural changes in corn husk SDF before and after fermentation, with a significant elevation in the functional group numbers. The data indicate that the fermented corn husk SDF's water-holding, swelling, and oil-holding capacities increased to 1.57, 1.95, and 1.80 times those of the pre-fermentation SDF, respectively. Additionally, the results suggest that changes in extracellular enzyme activity and nutrient composition during SSF of H. erinaceus are closely associated with the mycelium growth stage, with a mutual promotion or inhibition relationship between the two. Our study offers a foundation for corn husk SDF fermentation and is relevant to the bioconversion of maize processing by-products.

Resin Acid Copper Salt, an Interesting Chemical Pesticide, Controls Rice Bacterial Leaf Blight by Regulating Bacterial Biofilm, Motility, and Extracellular Enzymes.

Bacterial virulence plays an important role in infection. Antibacterial virulence factors are effective for preventing crop bacterial diseases. Resin acid copper salt as an effective inhibitor exhibited excellent anti-Xanthomonas oryzae pv. oryzae (Xoo) activity with an EC50 of 50.0 μg mL-1. Resin acid copper salt (RACS) can reduce extracellular polysaccharides' (EPS's) biosynthesis by down-regulating gumB relative expression. RACS can also effectively inhibit the bio-mass of Xoo biofilm. It can reduce the activity of Xoo extracellular amylase at a concentration of 100 μg mL-1. Meanwhile, the results of virtual computing suggested that RACS is an enzyme inhibitor. RACS displayed good curative activity with a control effect of 38.5%. Furthermore, the result of the phytotoxicity assessment revealed that RACS exhibited slight toxicity compared with the control at a concentration of 200 μg mL-1. The curative effect was increased to 45.0% using an additional antimicrobial agent like orange peel essential oil. RACS markedly inhibited bacterial pathogenicity at a concentration of 100 μg mL-1 in vivo.

Effects of farmyard manure and chemical fertilizer application rates on soil biology, cotton and fiber yield

Organic and inorganic fertilizers have significant effect on plant physiology, yield per unit area, available plant nutrient contents and extracellular enzyme activities of soils. This study was carried out in field conditions in arid and semi-arid regions between 2020-2021 years, May 01. The effects of farmyard manure (FM) (20, 40, 60 Mg ha-1) and chemical (CF) (350 kg urea ha-1, 100, 200, 300 kg DAP ha-1) fertilizers applied at different rates on plant nutrient (N, P, K, Ca, Mg, Fe, Cu, Zn and Mn) contents, SPAD value and NDVI of cotton plants, seed cotton yield and soil enzymes (urease, catalase, dehydrogenase, alkaline phosphatase) were investigated. The results showed that FM applications significantly (p<0.01) increased the plant macro and micronutrients compared to CF applications, except for N (200 + 150 kg urea ha-1), Zn and Cu (300 kg DAP + 200 + 150 kg urea ha-1) in the 2021 cotton growing season. Mineralization of FM is slow under natural conditions; therefore, the use of FM alone is not sufficient to meet the nutrient needs of high yielding varieties. Urease and dehydrogenase activities increased significantly in FM treated soils compared to CF, while no significant (p<0.01) increase was recorded in alkaline phosphatase and catalase activities. Farmyard manure is a useful management practice for increasing soil biological activity. Physiological parameters of NDVI, SPAD and seed cotton yield significantly increased in FM treated soils compared to CF applications. The increase in cotton yield was 29.15%, in NDVI value was 22.38% and in SPAD value was 121.7%. The main issues with cotton in the area are the low organic carbon content of the soils, high clay content, arid and semi-arid soils, and their detrimental impact on the uptake of particular nutrients (N, P and B).

Silkworm Gut Microflora as a Source: Biotechnological Applications

The term "microflora" has gained popularity as researchers explore the symbiotic relationships between microorganisms and their hosts, from our skin to our gut. Insects, including the silkworm, have mutualistic ties with their gut microflora, making them a valuable resource for studying essential molecules. The silkworm, with its rapid generation time, abundant genetic resources, well-defined genetic background, and numerous homologous genes, is an ideal model organism for various life science studies. Silkworm rearing is simple, inexpensive, and does not require ethical clearance, allowing the gut microflora of Bombyx mori to be harnessed for biotechnological applications. Silkworm gut microbes can synthesize and produce extracellular enzymes, such as cellulases, proteases, and lipases, which are crucial for industries ranging from biofuel production to waste management and textile processing. Additionally, these microbes can produce vitamins, such as vitamin B12, and metabolites like short-chain fatty acids (SCFAs), which have potential applications in the food and pharmaceutical industries. Antimicrobial substances produced by the gut microflora, including bacteriocins and lactic acid, are of particular therapeutic importance, offering natural alternatives to traditional antibiotics and contributing to the development of novel antimicrobial therapies. Moreover, the antioxidants generated by these symbionts, such as glutathione and superoxide dismutase (SOD), have significant implications for health supplements and cosmetics, owing to their ability to combat oxidative stress and support immune function. Beyond their metabolic capacities, silkworm symbionts can be used to enhance health, target and manage agricultural pests, and control vectors of human diseases in an environmentally friendly manner. Recent research has also highlighted the potential of mutualistic symbiotic bacteria for plastic disposal, with enzymes like PETases showing promise in degrading synthetic polymers. Despite extensive knowledge on the biology and physiology of the silkworm Bombyx mori, few studies have focused on its gut microflora. Modern genetic techniques, such as the genetic transformation of silkworms with genes of interest, may help overcome these research gaps. Increasing research efforts on silkworm gut microflora will not only reveal new symbiotic relationships but also identify new sources of biotechnologically important molecules and enzymes for therapeutic and industrial applications.

Decolorization of Textile Dyes by Extracellular Enzymes Produced from Trametes sanguinea and Perenniporia taephropora Immobilized on Natural Media

The color of textile wastewater is still a main problem in wastewater treatment by biological processes. The colored effluents from textile factories usually exceed effluent standards. Therefore, various innovations were developed to treat textile wastewater for decolorization in the effluents. This research aims to decolorize textile wastewater by immobilizing white rot fungi degradation. At first, the 11 fungal stains were tested to find the decolorized efficiency then the high decolorized efficiency fungal stains were immobilized on four material media, namely water hyacinth stalks, coconut husk, corn cob, and loofah. After that, the immobilized fungi were cultivated in the culture media at 30, 60, and 120 C/N ratios, respectively. The results showed that Trametes sanguinea and Perenniporia tephropora were two stains with a high decolorized efficiency of 68.8% and 67.5% respectively, and the decolorized efficiency was increased when immobilized on loofahs and fed with 120 C/N ratio medium. In a comparison of two fungal stains, P. tephropora was found more suitable for the decolorization of textile wastewater than T. sanguinea because T. sanguinea could produce red-orange pigments that induced the colored enhancement in wastewater over time. Finally, immobilized P. tephropora was cultivated in a 120 C/N ratio medium within a 10 L continuous stirred tank reactor (8 L working volume) to investigate the decolorized efficiency, enzymatic activity, and repeated batch. It was found that three repeated cycles were carried out by reusing the immobilized P. tephropora and the highest decolorized efficiency was 63.4%. The enzymatic activity of laccase, manganese peroxidase, and lignin peroxidase was 15.5 U/L, 85.9 U/L, and 0 U/L, respectively

Smooth vetch covering alters soil aggregate microbial metabolic limitations in citrus orchards

Green manure covering alters the supply-demand relationship between soil resources and microorganisms by improving soil structure and increasing carbon inputs. However, it remain unclear how soil microorganism respond to the imbalance between resources and demands at the aggregate scale. The present study analyzed the stoichiometric ratios of organic carbon, nitrogen, and phosphorus nutrients, microbial extracellular enzyme activities, and biomass in large macroaggregates (LMA, 2–8 mm), small macroaggregates (SMA, 0.25–2 mm), and microaggregates (MIA, < 0.25 mm) under smooth vetch covering. The relationship between microbial carbon use efficiency and nutrient restriction in soil aggregates was elucidated and the alteration was revealed in microbial metabolic pathways and carbon sequestration functions within soil aggregates under green manure covering. The results showed that: (1) Smooth vetch covering significantly increased the C: N resource imbalance (ln(MBC: MBN)/ln(SOC: TN)) in soil aggregates from 0.50 to 0.69, and thus increased the nitrogen requirement of soil aggregates. However, the N:P resource imbalance (ln(MBN: MBP)/ln(TN: TP)) decreased from 2.25 to 1.78, especially in microaggregates. (2) To mitigate the limitation imposed by nitrogen availability at the aggregate level, microorganisms have ramped up the activity of C- (46.72 %–98.64 %), N- (2.32 %–121.00 %), and P- acquiring enzymes (119.11 %–187.78 %). Nevertheless, the spatial heterogeneity inherent in soil aggregates, characterized by varying particle sizes, has led to a pivotal shift in microbial nutrient limitation. As soil aggregate size diminishes, the primary constraint on microbial activity transitions from nitrogen limitation to phosphorus limitation, reflecting the dynamic interplay between soil structure and nutrient availability. (3) In addition, smooth vetch covering increased the microbial carbon use efficiency of soil aggregates, and it increased with the increase of the particle size of soil aggregates: LMA (178.80 %) > SMA (147.23 %) > MIA (9.99 %). Microorganisms allocated more energy to generate biomass instead of obtaining limiting nutrient elements. These results indicate that smooth vetch covering could increase the organic carbon content of soil aggregates. In addition, in order to adapt to nutrient imbalance, soil aggregates microorganisms choose a “egoistic” metabolic pathway, which alleviates nutrient limitation and assimilated more energy and nutrients into their own biomass and reduces the release of organic carbon. It is evident that integrating Vicia villosa Roth var. glabresens Koch as a ‘green manure’ in orchards serves a dual purpose: it mitigates the environmental impact of chemical fertilizers while simultaneously promoting soil carbon sequestration. This approach offers a robust theoretical framework for achieving the harmonious integration of ecological sustainability and economic viability in economic forestry, such as citrus orchards.

Extracellular enzyme activity and stoichiometry reveal P limitation in the wild panda habitat of the Qinling Mountains

AbstractSoil microbial extracellular enzymes play crucial roles in soil carbon and nutrient cycling by catalyzing soil biochemical processes. However, the activity and stoichiometry of enzymes involved in the carbon, nitrogen, and phosphorus cycles in the environment with the highest density of wild giant pandas on the southern slopes of the Qinling Mountains is unknown. We have established eight research areas at an elevation of 1090–2621 m. The β‐1,4‐glucosidase (BG), β‐1,4‐N‐acetylglucosaminidase (NAG), and acid phosphatase (APase) in soil samples were measured. We used redundancy analysis and structural equation model to evaluate the driving factors of metabolic restriction of soil microorganisms along the elevational gradient, and four models were used to cross‐evaluate the nutrient restriction status of soil microorganisms. The results showed that most soil physiochemical properties, soil microbial biomass, and microbial extracellular enzymes exhibited a hump‐shaped trend with increasing elevation. Elevation indirectly affected soil enzyme activity and stoichiometry by C, N, and P status. Microorganisms are limited by C at lower and higher elevations but limited by N at medium elevations. These results could help strengthen the conservation and management of the wild panda's natural habitat.

Vegetation drives soil microbial metabolic limitation through modifications of soil properties and microbial biomass during desert grassland-shrubland state anthropogenic transition

Shrub encroachment became widespread in grass-dominated ecosystem regions. However, there is still limited attention on the changes in plant-soil system C:N:P stoichiometry and soil microbial metabolic limitation status caused by anthropogenic encroachment of shrubs in the desert grassland of northwest China, as well as their driving mechanisms. In this study, we quantified the C:N:P stoichiometry of plant-litter-soil, soil physicochemical properties, microbial biomass, extracellular enzyme activities, and microbial metabolic limitation status during the anthropogenic transition from grassland to shrubland. Here, our results showed that anthropogenic shrub encroachment reduced plant C, soil water, and soil nutrient contents while increasing N and P contents in both plants and litter. The correlation also suggested that litter nutrient content relied on plant nutrients, while plants depleted soil nutrients. Secondly, with the gradient of shrub encroachment, the stoichiometry (except for soil C:N ratio) of plant-litter-soil showed a decreasing trend and a positive correlation. Furthermore, our study indicated that shrub encroachment reduced soil microbial biomass and extracellular enzyme activities, altering microbial stoichiometry and patterns of enzyme allocation. Redundancy analysis revealed that microbial biomass, extracellular enzyme activities, and stoichiometry were mainly driven by soil water content, nutrient content, and nutrient stoichiometry. The vector-threshold element ratio model revealed that along the transition from desert grassland to shrubland, soil microbial nutrient limitation shifted from P to N, while energy (C) limitation intensified. The soil microbial metabolic limitation was driven jointly by plants and litter through modifications of soil properties and microbial biomass. Additionally, soil properties played a crucial role among these factors. In summary, over the past 40 years, shrub anthropogenic encroachment has formed a desert grassland-shrub mosaic landscape, depleted soil water and nutrient contents, and altered ecological stoichiometry and microbial metabolic limitation patterns in northwest China. This study provides new insights into the C, N, and P cycling in plant-soil systems following shrub encroachment caused by global climate change and anthropogenic disturbances.

Biofilms from the Kapova Cave Walls as a Source of Hydrolase Producers

The studies of bacterial communities from extreme econiches are presently aimed mainly at analyzing the biodiversity of microorganisms using molecular biology methods. Cultivated bacteria from karst caves represent a unique group of microorganisms, the biochemical potential of which has been poorly studied. In the present work, bacteria from biofilms on the walls of the Kapova Cave (Shulgan-Tash Nature Reserve, Bashkortostan) were isolated and characterized in order to assess the ability of identified isolates to produce extracellular hydrolytic enzymes. Most of the isolates (89%) were members of the phylum Proteobacteria, with the remaining ones belonging to the phyla Actinobacteria, Firmicutes, and Bacteroidetes, which accounted for 5, 4, and 2% of the isolates, respectively. Strains with high levels of secreted protease, RNase, and amylase activity were identified as Stenotrophomonas rhizophila, Lysinibacillus fusiformis, and Pseudomonas stutzeri, respectively.

Understanding the differential impact of PFOS and F-53B pollution on reed-mediated soil organic matter decomposition: Insights from rhizosphere priming effect

Plants affect soil microorganisms through the release of root exudates under pollution stress. This process may affect rhizosphere priming effect (RPE) and alter the rate of soil organic matter decomposition. However, the influence of plants on the decomposition of organic matter in soil subjected to pollution stress remains unclear. We studied the effects of exposure to perfluorooctanesulfonic (PFOS) and its alternative, chlorinated polyfluoroalkyl ether sulfonic (F-53B), at concentrations of 0.1 mg/kg and 50 mg/kg on the RPE of reed. We conducted our experiments in an artificial climate chamber and used the natural 13C tracer method to determine RPE. In the PFOS-exposed groups, the RPE was negative, with values of −11.45 mg C kg−1 soil d−1 in the low PFOS group and −8.04 mg C kg−1 soil d−1 in the high PFOS group. In contrast, in the F-53B-exposed groups, the RPE was positive, with values of 8.26 mg C kg−1 soil d−1 in the low F-53B group and 12.18 mg C kg−1 soil d−1 in the high F-53B group. Exposure of reeds to PFOS/F-53B stress resulted in differential effects on extracellular enzyme activities. The observed positive and negative RPE phenomena could be attributed to variations in extracellular enzyme activities. In conclusion, RPE responded differently under PFOS/F-53B exposure.

Anti-Cancer Potential of Isoflavone-Enriched Fraction from Traditional Thai Fermented Soybean against Hela Cervical Cancer Cells.

Cervical cancer is a leading cause of gynecological malignancies and cancer-related deaths among women worldwide. This study investigates the anti-cancer activity of Thua Nao, a Thai fermented soybean, against HeLa cervical carcinoma cells, and explores its underlying mechanisms. Our findings reveal that the ethyl acetate fraction of Thua Nao (TN-EA) exhibits strong anti-cancer potential against HeLa cells. High-performance liquid chromatography (HPLC) analysis identified genistein and daidzein as the major isoflavones in TN-EA responsible for its anti-cancer activity. TN-EA and genistein reduced cell proliferation and induced G2/M phase arrest, while daidzein induced G1 arrest. These responses were associated with the downregulation of cell cycle regulators, including Cyclin B1, cycle 25C (Cdc25C), and phosphorylated cyclin-dependent kinase 1 (CDK-1), and the upregulation of the cell cycle inhibitor p21. Moreover, TN-EA and its active isoflavones promoted apoptosis in HeLa cells through the intrinsic pathway, evidenced by increased levels of cleaved Poly (ADP-ribose) polymerase (PARP) and caspase-3, loss of mitochondrial membrane potential, and the downregulation of anti-apoptotic proteins B-cell leukemia/lymphoma 2 (Bcl-2), B-cell lymphoma-extra-large (Bcl-xL), cellular inhibitor of apoptosis proteins 1 (cIAP), and survivin. Additionally, TN-EA and its active isoflavones effectively reduced cell invasion and migration by downregulating extracellular matrix degradation enzymes, including Membrane type 1-matrix metalloproteinase (MT1-MMP), urokinase-type plasminogen activator (uPA), and urokinase-type plasminogen activator receptor (uPAR), and reduced the levels of the mesenchymal marker N-cadherin. At the molecular level, TN-EA suppressed STAT3 activation via the regulation of JNK and Erk1/2 signaling pathways, leading to reduced proliferation and invasion of HeLa cells.

Imaging Pulmonary Fibrosis and Treatment Efficacy In Vivo with Autotaxin-Specific PET Ligand [18F]ATX-1905.

Idiopathic pulmonary fibrosis (IPF) is a fatal disease characterized by unpredictable progression and limited therapeutic options. Current diagnosis relies on high resolution computed tomography (HRCT), which may not adequately capture early signs of deterioration. The enzyme autotaxin (ATX) emerges as a prominently expressed extracellular secretory enzyme in the lungs of IPF patients. The objective of this study was to evaluate the effectiveness of 18F-labeled ATX-targeted tracer [18F]ATX-1905, in comparison with [18F]FDG, for early fibrosis diagnosis, disease evolution monitoring, and treatment efficacy assessment in bleomycin-induced pulmonary fibrosis (BPF) models. To assess treatment efficacy, mice were treated with two commonly used drugs for IPF, pirfenidone or nintedanib, from Day 9 to Day 23 postbleomycin administration. Lung tissue assessments encompassed inflammation severity via H&E staining, and Ashcroft scoring via Masson staining, alongside quantification of ATX expression through ELISA. Positron emission tomography (PET) imaging employing [18F]FDG and [18F]ATX-1905 tracked disease progression pre- and post-treatment. The extent of pulmonary fibrosis corresponded to changes in ATX expression levels in the BPF mouse model. Notably, [18F]ATX-1905 exhibited elevated uptake in BPF lungs during the progression of the disease, particularly evident at the early stage (Day 9). This uptake was inhibited by an ATX inhibitor, PF-8380, underscoring the specificity of the radiotracer. Conversely, [18F]FDG uptake, peaking at Day 15, decreased subsequently, likely reflective of diminished inflammation. A 2-week treatment regimen using either pirfenidone or nintedanib resulted in notable reductions of ATX expression levels and fibrosis degrees within lung tissues, based on ELISA and Masson staining, as evidenced by PET imaging with [18F]ATX-1905. [18F]FDG uptake also decreased following the treatment period. Additionally, PET/CT imaging extended to a nonhuman primate (NHP) BPF model. The uptake of [18F]ATX-1905 (SUVmax = 2.2) was significantly higher than that of [18F]FDG (SUVmax = 0.7) in fibrotic lung tissue. Using our novel ATX-specific radiotracer [18F]ATX-1905 and PET/CT imaging, we demonstrated excellent ability in early fibrosis detection, disease monitoring, and treatment assessment within lungs of the BPF mouse models. [18F]ATX-1905 displayed remarkable specificity for ATX expression and high sensitivity for ATX alterations, suggesting its potential for monitoring varying ATX expression in lungs of IPF patients.

Isolation and Identification of Swan (Cygnus olor) Digestive Tract Cellulothic Bacteria to Support Fiber Degradation

Cellulolytic bacteria are bacteria that produce cellulase enzymes capable of degrading cellulose substrates. This study aimed to isolate and identify as well as measure the cellulase enzyme activity of cellulolytic bacteria isolated from the digestive tract of swans (Cygnus olor) in Mataram. The bacteria were isolated using de Man Rogosa Sharpe (MRS) selective media with the spread plate method. Then, the extracellular enzyme activity test was conducted by growing selected pure isolates on 1% Carboxy Methyl Cellulose (CMC 1%) media, followed by pouring 0.1% congo red solution and rinsing with 1M NaCl solution to determine cellulolytic activity (cellulolytic potential indicated by the appearance of a clear zone around the colony) and measuring the cellulolytic index. The bacterial isolation results obtained 4 isolates with the potential to support fiber degradation in animal feed. The highest cellulolytic index was produced by the isolate coded S1, reaching 34.5 mm, while the lowest cellulolytic index was produced by 2 isolates coded J21 and I2, reaching 18.8 mm.

Postfire extracellular enzyme activity in a temperate montane forest

AbstractWildfire is a disturbance expected to increase in frequency and severity, changes that may impact carbon (C) dynamics in the soil ecosystem. Fire changes the types of C sources available to soil microbes, increasing pyrogenic C and coarse downed wood, and if there is substantial tree mortality, decreasing C from root exudates and leaf litter. To investigate the impact of this shift in the composition of C resources on microbial processes driving C cycling, we examined microbial activity in soil sampled from an Oregon burn 1 year after fire from sites spanning a range in soil burn severity from unburned to highly burned. We found evidence that postfire rhizosphere priming loss may reduce soil C loss after fire. We measured the potential activity of C‐acquiring and nitrogen (N)‐acquiring extracellular enzymes and contextualized the microbial resource demand using measurements of mineralizable C and N. Subsurface mineralizable C and N were unaltered by fire and negatively correlated with hydrolytic extracellular enzyme activity (EEA) in unburned, but not burned sites. EEA was lower in burned sites by up to 46%, but only at depths below 5 cm, and with greater decreases in sites with high soil burn severity. These results are consistent with a subsurface mechanism driven by tree mortality. We infer that in sites with high tree mortality, subsurface EEAs decreased due to loss of rhizosphere priming and that inputs of dead roots contributed to mineralizable C stabilization.

Characteristics of Rhizosphere Microbiome, Soil Chemical Properties, and Plant Biomass and Nutrients in Citrus reticulata cv. Shatangju Exposed to Increasing Soil Cu Levels.

The prolonged utilization of copper (Cu)-containing fungicides results in Cu accumulation and affects soil ecological health. Thus, a pot experiment was conducted using Citrus reticulata cv. Shatangju with five Cu levels (38, 108, 178, 318, and 388 mg kg-1) to evaluate the impacts of the soil microbial processes, chemistry properties, and citrus growth. These results revealed that, with the soil Cu levels increased, the soil total Cu (TCu), available Cu (ACu), organic matter (SOM), available potassium (AK), and pH increased while the soil available phosphorus (AP) and alkali-hydrolyzable nitrogen (AN) decreased. Moreover, the soil extracellular enzyme activities related to C and P metabolism decreased while the enzymes related to N metabolism increased, and the expression of soil genes involved in C, N, and P cycling was regulated. Moreover, it was observed that tolerant microorganisms (e.g., p_Proteobacteria, p_Actinobacteria, g_Lysobacter, g_Sphingobium, f_Aspergillaceae, and g_Penicillium) were enriched but sensitive taxa (p_Myxococcota) were suppressed in the citrus rhizosphere. The citrus biomass was mainly positively correlated with soil AN and AP; plant N and P were mainly positively correlated with soil AP, AN, and acid phosphatase (ACP); and plant K was mainly negatively related with soil β-glucosidase (βG) and positively related with the soil fungal Shannon index. The dominant bacterial taxa p_Actinobacteriota presented positively correlated with the plant biomass and plant N, P, and K and was negatively correlated with plant Cu. The dominant fungal taxa p_Ascomycota was positively related to plant Cu but negatively with the plant biomass and plant N, P, and K. Notably, arbuscular mycorrhizal fungi (p_Glomeromycota) were positively related with plant P below soil Cu 108 mg kg-1, and pathogenic fungi (p_Mortierellomycota) was negatively correlated with plant K above soil Cu 178 mg kg-1. These findings provided a new perspective on soil microbes and chemistry properties and the healthy development of the citrus industry at increasing soil Cu levels.