Enzyme Pool Research Articles

Mucin-driven ecological interactions in an in vitro synthetic community of human gut microbes.

Specific human gut microbes inhabit the outer mucus layer of the gastrointestinal tract. Certain residents of this niche can degrade the large and complex mucin glycoproteins that constitute this layer and utilise the degradation products for their metabolism. In turn, this microbial mucin degradation drives specific microbiological ecological interactions in the human gut mucus layer. However, the exact nature of these interactions remains unknown. In this study, we designed and studied an in vitro mucin-degrading synthetic community that included mucin O-glycan degraders and cross-feeding microorganisms by monitoring community composition and dynamics through a combination of 16S rRNA gene amplicon sequencing and qPCR, mucin glycan degradation with PGC-LC-MS/MS, production of mucin-degrading enzymes and other proteins through metaproteomics, and metabolite production with HPLC. We demonstrated that specialist and generalist mucin O-glycan degraders stably co-exist and found evidence for cross-feeding relationships. Cross-feeding on the products of mucin degradation by other gut microbes resulted in butyrate production, hydrogenotrophic acetogenesis, sulfate reduction and methanogenesis. Metaproteomics analysis revealed that mucin glycan degraders Akkermansia muciniphila, Bacteroides spp. and Ruminococcus torques together contributed 92% of the total mucin O-glycan degrading enzyme pool of this community. Furthermore, comparative proteomics showed that in response to cultivation in a community compared to monoculture, mucin glycan degraders increased carbohydrate-active enzymes whereas we also found indications for niche differentiation. These results confirm the complexity of mucin-driven microbiological ecological interactions and the intricate role of carbohydrate-active enzymes in the human gut mucus layer.

Biotransformation of Xanthohumol by Entomopathogenic Filamentous Fungi.

Xanthohumol (1) is a major prenylated flavonoid in hops (Humulus lupulus L.) which exhibits a broad spectrum of health-promoting and therapeutic activities, including anti-inflammatory, antioxidant, antimicrobial, and anticancer effects. However, due to its lipophilic nature, it is poorly soluble in water and barely absorbed from the gastrointestinal tract, which greatly limits its therapeutic potential. One method of increasing the solubility of active compounds is their conjugation to polar molecules, such as sugars. Sugar moiety introduced into the flavonoid molecule significantly increases polarity, which results in better water solubility and often leads to greater bioavailability. Entomopathogenic fungi are well known for their ability to catalyze O-glycosylation reactions. Therefore, we investigated the ability of selected entomopathogenic filamentous fungi to biotransform xanthohumol (1). As a result of the experiments, one aglycone (2) and five glycosides (3-7) were obtained. The obtained (2″E)-4″-hydroxyxanthohumol 4'-O-β-D-(4‴-O-methyl)-glucopyranoside (5) has never been described in the literature so far. Interestingly, in addition to the expected glycosylation reactions, the tested fungi also catalyzed chalcone-flavanone cyclization reactions, which demonstrates chalcone isomerase-like activity, an enzyme typically found in plants. All these findings undoubtedly indicate that entomopathogenic filamentous fungi are still an underexploited pool of novel enzymes.

Two pyridoxal phosphate homeostasis proteins are essential for management of the coenzyme pyridoxal 5'-phosphate in Arabidopsis.

Coenzyme management is important for homeostasis of the pool of active metabolic enzymes. The coenzyme pyridoxal 5'-phosphate (PLP) is involved in diverse enzyme reactions including amino acid and hormone metabolism. Regulatory proteins that contribute to PLP homeostasis remain to be explored in plants. Here, we demonstrate the importance of proteins annotated as PLP homeostasis proteins (PLPHPs) for controlling PLP in Arabidopsis (Arabidopsis thaliana). A systematic analysis indicates that while most organisms across kingdoms have a single PLPHP homolog, Angiosperms have two. PLPHPs from Arabidopsis bind PLP and exist as monomers, in contrast to reported PLP-dependent enzymes, which exist as multimers. Disrupting the function of both PLPHP homologs perturbs vitamin B6 (pyridoxine) content, inducing a PLP deficit accompanied by light hypersensitive root growth, unlike PLP biosynthesis mutants. Micrografting studies show that the PLP deficit can be relieved distally between shoots and roots. Chemical treatments probing PLP-dependent reactions, notably those for auxin and ethylene, provide evidence that PLPHPs function in the dynamic management of PLP. Assays in vitro show that Arabidopsis PLPHP can coordinate PLP transfer and withdrawal from other enzymes. This study thus expands our knowledge of vitamin B6 biology and highlights the importance of PLP coenzyme homeostasis in plants.

Specific Features of the Pool of Hydrolytic Enzymes in Soils of Agricultural Terraces in the Eastern Caucasus

Specific Features of the Pool of Hydrolytic Enzymes in Soils of Agricultural Terraces in the Eastern Caucasus

Influence of the Immobilization Technique on the Productivity of Enzymes in the Cascade Reduction of CO2 to CH3OH

The enzymatic effectiveness in the reaction cascade that reduces CO2 to methanol in water at room‐temperature faces various constraints. One of the major challenges is the short life of costly enzymes: immobilization is used to make them more stable and recyclable. The comparative analysis of the several immobilization techniques reported in the literature is challenging due to the diverse reaction conditions (single enzyme test or pool of enzymes test) and experimental setups, as well as the high variability in the amount of enzymes and cofactor. In the present study, a comparison is presented among three different methods (co‐encapsulation into Ca–alginate beads, co‐absorption onto zirconium(IV) phosphate (ZrP) and covalent binding to dialdehydecellulose [DAC]) of co‐immobilization of the three dehydrogenases Fatedehydrogenase (DH), FaldDH, and alcohol dehydrogenase, used in equal amount and under the same experimental conditions, so to check at what extension the support and the immobilization method can influence the activity of the enzymatic pool. DAC is used for the first time to support the three DHs and results to be the best method of immobilization with respect to those used here, that also allows longer life on enzymes and repeated recycling of the supported enzymes, increasing the overall methanol production with respect to the free enzymes.

Transport of enzymes by lymph flow digestive glands

The exosecretes of the digestive glands contain two pools of hydrolytic enzymes - newly formed in the secretory cycle of glandulocytes and recreted from blood flowing to the glands. The second pool of enzymes is repeated many times due to the incretor transport and resorption of hydrolases and zymogens from the secretions of the ductal system of the glands and the chyme of the gastrointestinal tract into the composition of the interstitial fluid and the lymph formed by it. These processes ensure the recycling of digestive gland enzymes, reusable delivery of hydrolases and their inclusion in the composition of exosecretes. The key role in providing enzyme digestion is played by the transport of resorbed and increted enzymes by the lymph flow. This is the digestive role of lymph and lymph flow.

Melatonin in business with abiotic stresses in vegetable crops

Vegetable crops are very important to the agriculture sector since they are the primary source of many of the vitamins and minerals that are required for a balanced diet. In contrast, vegetable production is frequently vulnerable to abiotic stresses such as heavy metals, salinity, cold, heat, and drought stress, which can have a negative impact on yield and quality. Recently, melatonin (MEL) has been identified as a universal plant growth regulator. MEL is a multifunctional, nontoxic, and universal bioactive molecule with low molecular weight. MEL regulate plant growth, protected photosynthetic system, delayed in leaf senescence, promoting root system architecture, maintain ion homeostasis, upregulated antioxidant enzyme pool, increased osmolytes accumulation, and balanced redox homeostasis. MEL also plays an important role in the alleviation of abiotic stress in vegetable crops. Furthermore, MEL restricted heavy metals accumulation and increased mineral nutrient accumulation in many vegetables crops. This study aims to provide a concise overview of the possible impact of MEL on enhancing the resilience of vegetable crop to abiotic stresses.

Presence of key cholinergic enzymes in human spermatozoa and seminal fluid†.

Little is known about the non-neuronal spermic cholinergic system, which may regulate sperm motility and the acrosome reaction initiation process. We investigated the presence of the key acetylcholine (ACh)-biosynthesizing enzyme, choline acetyltransferase (ChAT), and the acetylcholine-degrading enzymes, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) and two ACh-receptors in human spermatozoa and seminal plasma. Fresh ejaculates were used for intra- and extracellular flow cytometric analysis of ChAT, AChE, BChE, and alpha-7-nicotinic and M1-muscarinic ACh-receptors in sperm. For determining the source of soluble enzymes, frozen seminal samples (n = 74) were selected on two bases: (1) from vasectomized (n = 37) and non-vasectomized (n = 37) subjects and (2) based on levels of alpha-glucosidase, fructose, or zinc to define sample subgroups with high or low fluid contribution from the epididymis and seminal vesicle, and prostate, respectively. Flow cytometric analyses revealed that ChAT was expressed intracellularly in essentially all spermatozoa. ChAT was also present in a readily membrane-detachable form at the extracellular membrane of at least 18% of the spermatozoa. These were also highly positive for intra- and extracellular BChE (>83%) and M1 (>84%) and α7 (>59%) ACh-receptors. Intriguingly, the sperm was negative for AChE. Analyses of seminal plasma revealed that spermatozoa and epididymides were major sources of soluble ChAT and BChE, whereas soluble AChE most likely originated from epididymides and seminal vesicles. Prostate had relatively minor contribution to the pool of the soluble enzymes in the seminal fluid. In conclusion, human spermatozoa exhibited a cholinergic phenotype and were one of the major sources of soluble ChAT and BChE in ejaculate. We also provide the first evidence for ChAT as an extracellularly membrane-anchored protein.

Exogenous Putrescine Modulates Nitrate Reductase-Dependent NO Production in Cucumber Seedlings Subjected to Salt Stress.

Polyamines (PAs) are small aliphatic compounds that participate in the plant response to abiotic stresses. They also participate in nitric oxide (NO) production in plants; however, their role in this process remains unknown. Therefore, the study aimed to investigate the role of putrescine (Put) in NO production in the roots of cucumber seedlings subjected to salt stress (120 mM NaCl) for 1 and 24 h. In salinity, exogenous Put can regulate NO levels by managing NO biosynthesis pathways in a time-dependent manner. In cucumber roots exposed to 1 h of salinity, exogenous Put reduced NO level by decreasing nitrate reductase (NR)-dependent NO production and reduced nitric oxide synthase-like (NOS-like) activity. In contrast, during a 24 h salinity exposure, Put treatment boosted NO levels, counteracting the inhibitory effect of salinity on the NR and plasma membrane nitrate reductase (PM-NR) activity in cucumber roots. The role of endogenous Put in salt-induced NO generation was confirmed using Put biosynthesis inhibitors. Furthermore, the application of Put can modulate the NR activity at the genetic and post-translational levels. After 1 h of salt stress, exogenous Put upregulated CsNR1 and CsNR2 expression and downregulated CsNR3 expression. Put also decreased the NR activation state, indicating a reduction in the level of active dephosphorylated NR (dpNR) in the total enzyme pool. Conversely, in the roots of plants subjected to 24 h of salinity, exogenous Put enhanced the NR activation state, indicating an enhancement of the dpNR form in the total NR pool. These changes were accompanied by a modification of endogenous PA content. Application of exogenous Put led to an increase in the amount of Put in the roots and reduced endogenous spermine (Spm) content in cucumber roots under 24 h salinity. The regulatory role of exogenous Put on NO biosynthesis pathways may link with plant mechanisms of response to salt stress.

Functional and biotechnological potential of microbiome associated with soils colonised by cyanobacteria in drylands

Drylands are areas under continuous degradation and desertification frequently covered by cyanobacterial biocrusts. Several studies have indicated that soil microorganisms contribute to nutrient cycling. Nevertheless, little is still known about how those could potentially be participating according to their taxonomic groups. In addition, biocrust-associated communities still remain largely unexplored at taxonomic and functional levels and, therefore, biotechnologically underexploited in comparison with other terrestrial ecosystems. This work aimed at revealing the ecological and biotechnological potential of soil communities using a shotgun sequencing approach in cyanobacterial-colonised biocrusts in the Tabernas Desert regarding the cycles of carbon, nitrogen, phosphorus, iron, sulfur, and potassium. The abundance and successful adaptation of major groups Rubrobacter and Conexibacter was explained by the presence of metabolic diversity regarding glycolytic and CO2 fixation pathways but also by exhibiting a rich pool of hydrolytic enzymes. Besides, Rubrobacter representatives might potentially be driving sulfate mineralisation. Others, such as Caulobacter representatives, were also suggested to have a potential key role in K+ uptake. Furthermore, iron uptake was found to potentially be mediated via synthesis and release of siderophores, mostly derived from pseudomonads. Importantly, this work revealed a meaningful biotechnological niche in which the recovery of novel isolates is a very desirable step forward for the industry and a step forward in unwrapping the potential role of those microorganisms in soil restoration as well as in the functioning of arid soils as CO2 sinks.

Zinc oxide nano-flowers improve the growth and propagation of mulberry cuttings grown under different irrigation regimes by mitigating drought-related complications and enhancing zinc uptake

Silkworm larvae mainly consume mulberry leaves; therefore, mulberry cultivation is important for the production of raw silk. Drought stress and micronutrient deficiency (Zn) are known to affect the propagation of mulberry cuttings. In this purview, the current investigation attempted to inspect the efficacy of different concentrations of zinc oxide nano-flower (ZnNFs) applied through both soil admixture and foliar spray on the propagation of mulberry cuttings grown under deficit irrigation regimes. The overall results demonstrated that the ZnNF-treated plant cuttings were well-adapted to drought stress and performed better in comparison to the control set. Out of the tested concentrations – ZnNF-10 (applied as 10 mg/kg soil and 10 ppm as foliar spray thrice) was found to be optimum, showing relatively better initial root establishment, the emergence of leaves, and survival and sprouting percentage. Further studies also confirmed an improvement in the accumulation of photosynthetic pigments, carbohydrates, and protein content even under extreme drought conditions. Most importantly, the ZnNF-10 treatment contributed to ROS detoxification and cell membrane protection by enhancing the pool of antioxidant enzymes. The study further demonstrated that ZnNF-10 application enhanced zinc content by 147.50%, 179.49%, and 171.99% in root, shoot, and leaves of the treated cuttings; thereby, improving the bioaccumulation factor of the plant parts. All of these interactive phenomena led to an increment in shoot height, biomass, leaf area, and leaf number of cuttings. These findings, therefore, indicated that ZnNFs can be developed as a promising nano-fertilizer for mulberry growth facilitating Zn uptake and mitigation of drought-induced complications.

Soil biochemical inticators of initial presence of fat in different archaeological contexts

The article proposes a new biochemical approach for the reconstruction of the initial presence of fat-containing products in different archaeological contexts (ceramic vessels from burials, soil samples in different parts of the skeleton and cultural layers of archaeological sites) based on the study of qualitative and quantitative changes in the parameters of the soil microbial community, namely, specific groups of microorganisms (lipolytics), a number of lipolytic enzymes, as well as the utilization spectrum of readily available low molecular weight sub-strates. Ground samples of the studied objects were collected in the following regions: ceramic vessels — the Republic of North Ossetia-Alania and the Chechen Republic; burials — Krasnodar Krai; the cultural layer of the settlement — Lipetsk region. The number of lipolytic microorganisms and the level of enzymatic activity in the soil directly depend on the amount of the incoming substrate, in the decomposition of which they participate. After the decomposition of organic residues in the soil, a microbial and enzyme pools are formed, which can persist for a long period. The obtained preliminary data on the study of the decomposition dynamics of fatty substrates give us possibility for the reconstruction of the initial presence of fat in different archaeological contexts using the methods of soil microbiology and biochemistry. But, for a more accurate extrapolation of the results of a model experiment to archaeological objects, more points of observation in time are needed, since the introduction of substrates with different properties and composition can provoke microbial community succession in different ways. Hence, the equilibrium state of the microbial community in each variant of the experiment will be reached at different times. However, the results of the study of soils and cultural layers of archaeological sites of Bronze Age and early me-dieval time have convincingly shown the possibility of applying our approach. As we assumed, the maximum li-pase activity was found in the soil samples under the skull, chest and pelvis, i.e. in areas of human body with the highest content of fat tissues. This showed the possibility for reconstruction the original contents of the vessels from burials using the methods of soil microbiology and biochemistry. A high number of lipolytic microorganisms and lipase activity were detected only in 15–20 % of the vessels. We suggest that fat food may not have been as widely used in the funeral rite as ritual food. The study of lipase activity made it possible to clarify the features of the economic usage of the territory of archaeological site, to identify possible places for cooking.

Thermostable DNA ligases from hyperthermophiles in biotechnology.

DNA ligase is an important enzyme ubiquitous in all three kingdoms of life that can ligate DNA strands, thus playing essential roles in DNA replication, repair and recombination in vivo. In vitro, DNA ligase is also used in biotechnological applications requiring in DNA manipulation, including molecular cloning, mutation detection, DNA assembly, DNA sequencing, and other aspects. Thermophilic and thermostable enzymes from hyperthermophiles that thrive in the high-temperature (above 80°C) environments have provided an important pool of useful enzymes as biotechnological reagents. Similar to other organisms, each hyperthermophile harbors at least one DNA ligase. In this review, we summarize recent progress on structural and biochemical properties of thermostable DNA ligases from hyperthermophiles, focusing on similarities and differences between DNA ligases from hyperthermophilic bacteria and archaea, and between these thermostable DNA ligases and non-thermostable homologs. Additionally, altered thermostable DNA ligases are discussed. Possessing improved fidelity or thermostability compared to the wild-type enzymes, they could be potential DNA ligases for biotechnology in the future. Importantly, we also describe current applications of thermostable DNA ligases from hyperthermophiles in biotechnology.

Structure of Microbial Communities and Biological Activity in Tundra Soils of the Euro-Arctic Region (Rybachy Peninsula, Russia).

The relevance of the Arctic regions' study is rapidly increasing due to the sensitive response of fragile ecosystems to climate change and anthropogenic pressure. The microbiome is an important component that determines the soils' functioning and an indicator of changes occurring in ecosystems. Rybachy Peninsula is the northernmost part of the continental European Russia and is almost completely surrounded by Barents Sea water. For the first time, the microbial communities of the Entic Podzol, Albic Podzol, Rheic Histosol and Folic Histosol as well as anthropogenically disturbed soils (chemical pollution and human impact, growing crops) on the Rybachy Peninsula were characterized using plating and fluorescence microscopy methods, in parallel with the enzymatic activity of soils. The amount and structure of soil microbial biomass, such as the total biomass of fungi and prokaryote, the length and diameter of fungal and actinomycete mycelium, the proportion of spores and mycelium in the fungal biomass, the number of spores and prokaryotic cells, the proportion of small and large fungal spores and their morphology were determined. In the soils of the peninsula, the fungal biomass varied from 0.121 to 0.669 mg/g soil. The biomass of prokaryotes in soils ranged from 9.22 to 55.45 μg/g of soil. Fungi predominated, the proportion of which in the total microbial biomass varied from 78.5 to 97.7%. The number of culturable microfungi ranged from 0.53 to 13.93 × 103 CFU/g in the topsoil horizons, with a maximum in Entic Podzol and Albic Podzol soils and a minimum in anthropogenically disturbed soil. The number of culturable copiotrophic bacteria varied from 41.8 × 103 cells/g in a cryogenic spot to 5551.3 × 103 cells /g in anthropogenically disturbed soils. The number of culturable oligotrophic bacteria ranged from 77.9 to 12,059.6 × 103 cells/g. Changes in natural soils because of anthropogenic impact and a change in vegetation types have led to a change in the structure of the community of soil microorganisms. Investigated tundra soils had high enzymatic activity in native and anthropogenic conditions. The β-glucosidase and urease activity were comparable or even higher than in the soils of more southern natural zone, and the activity of dehydrogenase was 2-5 times lower. Thus, despite the subarctic climatic conditions, local soils have a significant biological activity upon which the productivity of ecosystems largely depends. The soils of the Rybachy Peninsula have a powerful enzyme pool due to the high adaptive potential of soil microorganisms to the extreme conditions of the Arctic, which allows them to perform their functions even under conditions of anthropogenic interference.

Chytrids in Soil Environments: Unique Adaptations and Distributions

Chytridiomycota (zoosporic true fungi) have a consistent presence in soils and have been frequently identified within many diverse terrestrial environments. However, Chytridiomycota and other early-diverging fungi have low representation in whole-genome sequencing databases compared to Dikarya. New molecular techniques have provided insights into the diversity and abundance of chytrids in soils and the changes in their populations both spatially and temporally. Chytrids complete their life cycle within rapidly changing soil environments where they may be more common within micropores due to protection from predation, desiccation, and extreme temperatures. Reproductive and morphological changes occur in response to environmental changes including pH, fluctuating nutrient concentrations, and metals at levels above toxic thresholds. Rhizoids share some features of hyphae, including the spatial regulation of branching and the ability to attach, adapt to, and proliferate in different substrates, albeit on a microscale. Soil chytrids provide a pool of novel enzymes and proteins which enable a range of lifestyles as saprotrophs or parasites, but also can be utilised as alternative tools with some biotechnological applications. Thus, 3D live-cell imaging and micromodels such as MicroCT may provide insight into zoospore functions and rhizoid plasticity, respectively, in response to various conditions. A combination of classical techniques of soil chytrid baiting with simultaneous molecular and ecological data will provide insights into temporal population changes in response to environmental change. The authors emphasise the need to review and improve DNA-based methodologies for identifying and quantifying chytrids within the soil microbiome to expand our knowledge of their taxonomy, abundance, diversity, and functionality within soil environments.

Ecotoxicity of soil Pb pollution reflected by soil β-glucosidase: Comparison of extracellular and intracellular enzyme pool

Lead (Pb) is a major environmental pollutant that threatens the soil environment and human health. Monitoring and assessing Pb toxicity on soil health are of paramount importance to the public. To use soil enzymes as biological indicators of Pb contamination, herein, the responses of soil β-glucosidase (BG) in different pools of soil (total, intracellular and extracellular enzyme) to Pb contamination were investigated. The results indicated that the intra-BG (intracellular BG) and extra-BG (extracellular BG) responded differently to Pb contamination. While the addition of Pb caused a significant inhibition of the intra-BG activities, the extra-BG activities were only slightly inhibited. Pb showed a non-competitive inhibition to extra-BG, while both non-competitive and uncompetitive inhibition were observed for intra-BG in the tested soils. The dose-response modeling was used to calculate ecological dose ED10, which represents the concentration of Pb pollutant that causes a 10 % reduction in Vmax, to express the ecological consequences of Pb contamination. A positive correlation was found between ecological dose ED10 values of intra-BG and soil total nitrogen (p < 0.05), which suggests soil properties may influence Pb toxicity to soil BG. Based on the differences in ED10 and inhibition rate among different enzyme pools, this study suggests that the intra-BG is more sensitive for Pb contamination assessment. From this, we propose that intra-BG should be considered when evaluating Pb contamination using soil enzymes as indicators.

The bioherbicidal potential of isolated fungi cultivated in microalgal biomass.

This study evaluated the bioherbicidal potential of wild fungi grown on microalgal biomass from the digestate treatment of biogas production. Four fungal isolates were used and the extracts were evaluated for the activity of different enzymes and characterized by gas chromatography coupled with mass spectrometry. The bioherbicidal activity was assessed by application on Cucumis sativus, and the leaf damage was visually estimated. The microorganisms showed potential as agents producing an enzyme pool. The obtained fungal extracts presented different organic compounds, most acids, and when applied to Cucumis sativus, showed high levels of leaf damage (80-100 ± 3.00%, deviation relative to the observed average damage). Therefore, the microbial strains are potential biological control agents of weeds, which, together with the microalgae biomass, offer the appropriate conditions to obtain an enzyme pool of biotechnological relevance and with favorable characteristics to be explored as bioherbicides, addressing aspects within the environmental sustainability.

A novel gene therapy for methamphetamine- induced cognitive disorder with a hyper-acidified fusion variant of DnaJB1

Methamphetamine (MA) is spread worldwide and is a highly addictive psychostimulant that can induce neurodegeneration and cognitive disorder, which lacks effective treatments. We and other researchers have found that the crucial member of Hsp70 chaperone machinery, DnaJ, is liable to be co-aggregated with aberrant proteins, which has been confirmed a risk factor to promote neurodegeneration. In the current study, we demonstrated that tailing with a hyper-acidic fusion partner, tua2, human DnaJB1 could resist the formation of toxic mutant Tau aggregates both in prokaryote and eukaryote models. We found that aberrant Tau aggregates could deplete the antioxidant enzyme pool and disturb Hsp70 molecular chaperone system by co-aggregating with the principal members of these systems. Stability-enhanced DnaJB1-tua2 could stop the chain reaction of Tau aggregates as well as maintain redox balance and protein homeostasis. With an MA-induced cognitive disorder mouse model, we found that the cognitive disorder of MA mice was rescued and the overactivated inflammatory response was relieved by the expression of DnaJB1-tua2 in the hippocampus. Furthermore, the Tau neurofibrillary tangles and apoptotic neurons were diminished with the escorting of DnaJB1-tua2. These findings demonstrate that delivering DnaJB1-tua2 in hippocampus may have a therapeutic potential in the treatment of MA-induced cognitive disorder.

Inhibition of Extracellular Enzyme Activity by Reactive Oxygen Species upon Oxygenation of Reduced Iron-Bearing Minerals.

The dual roles of minerals in inhibiting and prolonging extracellular enzyme activity in soils and sediments are governed by enzyme adsorption to mineral surfaces. Oxygenation of mineral-bound Fe(II) generates reactive oxygen species (ROS), yet it is unknown whether and how this process alters the activity and functional lifespan of extracellular enzymes. Here, the effect of mineral-bound Fe(II) oxidation on the hydrolytic activity of a cellulose-degrading enzyme β-glucosidase (BG) was studied using two pre-reduced Fe-bearing clay minerals (nontronite and montmorillonite) and one pre-reduced iron oxide (magnetite) at pH 5 and 7. Under anoxic conditions, BG adsorption to mineral surfaces decreased its activity but prolonged its lifespan. Under oxic conditions, ROS was produced, with the amount of •OH, the most abundant ROS, being positively correlated with the extent of structural Fe(II) oxidation in reduced minerals. •OH decreased BG activity and shortened its lifespan via conformational change and structural decomposition of BG. These results suggest that under oxic conditions, the ROS-induced inhibitory role of Fe(II)-bearing minerals outweighed their adsorption-induced protective role in controlling enzyme activity. These results disclose a previously unknown mechanism of extracellular enzyme inactivation, which have pivotal implications for predicting the active enzyme pool in redox-oscillating environments.

Dissipation kinetics of chlorpyrifos and 3,5,6 trichloro-2-pyridinol under vegetation of different aromatic grasses: Linkage with enzyme kinetics and microbial community of soil

The dissipation of chlorpyrifos (CP) and its hydrolytic metabolite 3,5,6-trichloro-2-pyridinol (TCP) in the soil is crucial for safe agriculture. However, there is still lacking relevant information about its dissipation under different vegetation for remediation purposes. In the present study, evaluation of dissipation of CP and TCP in non-planted and planted soil with different cultivars of three types of aromatic grass viz Cymbopogon martinii (Roxb. Wats), Cymbopogon flexuosus, and Chrysopogon zizaniodes (L.) Nash was examined in light of soil enzyme kinetics, microbial communities, and root exudation. Results revealed that the dissipation of CP was well-fitted into a single first-order exponential model (SFO). A significant reduction in the half-life (DT50) of CP was observed in planted soil (30–63 days) than in non-planted soil (95 days). The presence of TCP in all soil samples was observed. The three types of the inhibitory effect of CP i.e. linear mixed inhibition (increase in enzyme-substrate affinity (Km) and decrease in enzyme pool (Vmax), un-competitive inhibition (decrease in Km and Vmax), and simple competitive inhibition were observed on soil enzymes involved in mineralization of carbon, nitrogen, phosphorus, and sulfur. The improvement in the enzyme pool (Vmax) was observed in planted soil. Streptomyces, Clostridium, Kaistobacter, Planctomyces, and Bacillus were the dominant genera in CP stress soil. CP contamination in soil demonstrated a reduction of richness in microbial diversity and enhancement of functional gene family related to cellular process, metabolism, genetic, and environmental information processing. Among all the cultivars, C. flexuosus cultivars demonstrated a higher dissipation rate of CP along with more root exudation.