Activation Of Enzyme Research Articles

Metabolomics and microbiomics revealed the combined effects of different-sized polystyrene microplastics and imidacloprid on earthworm intestinal health and function

The coexistence of pesticides and plastic film residues in agricultural soils poses a significant threat to soil organisms due to their potential long-term contamination and combined toxic effects. Specifically, earthworms are at risk of simultaneously ingesting residual pesticides and microplastics, yet the impact of this combined exposure on their intestinal health and function remains poorly understood. In this study, earthworm (Eisenia fetida) were single and combined exposed to three particle sizes (10 μm, 500 μm, and 2 mm) of polyethylene microplastics (PE MPs) and imidacloprid (IMI) for 28 days, respectively. Our findings underscore that compared to single exposures, the combined exposure inflicted more profound injuries on intestinal tissues and elicited a heightened activation of intestinal digestive enzymes. Furthermore, the combined exposure significantly perturbed the relative abundance of several pivotal metabolic-associated gut microbiota, fostering an enrichment of pathogenic species. Metabolomics analysis showed combined exposure increased differential metabolites, disrupting amino acid, fatty acid, and carbohydrate metabolism in earthworm intestines, potentially hindering nutrient absorption and causing toxic metabolite accumulation. An integrated omics analysis implies that combined exposures have the potential to disrupt the relative abundance of crucial gut microbiota in earthworms, thereby altering their intestinal metabolism and subsequently impacting intestinal health and functionality. Overall, the results reveal that combined exposure of IMI and PE MPs exacerbate the negative effects on earthworm gut health, and this study holds significant implications for the holistic understanding of the combined toxic effects of microplastics and pesticide on soil ecosystems.

The Effect of Telomerase Enzyme on Cell Aging and Cancer Formation-Treatment

Research has made the effects of the telomerase enzyme on both cellular ageing and cancer even more important. Measurement of telomerase activity appears to be a unique marker in addition to known methods for cancer diagnosis and treatment. In addition, numerous studies have shown that the reduction of telomeres shortens the life span of living cells. It is a known fact that cells with shortened lifespan age earlier and predispose to many diseases, especially cancer types. The fact that old cells are the basis for many diseases and young cells are resistant to many diseases has revealed the reality of keeping the cells young in order to prevent diseases. Studies aimed at understanding the activation of the telomerase enzyme on telomeres, have shown that this enzyme will have a positive effect on cellular rejuvenation on living cells, prolong the life span of living cells, thus preventing age-related diseases before they occur, and also making it impossible for many types of cancer that may develop due to old age to occur in the first place. Predictions such as preventing and stopping cellular senescence in human cells, providing cellular rejuvenation by reversing cellular senescence and extending human life span as much as desired are also worthy of further emphasis with the studies carried out.

Otilonium bromide ameliorates pulmonary fibrosis in mice through activating phosphatase PPM1A.

Pulmonary fibrosis (PF) is a chronic, progressive and irreversible interstitial lung disease characterized by unremitting pulmonary myofibroblasts activation, extracellular matrix (ECM) deposition and inflammatory recruitment. PF has no curable medication yet. In this study we investigated the molecular pathogenesis and potential therapeutic targets of PF and discovered drug lead compounds for PF therapy. A murine PF model was established in mice by intratracheal instillation of bleomycin (BLM, 5 mg/kg). We showed that the protein level of pulmonary protein phosphatase magnesium-dependent 1A (PPM1A, also known as PP2Cα) was significantly downregulated in PF patients and BLM-induced PF mice. We demonstrated that TRIM47 promoted ubiquitination and decreased PPM1A protein in PF progression. By screening the lab in-house compound library, we discovered otilonium bromide (OB, clinically used for treating irritable bowel syndrome) as a PPM1A enzymatic activator with an EC50 value of 4.23 μM. Treatment with OB (2.5, 5 mg·kg-1·d-1, i.p., for 20 days) significantly ameliorated PF-like pathology in mice. We constructed PF mice with PPM1A-specific knockdown in the lung tissues, and determined that by targeting PPM1A, OB treatment suppressed ECM deposition through TGF-β/SMAD3 pathway in fibroblasts, repressed inflammatory responses through NF-κB/NLRP3 pathway in alveolar epithelial cells, and blunted the crosstalk between inflammation in alveolar epithelial cells and ECM deposition in fibroblasts. Together, our results demonstrate that pulmonary PPM1A activation is a promising therapeutic strategy for PF and highlighted the potential of OB in the treatment of the disease.

Recombinant C-Terminal Catalytic Domain of Rat L-Gulono Lactone Oxidase Produced in Bacterial Cells Is Enzymatically Active.

The L-gulonolactone oxidase enzyme (GULO) catalyzes the last step of L-ascorbic acid (vitamin C) biosynthesis. This enzymatic activity is lost in primates. The full-length rat GULO has been previously produced in plants and demonstrated to be active. In this study, we compared the activity of two variants of GULO produced in Escheriachia coli cells, full-length rat GULO (fGULO) and its C-terminal catalytic domain (cGULO). The expression and purification of the recombinant proteins were optimized, and their biological activity was confirmed by two methods, the GULO activity assay in the protein extracts and the 'in-gel' staining for GULO activity. Both variants of recombinant GULO were biologically active in both assays. However, cGULO is more promising than fGULO for ascorbic acid production because it is more efficiently produced by bacteria. Furthermore, the optimal activities of the fGULO and cGULO recombinant proteins were observed at pH 7 and 6.5, and at temperatures of 40 and 30 °C, respectively. Kinetic studies revealed that at low substrate concentrations, Km values for fGULO and cGULO were 53.5 ± 5 and 42 ± 6.3 µM, respectively.

Development of Brain Penetrant Pyridazine Pantothenate Kinase Activators.

Conversion of pantothenate to phosphopantothenate in humans is the first dedicated step in the coenzyme A (CoA) biosynthesis pathway and is mediated by four isoforms of pantothenate kinase. These enzymes are allosterically regulated by acyl-CoA levels, which control the rate of CoA biosynthesis. Small molecule activators of the PANK enzymes that overcome feedback suppression increase CoA levels in cultured cells and animals and have shown great potential for the treatment of pantothenate kinase-associated neurodegeneration and propionic acidemias. In this study, we detail the further optimization of PANK pyridazine activators using structure-guided design and focus on the cellular CoA activation potential, metabolic stability, and solubility as the primary drivers of the structure-activity relationship. These studies led to the prioritization of three late-stage preclinical lead PANK modulators with improved pharmacokinetic profiles and the ability to substantially increase brain CoA levels. Compound 22 (BBP-671) eventually advanced into clinical testing for the treatment of PKAN and propionic acidemia.

Micronutrient deficiency-induced oxidative stress in plants.

Micronutrients like iron (Fe), zinc (Zn), copper (Cu), manganese (Mn), boron (B), nickel (Ni), and molybdenum (Mo) perform significant roles in the regulation of plant metabolism, growth, and development. Micronutrients, namely Fe, Zn, Cu, Mn, and Ni, are involved in oxidative stress and antioxidant defense as they are cofactors or activators of various antioxidant enzymes, viz., superoxide dismutase (Fe, Cu/Zn, Mn, and Ni), catalase (Fe), and ascorbate peroxidase (Fe). An effort has been made to incorporate recent advances along with classical work done on the micronutrient deficiency-induced oxidative stress and associated antioxidant responses of plants. Deficiency of a micronutrient produces ROS in the cellular compartments. Enzymatic and non-enzymatic antioxidant defense systems are often modulated by micronutrient deficiency to regulate redox balance and scavenge deleterious ROS for the safety of cellular constituents. ROS can strike cellular constituents such as lipids, proteins, and nucleic acids and can destruct cellular membranes and proteins. ROS might act as a signaling molecule and activate the antioxidant proteins by interacting with signaling partners such as respiratory burst oxidase homolog (RBOH), G-proteins, Ca2+, mitogen activated protein kinases (MAPKs), and various transcription factors (TFs). Opinions on probable ROS signaling under micronutrient deficiency have been described in this review. However, further research is required to decipher micronutrient deficiency-induced ROS generation, perception, and associated downstream signaling events, leading to the development of antioxidant responses in plants.

Alteration of stress-physiological mechanisms in sRNA-treated sweet corn plants during MDMV infection.

Maize dwarf mosaic virus (MDMV) can significantly reduce the growth and development of susceptible varieties of sweet corn. The virus utilises the energy and reserve sources of plant cells to ensure its reproduction in the microspaces formed by cell membranes. Therefore, the severity of stress can be monitored by examining certain physiological changes, for example, changes in the degree of membrane damage caused by lipid peroxidation, as well as changes in the amount of photosynthetic pigments. The activation of antioxidant enzymes (e.g. ascorbate peroxidase, guaiacol peroxidase, glutathione reductase) and the accumulation of phenolic compounds with antioxidant properties can indirectly protect against the oxidative stress caused by the presence of the positive orientation, single-stranded RNA-virus. This study demonstrates the changes in these physiological processes in a sweet corn hybrid (Zea mays cv. saccharata var. Honey Koern.) susceptible to MDMV infection, and suggests that exogenous small RNA treatment can mitigate the damage caused by virus infection.

Investigation of laccase activity in cholinium-based ionic liquids using experimental and molecular dynamics techniques

Laccases hold great potential for biotechnological applications, particularly in environmental pollutant remediation. Laccase activity is governed by the solvent environment, and ionic liquids (ILs) emerge as a versatile solvent for activation or stabilization of enzymes. Herein, effects of cholinium-based ILs formulated with carboxylic acids, inorganic acid, and amino acids as anionic species, on the catalytic activity of laccase from Trametes versicolor were investigated by experimental and computational approaches. Experimental results showed that laccase activity was enhanced by 21.39 % in 0.5 M cholinium dihydrogen citrate ([Cho][DHC]), in relation to the laccase activity in phosphate buffer medium. However, cholinium aminoate ILs negatively affected laccase activity, as evidenced by the partial deactivation of laccase in both cholinium glycinate and cholinium phenylalaninate, at concentrations of 0.1 M and 0.5 M, respectively. Molecular dynamics studies revealed that the enhancement of laccase activity in [Cho][DHC] might be attributed to the highly stabilized and compact structure of laccase, facilitating a better internal electron transfer during the laccase-substrate interactions. Enhanced catalytic performance of laccase in [Cho][DHC] was postulated to be driven by the high accumulation level of dihydrogen citrate anions around laccase's surface. [Cho][DHC] holds great promise as a cosolvent in laccase-catalyzed biochemical reactions.

Functional loss of ERBB receptor feedback inhibitor 1 (MIG6) promotes glioblastoma tumorigenesis by aberrantactivation of epidermal growth factor receptor (EGFR).

Dysregulation of epidermal growth factor receptor (EGFR) is one of the most common mechanisms associated with the pathogenesis of various cancers. Mitogen-inducible gene 6 [MIG6; also known as ERBB receptor feedback inhibitor 1 (ERRFI1)], identified as a feedback inhibitor of EGFR, negatively regulates EGFR by directly inhibiting its kinase activity and facilitating its internalization, subsequently leading to degradation. Despite its proposed role as an EGFR-dependent tumor suppressor, the functional consequences and clinical relevance in cancer etiology remain incompletely understood. Here, we identify that the stoichiometric balance between MIG6 and EGFR is crucial in promoting EGFR-dependent oncogenic growth in various experimental model systems. In addition, a subset of ERRFI1 (the official gene symbol of MIG6) mutations exhibit impaired ability to suppress the enzymatic activation of EGFR at multiple levels. In summary, our data suggest that decreased or loss of MIG6 activity can lead to abnormal activation of EGFR, potentially contributing to cellular transformation. We propose that the mutation status of ERRFI1 and the expression levels of MIG6 can serve as additional biomarkers for guiding EGFR-targeted cancer therapies, including glioblastoma.

Magnesium induced structural reorganization in the active site of adenylate kinase.

Phosphoryl transfer is a fundamental reaction in cellular signaling and metabolism that requires Mg2+ as an essential cofactor. While the primary function of Mg2+ is electrostatic activation of substrates, such as ATP, the full spectrum of catalytic mechanisms exerted by Mg2+ is not known. In this study, we integrate structural biology methods, molecular dynamic (MD) simulations, phylogeny, and enzymology assays to provide molecular insights into Mg2+-dependent structural reorganization in the active site of the metabolic enzyme adenylate kinase. Our results demonstrate that Mg2+ induces a conformational rearrangement of the substrates (ATP and ADP), resulting in a 30° adjustment of the angle essential for reversible phosphoryl transfer, thereby optimizing it for catalysis. MD simulations revealed transitions between conformational substates that link the fluctuation of the angle to large-scale enzyme dynamics. The findings contribute detailed insight into Mg2+ activation of enzymes and may be relevant for reversible and irreversible phosphoryl transfer reactions.

Germination Promotes Flavonoid Accumulation of Finger Millet (Eleusine coracana L.): Response Surface Optimization and Investigation of Accumulation Mechanism.

Germination is an effective measure to regulate the accumulation of secondary metabolites in plants. In this study, we optimized the germination conditions of finger millet by response surface methodology. Meanwhile, physiological characteristics and gene expression were measured to investigate the mechanism of flavonoid accumulation in finger millet at the germination stage. The results showed that when germination time was 5.7 d, germination temperature was 31.2 °C, and light duration was 17.5 h, the flavonoid content of millet sprouts was the highest (7.0 μg/sprout). The activities and relative gene expression of key enzymes for flavonoid synthesis (phenylalanine ammonia-lyase, 4-coumarate-coenzyme a ligase, and cinnamate 4-hydroxylase) were significantly higher in finger millet sprouts germinated at 3 and 5 d compared with that in ungerminated seeds (p < 0.05). In addition, germination enhanced the activities of four antioxidant enzymes (catalase, peroxidase, superoxide dismutase, and ascorbate peroxidase) and up-regulated the gene expression of PAL and APX. Germination increased malondialdehyde content in sprouts, which resulted in cell damage. Subsequently, the antioxidant capacity of the sprouts was enhanced through the activation of antioxidant enzymes and the up-regulation of their gene expression, as well as the synthesis of active substances, including flavonoids, total phenolics, and anthocyanins. This process served to alleviate germination-induced cellular injury. These findings provide a research basis for the regulation of finger millet germination and the enhancement of its nutritional and functional properties.

The Secreted Aminopeptidase of Pseudomonas aeruginosa (PaAP).

Pseudomonas aeruginosa is an opportunistic pathogen that causes severe infections in compromised hosts. P. aeruginosa infections are difficult to treat because of the inherent ability of the bacteria to develop antibiotic resistance, secrete a variety of virulence factors, and form biofilms. The secreted aminopeptidase (PaAP) is an emerging virulence factor, key in providing essential low molecular weight nutrients and a cardinal modulator of biofilm development. PaAP is therefore a new potential target for therapy of P. aeruginosa infections. The present review summarizes the current knowledge of PaAP, with special emphasis on its biochemical and enzymatic properties, activation mechanism, biological roles, regulation, and structure. Recently developed specific inhibitors and their potential as adjuncts in the treatment of P. aeruginosa infections are also described.

The influence of HSP inducers on salinity stress in sterlet sturgeon (Acipenser ruthenus): In vitro study on HSP expression, immune responses, and antioxidant capacity.

Heat shock proteins (HSPs) play a crucial role in antioxidant systems, immune responses, and enzyme activation during stress conditions. Salinity changes can cause stress and energy expenditure in fish, resulting in mortality, especially in fingerlings. The purpose of this study was to examine the relationship between salinity and HSPs in stressed fish by assessing the effects of various HSP inducers (HSPis), including Pro-Tex® (800mM), amygdalin (80mM), and a novel synthetic compound derived from pirano piranazole (80µM), on isolated cells from Sterlet Sturgeon (Acipenser ruthenus) exposed to 13‰ salinity (S13). After liver, kidney, and gill cells were cultured, the HSPi compounds were treated in vitro in the presence and absence of salinity. The expression patterns of HSP27, HSP70, and HSP90 were assessed by Western blotting. Biochemical enzymes (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and lactate dehydrogenase), cortisol levels, and immune parameters (component 3, immunoglobulin M, and lysozyme) were measured before and after treatment with HSPis and HSPi+S13. According to these findings, HSPis positively modulate HSP expression, immune responses, and antioxidant levels. Furthermore, they increased in vitro cell survival by maintaining cortisol levels and biochemical enzyme activities in A. ruthenus under saline conditions (P<0.0001). In conclusion, HSPis can increase A. ruthenus resistance to salinity stress. However, the results also indicated that these compounds can reverse the adverse effects of salinity. The effectiveness of this approach depends on further research into the effects of these ecological factors on the health status of the species, especially in vivo and in combination with other stresses.

Correlation Between BMI and Severity of Acute Pancreatitis: A Retrospective Study.

Background Pancreatitis, marked by sterile inflammation of the pancreas, can present as either acute or chronic. It involves the premature activation of proteolytic enzymes, leading to autodigestion, inflammation, and potential systemic effects. This study investigates the impact of obesity on the severity of acute pancreatitis, given its role in systemic inflammation and its association with severe morbidity and mortality. Methods A retrospective analysis was conducted on patients treated for acute pancreatitis over a five-month period at the Royal Albert Edward Infirmary, Wrightington, Wigan and Leigh (WLL) NHS Foundation Trust, Wigan, United Kingdom. Patients were evaluated using diagnostic criteria such as abdominal pain, elevated serum enzyme levels, and imaging results. The study explored correlations between BMI and pancreatitis severity, hospital length of stay, and complications, applying the Atlanta severity classification. Results The analysis revealed a weak, statistically insignificant correlation between BMI and the severity of acute pancreatitis, hospital stay length, and complications. This was consistent across various statistical methods, including Pearson correlation coefficients and multiple linear regression. These findings suggest that, while obesity may influence the inflammatory response in acute pancreatitis, it does not have a significant impact on clinical outcomes within this cohort. Conclusions The study highlights the complex role of obesity in exacerbating pancreatic inflammation but also emphasizes the need for larger, more definitive studies to explore this relationship further. It underscores the importance of early recognition and intervention in managing acute pancreatitis, regardless of BMI status.

Microbial carbohydrate active enzyme (CAZyme) genes and diversity from Menagesha Suba natural forest soils of Ethiopia as revealed by shotgun metagenomic sequencing

BackgroundThe global over-reliance on non-renewable fossil fuels has led to the emission of greenhouse gases, creating a critical global environmental challenge. There is an urgent need for alternative solutions like biofuels. Advanced biofuel is a renewable sustainable energy generated from lignocellulosic plant materials, which can significantly contribute to mitigating CO2 emissions. Microbial Carbohydrate Active Enzymes (CAZymes) are the most crucial enzymes for the generation of sustainable biofuel energy. The present study designed shotgun metagenomics approaches to assemble, predict, and annotate, aiming to gain an insight into the taxonomic diversity, annotate CAZymes, and identify carbohydrate hydrolyzing CAZymes from microbiomes in Menagesha suba forest soil for the first time.ResultsThe microbial diversity based on small subunit (SSU) rRNA analysis revealed the dominance of the bacterial domain representing 81.82% and 92.31% in the studied samples. Furthermore, the phylum composition result indicated the dominance of the phyla Proteobacteria (23.08%, 27.27%), Actinobacteria (11.36%, 20.51%), and Acidobacteria (10.26%, 15.91%). The study also identified unassigned bacteria which might have a unique potential for biopolymer hydrolysis. The metagenomic study revealed that 100,244 and 65,356 genes were predicted from the two distinct samples. A total number of 1806 CAZyme genes were identified, among annotated CAZymes, 758 had a known enzyme assigned to CAZymes. Glycoside hydrolases (GHs) CAZyme family contained most of the CAZyme genes with known enzymes such as β-glucosidase, endo-β-1,4-mannanase, exo-β-1,4-glucanase, α-L-arabinofuranosidase and oligoxyloglucan reducing end-specific cellobiohydrolase. On the other hand, 1048 of the identified CAZyme genes were putative CAZyme genes with unknown enzymatical activity and the majority of which belong to the GHs family.ConclusionsIn general, the identified putative CAZymes genes open up an opportunity for the discovery of new enzymes responsible for hydrolyzing biopolymers utilized for biofuel energy generation. This finding is used as a first-hand piece of evidence to serve as a benchmark for further and comprehensive studies to unveil novel classes of bio-economically valuable genes and their encoded products.

Effect of Zinc on Germination, Growth Yield of (Solanum melongena L.)

Eggplant (Solanum melongena L.) is an economically significant crop, valued for its culinary versatility and nutritional content. Understanding the factors that influence its growth is crucial for optimizing agricultural practices and improving yield. Zinc, an essential micronutrient, plays a vital role in various physiological processes within plants, including enzyme activation, photosynthesis, and hormone regulation. This study investigated the effect of zinc supplementation on the growth parameters of eggplant. The experiment was conducted in Botany Lab of Maharishi University of Information Technology, Lucknow in earthen Pots filled with garden soil. Where eggplant seedlings were subjected to different levels of zinc supplementation. Parameters such as plant height, leaf area, root development, and biomass accumulation were measured at regular intervals over the growth period. Results indicated a significant positive correlation between zinc concentration and various growth parameters of eggplant. Seedlings treated with higher zinc concentrations, enhanced leaf expansion, and more extensive root systems compared to those with lower or no zinc supplementation. Furthermore, zinc-treated eggplants displayed improved resistance to certain environmental stressors, suggesting a potential role of zinc in enhancing plant resilience. These findings underscore the importance of adequate zinc supply in promoting the growth and development of eggplant crops, thereby contributing to higher yields and improved agricultural sustainability. Further research is warranted to elucidate the underlying mechanisms of zinc-mediated growth enhancement in eggplant and optimize zinc application strategies for maximum benefit.

Increased Expression of Mevalonate Pathway-Related Enzymes in Angiotensin II-Induced Abdominal Aortic Aneurysms.

Abdominal aortic aneurysm (AAA) is characterized by permanent luminal expansion and a high mortality rate due to aortic rupture. Despite the identification of abnormalities in the mevalonate pathway (MVA) in many diseases, including cardiovascular diseases, the potential impact of this pathway on AAA remains unclear. This study aims to investigate whether the expression of the MVA-related enzyme is altered during the progression of angiotensin II (Ang II) -induced AAA.Ang II 28D and Ang II 5D groups were continuously perfused with Ang II for 28 days and 5 days, respectively, and the Sham group was perfused with saline. The general and remodeling characteristics of AAA were determined by biochemical and histological analysis. Alteration of MVA-related enzyme expressions was revealed by western blot and single-cell RNA sequencing (scRNA-seq).The continuous Ang II infusion for 28 days showed significant aorta expansion and arterial remodeling. Although the arterial diameter slightly increased, the aneurysm formation was not found in Ang II induction for 5 days. MVA-related enzyme expression and activation of small GTP-binding proteins were significantly increased after Ang II-induced. As verified by scRNA-seq, the key enzyme gene expression was also higher in Ang II 28D. Similarly, it was detected that the expression levels of the above enzymes and the activity of small G proteins were elevated in the early stage of AAA as induced by Ang II infusion for 5 days.Continuous Ang II infusion-induced abdominal aortic expansion and arterial remodeling were accompanied by altered expression of key enzymes in the MVA.

Coupling sensor to enzyme in the voltage sensing phosphatase

Voltage-sensing phosphatases (VSPs) dephosphorylate phosphoinositide (PIP) signaling lipids in response to membrane depolarization. VSPs possess an S4-containing voltage sensor domain (VSD), resembling that of voltage-gated cation channels, and a lipid phosphatase domain (PD). The mechanism by which voltage turns on enzyme activity is unclear. Structural analysis and modeling suggest several sites of VSD-PD interaction that could couple voltage sensing to catalysis. Voltage clamp fluorometry reveals voltage-driven rearrangements in three sites implicated earlier in enzyme activation—the VSD-PD linker, gating loop and R loop—as well as the N-terminal domain, which has not yet been explored. N-terminus mutations perturb both rearrangements in the other segments and enzyme activity. Our results provide a model for a dynamic assembly by which S4 controls the catalytic site.

The effect of Arbuscular Mycorrhizal fungal species Funneliformis mosseae and biochar against Verticillium dahliae in pepper plants under salt stress

Both biotic and abiotic stress factors play an important role in reducing the growth and productivity of many crops. In this study, the effects of arbuscular mycorrhizal (AM) fungi and biochar (Bc) were investigated against Verticillium dahliae (Vd) disease in pepper plants grown under salt stress. These effects were evaluated on enzyme activities, salt tolerance, disease severity, plant growth and physiological characteristics. In this study, pepper was treated with 2% biochar, Funneliformis mosseae (Fm), Vd and different concentrations of salt (0 mM, 50 mM, 100 mM, 150 mM). The interaction of Fm and 2% Bc significantly increased the morphological parameters of the plant and the plant tolerated salt and Vd under combined high stress conditions of biotic (Vd) and abiotic (salt) stresses. It was also shown that Fm had a different effect than Bc on the membrane injury index (MII; %) and had a significant effect on leaf relative water content (LRWC; %). In addition, Vd increased mycorrhizal reliance, with the higher spore density of AMF higher disease severity observed in treatments involving Fm and Fm + Bc. Furthermore, phenol and antioxidant values were altered in Vd treatments, while Fm decreased CAT enzyme activation. Therefore, this study supports that AMF + biochar used in sustainable agriculture increases plant resistance to the soil pathogen (V. dahliae) and the salt stresses.

Corynebacterium glutamicum pyruvate:quinone oxidoreductase: an enigmatic metabolic enzyme with unusual structural features.

Pyruvate:quinone oxidoreductase (PQO) is a flavin-containing peripheral membrane enzyme catalyzing the decarboxylation of pyruvate to acetate and CO2 with quinone as an electron acceptor. Here, we investigate PQO activity in Corynebacterium glutamicum, examine purified PQO, and describe the crystal structure of the native enzyme and a truncated version. The specific PQO activity was highest in stationary phase cells grown in complex medium, lower in cells grown in complex medium containing glucose or acetate, and lowest in cells grown in minimal acetate-medium. A similar pattern with about 30-fold higher specific PQO activities was observed in C. glutamicum with plasmid-bound pqo expression under the control of the tac promoter, indicating that the differences in PQO activity are likely due to post-transcriptional control. Continuous cultivation of C. glutamicum at dilution rates between 0.05 and 0.4 h-1 revealed a negative correlation between PQO activity and growth rate. Kinetic analysis of PQO enzymes purified from cells grown in complex or in minimal acetate-medium revealed substantial differences in specific activity (72.3 vs. 11.9 U·mg protein-1) and turnover number (kcat: 440 vs. 78 s-1, respectively), suggesting post-translational modifications affecting PQO activity. Structural analysis of PQO revealed a homotetrameric arrangement very similar to the Escherichia coli pyruvate oxidase PoxB except for the C-terminal membrane binding domain, which exhibited a conformation markedly different from its PoxB counterpart. A truncated PQO variant lacking 17 C-terminal amino acids showed higher affinity to pyruvate and was independent of detergent activation, highlighting the importance of the C-terminus for enzyme activation and lipid binding.