Urease Enzyme Activity Research Articles

Antioxidant and anti-urease activities of Cardamine bulbifera: Insights from molecular docking and density functional theory studies

IntroductionCardamine bulbifera (L.) Crantz, commonly known as “Asian bittercress”, is a plant species widely distributed across Asia. It belongs to the Brassicaceae and has been traditionally used in medicinal applications. This study investigates the phytochemical composition, phenolic content, antioxidant properties, and anti-urease activity of C. bulbifera. MethodsIn this study, the aerial parts of the C. bulbifera were collected. The total polyphenol content of the extracts was determined using spectrophotometric methods, and its antioxidant and anti-urease activities were assessed. The chemical composition was characterised using gas chromatography-mass spectroscopic (GC-MS) analysis. Additionally, molecular docking studies explored potential interactions between the identified compounds and Helicobacter pylori CagA oncoprotein. Furthermore, Density Functional Theory (DFT) analysis provided valuable insights into the electronic properties of the main compounds. ResultThe total phenolic content of C. bulbifera extract was 225 ± 0.02 mg GAE/g extract DW, and the total flavonoid content was 62.64 ± 3.27 mg QE/g extract DW. The 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) assay revealed an IC50 value of 1.01 ± 0.04 mg/mL, indicating significant antioxidant activity. The IC50 value for anti-urease activity was determined to be 2.74 ± 0.03 µg/mL, suggesting potent inhibition of urease enzyme activity. GC-MS analysis identified 15 bioactive compounds in the extract. Molecular docking studies highlighted Phenol, 3,5-bis(1,1-dimethylethyl) and 1-dodecanol as compounds with the highest binding affinity for the H. pylori CagA oncoprotein, suggesting potential therapeutic implications against H. pylori-related cancers. Additionally, DFT analysis emphasised these compounds' electronic properties and chemical reactivities, indicating their potential role in future pharmaceutical developments. ConclusionC. bulbifera exhibits rich phenolic and flavonoid content and significant antioxidant and intense urease inhibition activities. These findings suggest that C. bulbifera may offer potential therapeutic options for conditions related to H. pylori infections, including cancer. Further research is needed to explore its mechanisms of action and clinical applications in more detail.

Productivity and health of soil cultivated with forage cactus (Opuntia cochenillifera (L.) Mill.) with combined use of topsoil and biochar

Agricultural production in semi-arid environments represents a significant challenge, as these are areas that have soils with limited nutrient availability and scarcity of precipitation, which require adaptive and innovative strategies. The application of biochar to improve soil quality and increase productivity has been well studied. However, there is still a gap regarding the use of biochar with the technique of incorporating different topsoils on plant development and soil health. This technique has emerged as an alternative for producers to increase soil fertility. This technique is emerging as an alternative for producers to enhance soil fertility. The hypothesis of the present study is that the use of biochar associated with the topsoil technique increases the productivity and yield of cactus, mainly by changing the properties of the soil and its microbiological interactions. To test this hypothesis, we used different sources of biochar (bean husk, coffee grounds and soursop residue) in association with two types of topsoil (Acrisol and Luvisol). Our analyzes revealed changes on plants development, chemical and microbiological soil attributes. The topsoil technique significantly interfered with mean cladode length (COM), stem diameter (DI), plant height (AL), pH, Na+, K+ and urease enzymatic activity, while biochar had a significant effect on microbial respiration rate. In general, our results expand the knowledge about the use of biochar associated with topsoil, suggesting it as a promising, low-cost way that guarantees the conservation and health of the agroecosystem, contributing with important information for the development of alternative technologies to conventional ones.

Role of compost in assisted phytostabilization via three naturally occurring species on mine-contaminated soil and health risk alleviation for livestock

Phytostabilization emerges as an efficient and enduring remediation technique for heavy metal-contaminated mine soils using plants acclimatized to such circumstances. In this research, the synergistic effects of Marrubium cuneatum, Verbascum speciosum, and Stipa arabica species, alongside municipal solid waste compost (MSWC) amendment at 0, 1, 3, and 5 % rates, were assessed for a six-month pot experiment aimed at remediating naturally polluted soil containing lead (Pb) and cadmium (Cd) and mitigating the health risks associated with these metal exposures for grazing ruminants. Applying compost improved the aboveground biomass of M. cuneatum and V. speciosum and S. arabica by 13, 19, and 18 %, respectively. Also, soil dehydrogenase and urease enzyme activities were enhanced up to 131 and 34 %. Upon MSWC application, all three species uptake Cd and Pb at a significantly lower rate, except in the V. speciosum shoot at 3 and 5 % doses, and bioaccumulation factors were markedly diminished. Compost notably augmented the antioxidant system, leading to a reduction in malondialdehyde content of V. speciosum, M. cuneatum, and S. arabica by 35, 29, and 23 %, respectively, and an elevation in chlorophyll level. Superoxide dismutase, catalase, glutathione peroxidase, and ascorbate peroxidase activities either increased or displayed no significant differences. Risk assessment revealed that MSWC substantially decreased the daily intake of Pb and Cd and their accumulation in animal organs, thereby eliminating limits on meat consumption. Findings affirm the combined efficacy of MSWC and the study species in stabilizing contaminated Pb and Cd soils, diminishing risks to animal feed security and human health.

Assessment of Soil Enzyme Activities in Plant Root Zone of Saline Soil Reclaimed by Drip Irrigation with Saline Groundwater

Drip irrigation with saline water is frequently adopted to realize the sustainable utilization of saline–sodic soil with high water tables, and soil enzyme activities can be used to indicate changes in soil quality. In the current study, spatiotemporal changes in soil urease enzyme (URE), alkaline phosphatase (ALP) and invertase (INV) activities were investigated during consecutive growing seasons. Soil in beds was sampled before planting (0 y) and one, two, three and four years after the growing season (1 y, 2 y, 3 y, 4 y), and these samples were distributed at four horizontal distances from the drip line (0, 10, 20 and 30 cm) and four vertical soil depths (0–10, 10–20, 20–30 and 30–40 cm). The results showed that a distribution pattern of URE and ALP activities formed during the first growing season, while the distribution of INV activity formed until the third growing season. All three soil enzyme activities in the upper soil layers and positions close to the drip line were more greatly affected by planting year. The average URE activity of the soil profile decreased slightly during the first year and increased by about 220% and decreased by 20% after reclamation for two and three years, and finally, it increased to 4.9 μg NH4+·g−1·h−1 at the end of the fourth growing season. ALP activity remained stable during the first two years and rapidly increased in the following years; in particular, in the fourth year, it reached 32.7 μg ph(OH)·g−1·h−1. INV activity increased continually with the number of years after planting and reached 1009.0 μg glu·g−1·h−1 at the fourth season’s end. An analysis of variance indicated that URE, ALP and INV activities varied insignificantly among the time points of 0 y, 1 y, 2 y and 3 y (p < 0.05), while they were significantly higher for 4 y than for 0 y and 1 y. In addition, all three enzyme activities of the soil profile had an exponentially increasing trend with the number of years after planting. These results indicated the soil quality in saline–sodic soils could be improved with time under drip irrigation with local saline groundwater, especially around the drip line.

The Effects of the Co-Application of MCPA Herbicide and Urea on Grass Rhizosphere Microcosms

Background: Urea fertilizer and MCPA herbicide are widely used agrochemicals in pastures. Even though urea hydrolysis impacts soil pH, potentially affecting MCPA dissipation, little is known about the effects of their co-application into the rhizosphere. Hence, we aimed to analyze the dynamics of urea transformation and MCPA dissipation when both are co-applied to the soil. Methods: A greenhouse experiment was conducted with a planted control and treatments incorporating urea and/or MCPA. Subsequently, pH changes, urea transformation into N-NH4+ and N-NO3−, the enzymatic activity of urease and dehydrogenase, and MCPA dissipation were monitored for 30 d. Results: Urea application induced a significant (p < 0.05) pH change, production of N-NH4+ (from 50 and 250 mg kg−1) and N-NO3− (from 206 to 347 mg kg−1), and urease (from 12 to 35 µmol N-NH4+g−1 h−1) and dehydrogenase (from 0.5 to 2.5 mg TPF g−1 h−1) activities. Urea also decelerated MCPA dissipation in the latter half of the experiment, whereas MCPA reduced urease activity when urea and herbicide were co-applied. Conclusions: Urea was the primary factor modifying the properties of the rhizosphere by stimulating the activity of microbial enzymes, shaping the pH changes during its mineralization, and decelerating MCPA dissipation. MCPA did not reduce urea mineralization but slowed urease activity, constituting an insight that requires further study.

Nano silica’s role in regulating heavy metal uptake in Calendula officinalis

BackgroundSoil contamination with heavy metals poses a significant threat to plant health and human well-being. This study explores the potential of nano silica as a solution for mitigating heavy metal uptake in Calendula officinalis.ResultsGreenhouse experiments demonstrated, 1000 mg•kg− 1 nano silica caused a 6% increase in soil pH compared to the control treatment. Also in 1000 mg. kg− 1 nano silica, the concentrations of available Pb (lead), Zn (zinc), Cu (copper), Ni (nickel), and Cr (chromium) in soil decreased by 12%, 11%, 11.6%, 10%, and 9.5%, respectively, compared to the control. Nano silica application significantly reduces heavy metal accumulation in C. officinalis exposed to contaminated soil except Zn. In 1000 mg.kg− 1 nano silica shoots Zn 13.28% increased and roots Zn increased 13% compared to the control treatment. Applying nano silica leads to increase the amount of phosphorus (P) 25%, potassium (K) 26% uptake by plant, In 1000 mg.kg − 1 treatment the highest amount of urease enzyme activity was 2.5%, dehydrogenase enzyme activity, 23.6% and the highest level of alkaline phosphatase enzyme activity was 13.5% higher than the control treatment.ConclusionNano silica, particularly at a concentration of 1000 mg.kg − 1, enhanced roots and shoots length, dry weight, and soil enzyme activity Moreover, it increased P and K concentrations in plant tissues while decreasing heavy metals uptake by plant.

Short-Term Response of Soil Microbial Community and Soil Biochemical Properties to Soybean Intercropping in a Cassava-Based Cropping System

A cassava-based cropping system study assessed the short-term response of soil chemical and biochemical properties and soil microbial communities to soybean intercropping. Two soybean varieties (‘Select Manchuria’ and ‘Tiwala 12’) were tested as intercrops. The treatments did not affect the soil’s chemical (pH, OM, N, P, K) and biochemical (basal respiration and dehydrogenase and urease enzyme activities) properties after one cropping period. Amplicon sequencing analysis found that intercropping promoted the abundance of bacterial orders Actinomycetales, Solibacterales, Sphingomonadales, and Rhodospirillales. These groups play active roles in organic matter decomposition and can potentially improve the soil quality in an area by enhancing the soil’s organic matter content. In terms of community analysis, principal coordinate analysis (PCoA) and redundancy analysis (RDA) showed separation between mono and intercropping systems, while ANOSIM and PEMANOVA did not detect any significance between varieties and cropping systems for both bacterial and fungal data. These results suggest that, in the short term, introduction of soybean in a cassava-based cropping system affects selected microbial groups, but the overall influence in the microbial community is not distinctly detected.

Phellodendron chinense C.K.Schneid: An in vitro study on its anti-Helicobacter pylori effect

Ethnopharmacological relevancePhellodendron chinense C.K.Schneid(P. chinense Schneid) is known in TCM as Huang Bo, is traditionally used to support gastrointestinal function and alleviate stomach-related ailments, including gastric ulcer bleeding and symptoms of gastroesophageal reflux disease. Helicobacter pylori (H. pylori) is classified by the WHO as a Group 1 carcinogen. However, the specific activity and mechanism of action of P. chinense Schneid against H. pylori infection remain unclear. It has been noted that Huangjiu processing may alter the bitter and cold properties of P. chinense Schneid, but its effect on antimicrobial activity requires further investigation. Additionally, it remains uncertain whether berberine is the sole antimicrobial active component of P. chinense Schneid. Aim of studyThis study aims to elucidate the anti-H. pylori infection activity of P. chinense Schneid, along with its mechanism of action and key antimicrobial active components. Materials and methodsPhytochemical analysis was carried out by UPLC-MS/MS. HPLC was employed to quantify the berberine content of the extracts. Antimicrobial activity was assessed using the micro broth dilution method. Morphology was observed using SEM. The impact on urease activity was analyzed through in vitro urease enzyme kinetics. RT-qPCR was employed to detect the expression of virulence genes, including adhesin, flagellum, urease, and cytotoxin-related genes. The adhesion effect was evaluated by immunofluorescence staining and agar culture. ResultsP. chinense Schneid exhibited strong antimicrobial activity against both antibiotic-sensitive and resistant H. pylori strains, with MIC ranging from 40 to 160 μg/mL. Combination with amoxicillin, metronidazole, levofloxacin, and clarithromycin did not result in antagonistic effects. P. chinense Schneid induced alterations in bacterial morphology and structure, downregulated the expression of various virulence genes, and inhibited urease enzyme activity. In co-infection systems, P. chinense Schneid significantly attenuated H. pylori adhesion and urease relative content, thereby mitigating cellular damage caused by infection. Huangjiu processing enhanced the anti-H. pylori activity of P. chinense Schneid. Besides berberine, P. chinense Schneid contained seven other components with anti-H. pylori activity, with palmatine exhibiting the strongest activity, followed by jatrorrhizine. ConclusionsThis study sheds light on the potential therapeutic mechanisms of P. chinense Schneid against H. pylori infection, demonstrating its capacity to disrupt bacterial structure, inhibit urease activity, suppress virulence gene transcription, inhibit adhesion, and protect host cells. The anti-H. pylori activity of P. chinense Schneid was potentiated by Huangjiu processing, and additional components beyond berberine were identified as possessing strong anti-H. pylori activity. Notably, jatrorrhizine, a core component of P. chinense Schneid, exhibited significant anti-H. pylori activity, marking a groundbreaking discovery.

Arbuscular mycorrhiza and rhizosphere soil enzymatic activities as modulated by grazing intensity and plant species identity in a semi-arid grassland

Understanding the symbiosis of arbuscular mycorrhizal fungi (AMF) with plants in relation to soil nutrients and enzyme activities under different grazing intensities can be an important guide for the management and protection of semi-arid grasslands. The aim of the present study was to evaluate how the interaction of grazing intensity and plants shapes the composition of AMF communities and enzyme activities in a semi-arid grassland ecosystem in Iran. Sampling focused three dominant plant species (i.e., Salsola laricina, Artemisia siberia, and Stipa hohenackeriana) at sites with different grazing intensities. Soil chemical properties, enzyme activities, root colonization by AMF and AMF communities in the roots were evaluated. Potassium and nitrogen, as well as alkaline phosphatase and urease enzymatic activities were significantly increased at the heavily grazed site, whereas root colonization by AMF was reduced by the high grazing intensity. In addition, AM fungal root colonization is dependent on the host plant species and easier to measure as a sensitive indicator of sustainable grazing. Neither plant species nor grazing intensity affected AM fungal diversity in roots, which could be due to the overall low phylogenetic diversity of AMF in the grassland and the lack of significant differences in soil humidity, pH and organic carbon between the sites. However, plant species and soil properties were the two factors explaining variation in AMF community composition, while grazing had no significant effect. Therefore, AMF communities in root of the semi-arid grassland plants responded largely to plant type rather than to grazing intensity.

The impact on Cd bioavailability and accumulation in rice (Oryza sativa L.) induced by dry direct-seeding cultivation method in field-scale experiments

Dry direct-seeded rice cultivation has gained popularity and expanded its cultivated area due to reduced labor requirements and water consumption. However, the impact of this cultivation method on cadmium (Cd) bioavailability in soil and the accumulation levels in grains remains uncertain. Field experiments were conducted in acidic soils at two locations in southern China to compare rice varieties and evaluate the dry direct-seeding method alongside the wet direct-seeding and traditional transplanting methods. Dry direct-seeded rice reached significantly higher Cd concentrations in its tissues starting from the heading stage than transplanted rice. Cd accumulation levels by the maturation stage in the brown rice of dry direct-seeded rice were 18.33 %–150.69 % higher than those of wet direct-seeded and transplanted rice, with a considerable ability to translocate Cd into brown rice. Furthermore, dry direct seeding decreased iron plaque formation, particularly in the amorphous Fe form; it resulted in high soil temperature and low moisture content during tillering, elevating Cd availability in the soil. Additionally, the proportion of ions and more labile forms of Cd in the soil solution was high. Moreover, the soil under dry direct seeding had high urease and acid phosphatase enzyme activities. However, low richness and diversity in the bacterial community were characterized by a significant increase in the relative abundance of Actinobacteria and Gammaproteobacteria at the class level, while exhibiting decreased relative abundances of Alphaproteobacteria, Bacilli, and KD4-96, along with fewer biomarkers. Nonetheless, these differences are gradually reduced during the maturation stage. Overall, although dry direct seeding offers several advantages, it is crucial to implement additional measures to mitigate the increased health risks linked to rice cultivation through this approach in Cd-contaminated areas.

Removal of cadmium through biomineralization using halophilic and ureolytic bacteria under saline conditions

Heavy metal pollution poses an immense challenge to humanity owing to its severe toxic effects on living beings. This study investigates the bioremediation potential of the urease-hydrolyzing and halophilic bacterium Bacillus paramycoides- MSR1 to reduce the toxicity of cadmium (Cd) in the environment under salt stress through microbially induced calcium carbonate precipitation (MICCP). The enzymatic activity of urease and calcium carbonate (CaCO3) precipitation was studied under different concentrations of salt stress (0%, 3.5%, 5%, 7.5%, 10%, 15%) and maximum urease activity of 735.7 U/ml was recorded at 5% salt stress. The urease activity and CaCO3 precipitation decreased with increasing Cd toxicity (0, 10, 20, 30, 40, 50, and 60 μM). The maximum concentration of Cd endured by bacteria was determined by IC50 value and the minimum inhibitory concentration of Cd was recorded as 9.86 μM under 5% stress. Atomic absorption spectroscopy results revealed that Cd removal was as high as 90.3% under 5% salt stress. Microstructural analysis through FE-SEM revealed the surface topography of carbonate crystals as rhombohedral, whereas EDS confirmed the presence of CaCO3 and Cd in the bio-precipitates. These results suggest that MICCP is a potential, environmentally safe, low-cost technique and an excellent alternative to conventional heavy metal removal strategies from the environment.

Influence of enzyme activity on domestic wastewater amelioration in a hybrid subsurface flow constructed wetland.

Wastewater treatment in a constructed wetland is achieved by the presence of plant species, the metabolism of microorganisms, and the enzyme activities. Three small-scale hybrid subsurface flow constructed wetlands (HSFCWs) planted with Arundo donax and one unplanted HSFCW were constructed near a water resource recovery facility at Guru Gobind Singh Indraprastha University. The purpose of the study was to determine the correlation between soil enzymatic activities and the removal of contaminants from domestic wastewater. Enzyme activity of phosphatase, protease, urease, and cellulase increased with an increase in temperature. A strong correlation between enzyme activities and TKN and surfactant removal was observed, whereas moderate correlation was observed with phosphate in planted HSFCW during the study. The correlation between COD removal and enzyme activities was low to moderate. In unplanted HSFCW, the correlation between enzyme activities and COD removal was negative, negligible to moderate to strong in the case of TKN, low to moderate in the case of phosphate, and negligible to low in the case of surfactants. The increased removal efficiency of the planted system compared with that of the unplanted system indicated a positive impact on enzyme activities with the growth of plants and their roots. PRACTITIONER POINTS: Protease, urease, and cellulase activities: Planted HSFCW exhibited higher protease, urease, and cellulase activities than unplanted, signifying enhanced breakdown. July displayed maximum enzyme activities, correlating with heightened biological breakdown in both systems. Fluctuations in enzyme activities reflected seasonal changes, influencing nutrient degradation rates. Planted HSFCW consistently showed higher enzymatic activities across protease, urease, and cellulase than unplanted.

Biogenic calcium improved Cd2+ and Pb2+ immobilization in soil using the ureolytic bacteria Bacillus pasteurii

Bioremediation based on microbial-induced carbonate precipitation (MICP) was conducted in cadmium and lead contaminated soil to investigate the effects of MICP on Cd and Pb in soil. In this study, soil indigenous nitrogen was shown to induce MICP to stabilize heavy metals without inputting exogenous urea. The results showed that applying Bacillus pasteurii coupled with CaCl2 reduced Cd and Pb bioavailability, which could be clarified through the proportion of exchangeable Cd and Pb in soil decreasing by 23.65 % and 12.76 %, respectively. Moreover, B. pasteurii was combined separately with hydroxyapatite (HAP), eggshells (ES), and oyster shells (OS) to investigate their effects on soil heavy metals' chemical fractions, toxicity characteristic leaching procedure (TCLP)-extractable Cd and Pb as well as enzymatic activity. Results showed that applying B. pasteurii in soil significantly decreased the heavy metals in the exchangeable fraction and increased them in the carbonate phase fraction. When B. pasteurii was combined with ES and OS, the content of carbonate-bound Cd increased by 114.72 % and 118.81 %, respectively, significantly higher than when B. pasteurii was combined with HAP, wherein the fraction of carbonate-bound Cd increased by 86 %. The combination of B. pasteurii and biogenic calcium effectively reduced the leached contents of Cd and Pb in soil, and the TCLP-extractable Cd and Pb fractions decreased by 43.88 % and 30.66 %, respectively, in the BP + ES group and by 52.60 % and 41.77 %, respectively, in the BP + OS group. This proved that MICP reduced heavy metal bioavailability in the soil. Meanwhile, applying B. pasteurii and calcium materials significantly increased the soil urease enzyme activity. The microstructure and chemical composition of the soil samples were studied, and the results from scanning electron microscope, Fourier transform infra-red spectroscopy, and X-ray diffraction demonstrated the MICP process and identified the formation of CaCO3, Ca0.67Cd0.33CO3, and PbCO3 in heavy metal-contaminated soil.

Multiomics Explore the Detoxification Mechanism of Nanoselenium and Melatonin on Bensulfuron Methyl in Wheat Plants.

Combining nanoselenium (nano-Se) and melatonin (MT) was more effective than treatment alone against abiotic stress. However, their combined application mitigated the toxic effects of bensulfuron methyl, and enhanced wheat growth and metabolism has not been studied. Metabolomics and proteomics revealed that combining nano-Se and MT markedly activated phenylpropanoid biosynthesis pathways, elevating the flavonoid (quercetin by 33.5 and 39.8%) and phenolic acid (vanillic acid by 38.8 and 48.7%) levels in leaves and roots of wheat plants. Interstingly, beneficial rhizosphere bacteria in their combination increased (Oxalobacteraceae, Nocardioidaceae, and Xanthomonadaceae), which positively correlated with the enhancement of soil urease and fluorescein diacetate enzyme activity (27.0 and 26.9%) and the allelopathic substance levels. To summarize, nano-Se and MT mitigate the adverse effects of bensulfuron methyl by facilitating interactions between the phenylpropane metabolism of the plant and the beneficial microbial community. The findings provide a theoretical basis for using nano-Se and MT to remediate herbicide-contaminated soil.

Atividade biológica do solo cultivado com feijão-guandu sob diferentes manejos de adubação

ABSTRACT: Fertilization management of pigeon peas can increase soil quality and the N utilization by plants. Therefore, we evaluated the biological activity of soil cultivated with pigeon peas (Cajanus cajan (L) Millsp.) under different fertilization treatments. A randomized block design was used with three replicates and a 3×5 factorial arrangement (genotype×fertilization and inoculation management). At full flowering stage, the plants were collected and shoot dry matter was evaluated. Soil was sampled at 0-20 cm for various analysis viz. nodulation assessment; soil organic carbon; total nitrogen; carbon and nitrogen from microbial biomass; C/N ratio; β-glucosidase and urease enzymatic activity. Data were subjected to analysis of variance and Tukey’s test (P ≤ 0.05). A Pearson correlation matrix was constructed, and the similarity between treatments was evaluated using the Mahalanobis distance and grouping was done using the unweighted pair group method with arithmetic average (UPGMA). Both the experimental genotypes (BRS03 and BRS04) showed similar nodulation and shoot dry matter pattern with respect to fertilizer treatments. Furthermore, the microbial inoculation promoted a higher shoot dry matter content in all the genotypes. The application of mineral N and inoculation increased the total N content in the soil, favoring the mineralization of this nutrient. During the testing phase, the genotypes exhibited an increase in microbial carbon and microbial quotient levels, indicating an improvement in soil quality. The combination of fertilization and inoculation increased the enzymatic activity of β-glucosidase and urease. The correlation matrix showed a strong association between N total and C/N ratio. The formation of groups by UPGMA was observed as a function of inoculation, demonstrating its effect on soil biological variables.

N-Adamantanyl-2-(2-(phenyl)hydrazone)-3-oxobutanamides endowed with dual inhibitors of urease and α-glucosidase: Design, synthesis, and computational studies

The conventional approach of drug development, which focused on inhibiting a single target, has been superseded by a more advanced strategy known as multi-target design. In this study, we describe the synthesis of new N-adamantyl-2-(2-(phenyl)hydrazone)-3-oxobutanamide (6a-k). These compounds were designed with the intention of serving as prospective drug like candidates that can target both alpha glucosidase and urease enzymes. For this purpose, both in-vitro and in-silico investigations were performed after synthesis and characterization. The density functional theory calculations were employed to calculate optimized geometries, global reactivity descriptors and frontier molecular orbital (FMO) analysis. All compounds were found reactive and compounds 6b, 6c, 6d and 6e were found the most stable. The synthesized compounds were also tested for their ability to inhibit the enzyme activities of urease and α-glucosidase. The compound 6c exhibited strong inhibition of urease enzyme, with IC50 value of 13.10 ± 0.55 µM, in comparison to the IC50 value of the standard inhibitor thiourea i.e., 16.4 ± 1.02 µM. In addition to this, compounds 6d and 6e demonstrated a significant α-glucosidase inhibition with IC50 values of 17.16 ± 0.91 µM but found less potent as compared to the standard inhibitor Acarbose, i.e. 9.80 ± 0.20 μM. The structure–activity relationship (SAR) was established and the in-vitro results were further supported by the molecular docking investigations and molecular dynamic simulation studies. The in-vitro and in-silico results demonstrated a strong correlation in assessing the drug-like characteristics of all synthesized compounds (6a-k). The in silico investigation confirmed the findings in the search for inhibitors against the listed enzymes by elucidating the binding relationship between most of the active compounds and the active site of urease and α-glucosidase.

Reduction of UreB and CagA expression level by siRNA construct in Helicobacter pylori strain SS1

BackgroundTwo important virulence factors, urease and cagA, play an important role in Helicobacter pylori (H. pylori) gastric cancer. Aim of this study was to investigate the expression level and function of ureB and cagA using small interfering RNAs (siRNA).MethodsSS1 strain of H. pylori was considered as host for natural transformation. siRNA designed for ureB and cagA genes were inserted in pGPU6/GFP/Neo siRNA plasmid vector to evaluate using phenotypic and genotypic approaches. Then, qPCR was performed for determining inhibition rate of ureB and cagA gene expression.ResultsThe expression levels of siRNA-ureB and siRNA-cagA in the recombinant strain SS1 were reduced by about 5000 and 1000 fold, respectively, compared to the native H. pylori strain SS1. Also, preliminary evaluation of siRNA-ureB in vitro showed inhibition of urea enzyme activity. These data suggest that siRNA may be a powerful new tool for gene silencing in vitro, and for the development of RNAi-based anti-H. pylori therapies.ConclusionOur results show that targeting ureB and cagA genes with siRNA seems to be a new strategy to inhibit urease enzyme activity, reduce inflammation and colonization rate.

Effects of natural and prolonged crude oil pollution on soil enzyme activities

In this study the effects of natural and long-term oil pollution levels (H: high, M: moderate and L: low) on the activity of lipase, β-glucosidase, urease, phosphatase, and dehydrogenase were analyzed. For this purpose, 120 oil-contaminated soil samples were collected from four locations located in oil-rich region of the west of Iran (Kermanshah province, Iran). After determining the physicochemical characteristics of soils, microbial counting and enzyme activities were measured. To determine the total bacterial population and bacteria involved in crude oil decomposition, bacterial counting was done in NA and CFMM culture media, respectively, which had a direct relationship with the increase in crude oil content. The average percentage of crude oil measured by Soxhlet method was 4.03%, 9.95% and 22.50% respectively for L, M and H levels. The results showed that the bacterial population increased with the increase of the contamination intensity. All the studied enzymes, except urease, had the highest activity in heavily polluted soils (H soil), and their lowest activity was observed in lightly polluted soils (L soils). Location 1 had the highest bacterial population as well as the highest activity of dehydrogenase (52.13 μg TPFg−1 h−1), acid phosphatase (35.86 μg PNPg−1 h−1), alkaline phosphatase (78.13 μg PNPg−1 h−1), lipase (92.58 μ MLA g−1 h−1) and β-glucosidase (21.67 μg PNPg−1 h−1) enzymes. In the soil samples of location 4, which had the lowest number of bacteria, the highest activity of urease enzyme (592.36 μg NH4g−1 h−1) was obtained. Principal components analysis (PCA) was also performed and 67% of the cumulative of the samples could be explained by the first two components (biochemical and physical). The findings of this research showed that natural and prolonged crude oil pollution caused the selection of oil-resistant microbial communities, and therefore we saw their positive response to the presence of oil compounds and increased enzyme activity other than urease activity.

Effect of proteins on biocementation in construction materials

This study examines the effect of proteins on the binding property and microstructure of enzymatic-induced calcium carbonate precipitation (EICP) in cementitious environment. The protein modified precipitates generally demonstrated improved binding to a glass slide surface or cement paste surface compared to the control precipitate. A marked decrease in the amount and binding strength of the precipitates in the cementitious environment was observed due to a reduction in the urease enzyme activity. The protein modified precipitates exhibited noticeable improvement compared to the control precipitate in cementitious environment which could arise from the ability of the proteins to partially shield urease from the negative effect of high pH. The protein gel network formation due to the complexation between the proteins and Ca2+ provides nucleation sites for CaCO3 crystallization. The FTIR, SEM, TGA, and XRD results indicated that vaterite is the dominant polymorph in cementitious environment compared to calcite in deionized water.

Manufacture and Characterization of PVA-Enzyme/GA/PANI-HCl or PANI-p-toluentsulfonate/PVC-KTpClPB-o-NPOE Indicator Electrode Membranes, Analysis, XRD, SEM-EDX and FTIR

This research aims at the synthesis and characterization of indicator electrode membranes consisting of PVA-Enzyme/GA/PANI/PVC-KTpClPB-o-NPOE, urea analyte. PANI-HCl and PANI-p-toluensulfonate conducting polymers, 61% and 66% o-NPOE plasticizer variations in PVC-KTpClPB, as well as urease enzyme activity in one or three drops of 0.5 mL PVA (50% water: 50% ethanol). The biosensor potentiometric method was used with the technique of immobilizing the urea enzyme which is the urea analyte. The multi-membrane indicator electrode is PVA-Enzyme/GA/PANI/PVC-KTpClPB-o-NPOE, PANI dissolved in HCl and p-toluensulfonic acid respectively at a concentration of 6 M and 2 M. Each membrane on the indicator electrode is coated with one time. Results of XRD analysis of the spectrum pattern of the PANI-HCl indicator electrode with intensities of 4400 a.u, 1386 a.u, 1724 a.u with a 2theta angle of 28.6 degrees, PANI-p-toluensulfonate with intensities of 10940 a.u, 9194 a.u, 5312 a.u with a 2theta angle of 18.2 degrees. SEM-EDX analysis showed differences in the morphology of the o-NPOE plasticizer of 61% and 66% as well as an increase in cps/eV from the number of drops of the urease enzyme, one drop was lower than three drops. FTIR analysis shows an increase in transmittance by PANI towards PPy. Analysis of the properties of a multi-membrane indicator electrode with one layer of the best sample is PANI-p-toluensulfonate