Fruit and vegetable waste used as bacterial growth media for the biocementation of two geomaterials.

This paper investigates the feasibility of using randomly collected fruit and vegetable (FV) waste as a cheap growing medium of bacteria for biocementation applications. Biocementation has been proposed in the literature as an environmentally-friendly ground improvement method to increase the stability of geomaterials, prevent erosion and encapsulate waste, but currently suffers from the high costs involved, such as bacteria cultivation costs. After analysis of FV waste of varied composition in terms of sugar and protein content, diluted FV waste was used to grow ureolytic (S. pasteurii, and B.licheniformis) and also an autochthonous heterotrophic carbonic anhydase (CA)-producing B.licheniformis strain, whose growth in FV media had not been attempted before. Bacterial growth and enzymatic activity in FV were of appropriate levels, although reduced compared to commercial media. Namely, the CA-producing B.licheniformis had a maximum OD600 of 1.799 and a CA activity of 0.817 U/mL in FV media. For the ureolytic pathway, B. licheniformis reached a maximum OD600 of 0.986 and a maximum urease activity of 0.675 mM urea/min, and S. pasteurii a maximum OD600 = 0.999 and a maximum urease activity of 0.756 mM urea/min. Biocementation of a clay and locomotive ash, a geomaterial specific to UK railway embankments, using precultured bacteria in FV was then proven, based on recorded unconfined compressive strengths of 1-3 MPa and calcite content increases of up to 4.02 and 8.62 % for the clay and ash respectively. Scanning Electron Microscope (SEM) and energy dispersive X-ray spectroscopy (EDS), attested the formation of bioprecipitates with characteristic morphologies and elementary composition of calcite crystals. These findings suggest the potential of employing FV to biocement these problematic geomaterials and are of wider relevance for environmental and geoenvironmental applications involving bioaugmentation. Such applications that require substrates in very large quantities can help tackle the management of the very voluminous fruit and vegetable waste produced worldwide.

Inhibitory effect of polyphenols from sumac, pomegranate and indian almond on urease producing bacteria and jack bean urease activity.

Urinary tract infection caused by Klebsiella, Proteus and Streptococcus is a urease dependent process, hence treatment of these infections by antibacterial compounds lies in inhibition of their virulence factors. The crude methanolic extracts derived from sumac fruit, pomegranate peel and Indian almond leaves were separated into anthocyanin and non-anthocyanin fractions using solid phase cartridges. The inhibitory effect of these fractions was determined on the growth of urease producing species and jack bean urease activity. Known compounds in the fractions were also docked with ureases of different biological origins viz. K. pneumoniae (PDB ID: 8HCN), K. aerogenes (PDB ID: 2KAU), Helicobacter pylori (PDB ID:8HC1)and Canavalia ensiformis (jack bean) (PDB ID: 3LA4) to determine their binding affinities and interaction with the enzyme. All the fractions showed significant inhibition growth for P. mirabilis, S. epidermidis and K.pneumoniae. Among the samples, sumac showed greatest inhibition against all (MIC 6-25 mg.mL-1) while among the fractions, anthocyanin was found to be most active (MIC 6-12 mg/mL). Likewise, all fractions inhibited urease with lowest ICs50 shown by sumac fractions (21-116 µg.mL-1). Out of 39 compounds docked, 27 showed interaction with movable flaps and/or active site of ureases which explains their mode of inhibition.

Effects of enzyme-induced carbonate precipitation technique on multiple heavy metals immobilization and unconfined compressive strength improvement of contaminated sand.

Enzyme-induced carbonate precipitation (EICP) has been studied in remediation of heavy metal contaminated water or soil in recent years. This paper aims to investigate the immobilization mechanism of Zn2+, Ni2+, and Cr(VI) in contaminated sand, as well as strength enhancement of sand specimens by using EICP method with crude sword bean urease extracts. A series of liquid batch tests and artificially contaminated sand remediation experiments were conducted to explore the heavy metal immobilization efficacy and mechanisms. Results showed that the urea hydrolysis completion efficiency decreased as the Ca2+ concentration increased and the heavy metal immobilization percentage increased with the concentration of Ca2+ and treatment cycles in contaminated sand. After four treatment cycles with 0.5 mol/L Ca2+ added, the immobilization percentage of Zn2+, Ni2+, and Cr(VI) were 99.99 %, 86.38 %, and 75.18 %, respectively. The microscale analysis results presented that carbonate precipitates and metallic oxide such as CaCO3, ZnCO3, NiCO3, Zn(OH)2, and CrO(OH) were generated in liquid batch tests and sand remediation experiments. The SEM-EDS and FTIR results also showed that organic molecules and CaCO3 may adsorb or complex heavy metal ions. Thus, the immobilization mechanism of EICP method with crude sword bean urease can be considered as biomineralization, as well as adsorption and complexation by organic matter and calcium carbonate. The unconfined compressive strength of EICP-treated contaminated sand specimens demonstrated a positive correlation with the increased generation of carbonate precipitates, being up to 306 kPa after four treatment cycles with shear failure mode. Crude sword bean urease with 0.5 mol/L Ca2+ added is recommended to immobilize multiple heavy metal ions and enhance soil strength.

Microbially induced carbonate precipitation with Arthrobacter creatinolyticus: An eco-friendly strategy for mitigation of chromium contamination.

Chromium contamination from abandoned industrial sites and inadequately managed waste disposal areas poses substantial environmental threat. Microbially induced carbonate precipitation (MICP) has shown promising, eco-friendly solution to remediate Cr(VI) and divalent heavy metals. In this study, MICP was carried out for chromium immobilization by an ureolytic bacterium Arthrobacter creatinolyticus which is capable of reducing Cr(VI) to less toxic Cr(III) via extracellular polymeric substances (EPS) production. The efficacy of EPS driven reduction was confirmed by cellular fraction analysis. MICP carried out in aqueous solution with 100 ppm of Cr(VI) co-precipitated 82.21% of chromium with CaCO3 and the co-precipitation is positively correlated with reduction of Cr(VI). The organism was utilized to remediate chromium spiked sand and found that MICP treatment decreased the exchangeable fraction of chromium to 0.54 ±â€¯0.11% and increased the carbonate bound fraction to 26.1 ±â€¯1.15% compared to control. XRD and SEM analysis revealed that Cr(III) produced during reduction, influenced the polymorph selection of vaterite during precipitation. Evaluation of MICP to remediate Cr polluted soil sample collected from Ranipet, Tamil Nadu also showed effective immobilization of chromium. Thus, A. creatinolyticus proves to be viable option for encapsulating chromium contaminated soil via MICP process, and effectively mitigating the infiltration of Cr(VI) into groundwater and adjacent water bodies.

Urease-producing bacteria combined with pig manure biochar immobilize Cd and inhibit the absorption of Cd in lettuce (Lactuca sativa L.).

The synergistic remediation of heavy metal-contaminated soil by functional strains and biochar has been widely studied. However, the mechanisms by which urease-producing bacteria combine with pig manure biochar (PMB) to immobilize Cd and inhibit Cd absorption in vegetables are still unclear. In our study, the effects and mechanisms of PMB combined with the urease-producing bacterium TJ6 (TJ6 + PMB) on Cd adsorption were explored. The effects of TJ6 + PMB on the Cd content and pH of the leachate were also studied through a 56-day soil leaching experiment. Moreover, the effects of the complexes on Cd absorption and microbial mechanisms in lettuce were explored through pot experiments. The results showed that PMB provided strain TJ6 with a greater ability to adsorb Cd, inducing the generation of CdS and CdCO3, and thereby reducing the Cd content (71.1%) and increasing the pH and urease activity in the culture medium. TJ6 + PMB improved lettuce dry weight and reduced Cd absorption. These positive effects were likely due to (1) TJ6 + PMB increased the organic matter and NH4+ contents, (2) TJ6 + PMB transformed available Cd into residual Cd and decreased the Cd content in the leachate, and (3) TJ6 + PMB altered the structure of the rhizosphere bacterial and fungal communities in lettuce, increasing the relative abundances of Stachybotrys, Agrocybe, Gaiellales, and Gemmatimonas. These genera can promote plant growth, decompose organic matter, and release phosphorus. Interestingly, the fungal communities were more sensitive to the addition of TJ6 and PMB, which play important roles in the decomposition of organic matter and immobilization of Cd. In conclusion, this study revealed the mechanism by which urease-producing bacteria combined with pig manure biochar immobilize Cd and provided a theoretical basis for safe pig manure return to Cd-polluted farmland. This study also provides technical approaches and bacterial resources for the remediation of heavy metal-contaminated soil.

Soil urease functional stability to Hg pollution: An ecotoxicological perspective.

Mercury (Hg) is a persistent soil pollutant, and its toxicity can be evaluated using soil enzyme indicators. However, a thorough understanding of how the enzyme resists and remains resilient to Hg stress is essential, as it significantly impacts the accuracy of toxicity assessments. Therefore, it is worthwhile to understand the functional stability of urease in soil under Hg pollution. This study compares the effects of Hg at different concentrations and exposure times on soil urease. Results indicate that soil urease activity was enhanced in the first two hours under low levels of Hg pollution, decreased after six hours of acute Hg pollution, and reached its maximum reduction in 24 hours. The urease in fluvo-aquic soil, with higher soil organic matter showed higher resistance to Hg acute pollution than that in red soil. Over a longer aging process, soil urease activity gradually recovered with time. Hormesis effects were observed in red soil under high Hg stress after 30 days, showing the strong resilience of urease enzyme function to Hg pollution. The ecological dose, ED10, (the Hg concentration causing a 10% reduction in soil urease activity) ranged from 0.09 to 0.59 mg kg-1 under short-term exposure, and was lower than that under a longer aging process (0.28 to 2.71 mg kg-1). Further, aging reduced the Hg ecotoxicity due to decreased Hg availability and the resilience of soil urease activity. This indicates that the risk of Hg pollution estimated by soil urease as an indicator depends on exposure time and enzyme stability. These factors need consideration in heavy metal pollution assessments using soil enzymes.

Detection of Helicobacter pylori infection in children using rapid urease and histologic methods of diagnosis.

Objective: The study aimed to detect the presence of Helicobacter pylori infection in children using two investigative methods: the rapid urease test and histological methods. It also examined the relationship between socioeconomic status and Helicobacter pylori infection. Design: This was a cross-sectional study conducted in the paediatric theatre at Korle Bu Teaching Hospital in Accra, Ghana. Participants: Children who were scheduled for upper gastrointestinal endoscopy were recruited into the study. Main outcome measures: The presence of Helicobacter pylori in gastric biopsies was measured using a rapid urease test and histology. Results: Seventy-three children aged 2 years to 16 years were seen during the period. Both tests were positive at the same time in 36 (49.3%) out of the 73 children (p<0.0001). The positivity rates for the rapid urease test and histology were 57.5% and 53.4 %, respectively. Significant predictors of the histology presence of H. pylori were a large household size of at least 6 members (AOR: 4.03; p<0.013) and the presence of pets at home (AOR: 3.23; p<0.044). Conclusions: Substantial agreement was found between the rapid urease test and histology examination of gastric biopsies for the presence of H. pylori. Children from large households and those with pets at home appear to have increased odds of having H. pylori infection of the gastric mucosa. Funding: None declared.

Complete Match of Streptococcus salivarius from Oral Saliva and Stool in a Patient with Hepatic Encephalopathy.

We herein report a 67-year-old Japanese woman with liver cirrhosis caused by primary biliary cholangitis. The patient was admitted to the hospital with loss of consciousness. Hepatic encephalopathy (HE) was diagnosed after diagnostic imaging and symptom assessments. Molecular biology tests were performed on oral saliva and stool samples. The test results indicated sequence similarity between urease-positive S. salivarius in both oral saliva and stool, as revealed by the signals in the overlapping peaks. This bacterium can potentially increase ammonia production in the gut, leading to HE in patients with liver cirrhosis.

Effect and mechanism of Mn2+ on urease activity during anaerobic biological treatment of landfill leachate.

As a crucial hydrolytic enzyme, urease plays a vital role in anaerobic biological treatment. It is well-known that manganese ions are abundant in landfill leachate, but their concentration fluctuates significantly. However, few studies have investigated the effect and mechanism of different concentrations of Mn2+ on urease activity during anaerobic biological treatment of landfill leachate. This paper aimed to investigate the effects and mechanisms of different concentrations of Mn2+ on urease activity. The results showed that an appropriate amount of Mn2+ could significantly enhance urease activity, while a high concentration of Mn2+ could inhibit it. Insight into the mechanisms behind this phenomenon, various methods such as Zeta potential, particle size, ultraviolet spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy, and statistical analysis were employed in our study. Research suggested that, on one hand, Mn2+ may form hydrogen bonds with the side chain amino or carboxyl groups of urease amino acid residues, affecting the structure of urease through hydrogen bonding. Additionally, Mn2+ also binds to urease through hydrophobic interactions. On the other hand, the C-OH and C-N functional groups in urease have a strong affinity for Mn2+, and changes in these functional groups can greatly enhance the activity of urease. Furthermore, under the action of high concentrations of Mn2+, while the structure of urease becomes more stable, there is also a steric hindrance phenomenon that affects the substrate from entering the catalytic center. Therefore, studying the mechanism of Mn2+ affecting urease activity has significant biological significance and provides a new perspective for exploring the impact of metals on anaerobic bioprocessing of landfill leachate.

Host and fungal factors both contribute to cryptococcosis-associated hyperammonemia (cryptammonia).

Cryptococcus neoformans and Cryptococcus gattii are both known urease producers and have the potential to cause hyperammonemia. We hypothesized that the risk of hyperammonemia is increased by renal failure, burden of cryptococcal infection, and fungal strain characteristics. We performed a retrospective review of plasma ammonia levels in patients with cryptococcal infections. Risk factors for hyperammonemia were statistically compared between patients with and without hyperammonemia (>53 µmol/L). Cryptococcal cells from three patients included in the study were recovered from our biorepository. Strain characteristics including urease activity, ammonia production, growth curves, microscopy, melanin production, and M13 molecular typing were analyzed and compared with a wild-type (WT) C. neoformans strain. We included 29 patients, of whom 37.9% had hyperammonemia, 59% had disseminated cryptococcal infection (DCI), and 41% had isolated central nervous system infection. Thirty-eight percent of patients had renal failure and 28% had liver disease. Renal failure was associated with 4.4 times (95% confidence interval [CI] 1.5, 13.0) higher risk of hyperammonemia. This risk was higher in DCIs (RR 6.2, 95% CI 1.0, 40.2) versus isolated cryptococcal meningitis (RR 2.5, 95% CI, 0.40, 16.0). Liver disease and cryptococcal titers were not associated with hyperammonemia. C. neoformans from one patient with extreme hyperammonemia demonstrated a 4- to 5-fold increase in extracellular urease activity, slow growth, enlarged cell size phenotypes, and diminished virulence factors. Hyperammonemia was strongly associated with renal failure in individuals with DCI, surpassing associations with liver failure or cryptococcal titers. However, profound hyperammonemia in one patient was attributable to high levels of urease secretion unique to that cryptococcal strain. Prospective studies are crucial to exploring the significance of this association.IMPORTANCECryptococcus produces and secretes the urease enzyme to facilitate its colonization of the host. Urease breaks down urea into ammonia, overwhelming the liver's detoxification process and leading to hyperammonemia in some hosts. This underrecognized complication exacerbates organ dysfunction alongside the infection. Our study investigated this intricate relationship, uncovering a strong association between the development of hyperammonemia and renal failure in patients with cryptococcal infections, particularly those with disseminated infections. We also explore mechanisms underlying increased urease activity, specifically in strains associated with extreme hyperammonemia. Our discoveries provide a foundation for advancing research into cryptococcal metabolism and identifying therapeutic targets to enhance patient outcomes.

Synergistic biocementation: harnessing Comamonas and Bacillus ureolytic bacteria for enhanced sand stabilization.

Biocementation, driven by ureolytic bacteria and their biochemical activities, has evolved as a powerful technology for soil stabilization, crack repair, and bioremediation. Ureolytic bacteria play a crucial role in calcium carbonate precipitation through their enzymatic activity, hydrolyzing urea to produce carbonate ions and elevate pH, thus creating favorable conditions for the precipitation of calcium carbonate. While extensive research has explored the ability of ureolytic bacteria isolated from natural environments or culture conditions, bacterial synergy is often unexplored or under-reported. In this study, we isolated bacterial strains from the local eutrophic river canal and evaluated their suitability for precipitating calcium carbonate polymorphs. We identified two distinct bacterial isolates with superior urea degradation ability (conductivity method) using partial 16 S rRNA gene sequencing. Molecular identification revealed that they belong to the Comamonas and Bacillus genera. Urea degradation analysis was performed under diverse pH (6,7 and 8) and temperature (15 °C,20 °C,25 °C and 30 °C) ranges, indicating that their ideal pH is 7 and temperature is 30 °C since 95% of the urea was degraded within 96 h. In addition, we investigated these strains individually and in combination, assessing their microbially induced carbonate precipitation (MICP) in silicate fine sand under low (14 ± 0.6 °C) and ideal temperature 30 °C conditions, aiming to optimize bio-mediated soil enhancement. Results indicated that 30 °C was the ideal temperature, and combining bacteria resulted in significant (p ≤ 0.001) superior carbonate precipitation (14-16%) and permeability (> 10- 6 m/s) in comparison to the average range of individual strains. These findings provide valuable insights into the potential of combining ureolytic bacteria for future MICP research on field applications including soil erosion mitigation, soil stabilization, ground improvement, and heavy metal remediation.

Urease-producing bacteria with plant growth-promoting ability that may tolerate and remove cadmium from aqueous solution.

Urease-producing bacteria (UPB) are widely present in soil and play an important role in soil ecosystems. In this study, 65 UPB strains were isolated from cadmium (Cd)-polluted soil around a lead-zinc mine in Yunnan Province, China. The Cd tolerance, removal of Cd from aqueous solution, production of indoleacetic acid (IAA) and plant growth-promoting effects of these materials were investigated. The results indicate that among the 65 UPB strains, four strains with IAA-producing ability were screened and identified as Bacillus thuringiensis W6-11, B. cereus C7-4, Serratia marcescens W11-10, and S. marcescens C5-6. Among the four strains, B. cereus C7-4 had the highest Cd tolerance, median effect concentration (EC50) of 59.94 mg/L. Under Cd 5 mg/L, S. marcescens C5-6 had the highest Cd removal from aqueous solution, up to 69.83%. Under Cd 25 mg/kg, inoculation with B. cereus C7-4 significantly promoted maize growth in a sand pot by increasing the root volume, root surface area, and number of root branches by 22%, 29%, and 20%, respectively, and plant height and biomass by 16% and 36%, respectively, and significantly increasing Cd uptake in the maize roots. Therefore, UPB is a potential resource for enhancing plant adaptability to Cd stress in plants with Cd-polluted habitats.

This study utilized urease-producing bacteria screened from the soil of lead zinc mining areas in Yunnan, China as the research object, enriching the microbial resources in Yunnan. In addition, this article verified the IAA production ability and cadmium removal ability of urease-producing bacteria, and screened out bifunctional urease-producing bacteria that have potential in cadmium pollution control and plant growth promotion.

Synergistic inhibition of ureolytic activity and growth of Klebsiella pneumoniae in vitro suggests cobinding of fluoride and acetohydroxamic acid at the urease active site and provides a novel strategy to combat ureolytic bacteria.

The ability of ureolytic bacteria to break down stable urea to alkaline ammonia leads to several environmental and health challenges. Ureolytic bacteria such as Helicobacter pylori, Klebsiella pneumoniae, and Proteus mirabilis can become pathogenic and cause persistent infections that can be difficult to treat. Inhibiting urease activity can reduce the growth and pathogenicity of ureolytic bacteria. In the present in vitro study, we investigated the synergistic effects of tannic acid (TA) and the urease inhibitors fluoride (F-) and acetohydroxamic acid (AHA). The concentration of AHA needed for efficient inhibition of the ureolytic activity of K. pneumoniae can be significantly reduced if AHA is coapplied with tannic acid and sodium fluoride (NaF). Thus, only 1.20 µmol l-1 AHA in combination with 0.30 mmol l-1 tannic acid and 0.60 mmol l-1 NaF delayed the onset of ureolytic pH increase by 95.8 % and increased the growth lag phase by 124.3 % relative to untreated K. pneumoniae. At these concentrations, without AHA, TA and NaF increased the onset of the ureolytic pH change by only 37.0 % and the growth lag phase by 52.5 %. The strong inhibition obtained with low concentrations of AHA in triple-compound treatments suggests cobinding of F- and AHA at the urease active site and could reduce the side effects of AHA when it is employed as a drug against e.g. urinary tract infections (UTIs) and blocked catheters. This study reports the basis for a promising novel therapeutic strategy to combat infections caused by ureolytic bacteria and the formation of urinary tract stones and crystalline biofilms on catheters.

Chlorogenic acid, caffeic acid and luteolin from dandelion as urease inhibitors: insights into the molecular interactions and inhibition mechanism.

BACKGROUND: Dandelion contains hundreds of active compounds capable of inhibiting urease activity, but the individual compounds have not yet been fully identified, and their effects and underlying mechanisms are not clear. The present study aimed to screen the urease inhibition active compounds of dandelion by urease inhibitory activity evaluation HPLC-tandem mass spectrometry analysis, their mechanism of urease inhibition by polyphenols was explored using enzyme kinetic studies via Lineweaver-Burk plots. Other investigations included isothermal titration calorimetry and surface plasmon resonance sensing, fluorescence quenching experiments, and single ligand molecular docking and two-ligand simultaneous docking techniques. RESULTS: The results indicated that the ethyl acetate fraction of dandelion flower exhibited the greatest inhibition (lowest IC50 0.184 ± 0.007 mg mL-1). Chlorogenic acid, caffeic acid and luteolin could be effective urease inhibitors that acted in a non-competitive inhibition manner. Individually, chlorogenic acid could not only fast bind to urease, but also dissociate rapidly, whereas luteolin might interact with urease with the weakest affinity. The chlorogenic acid-caffeic acid combination exhibited an additive effect in urease inhibition. However, the chlorogenic acid-luteolin and caffeic acid-luteolin combinations exhibited antagonistic effects, with the caffeic acid-luteolin combination showing greater antagonism. CONCLUSION: The present study reveals that chlorogenic acid, caffeic acid and luteolin are major bioactive compounds for urease inhibition, indicating the molecular mechanisms. The antagonistic effects were observed between luteolin and chlorogenic acid/caffeic acid, and the interactions of the catalytic site and flap may account for the antagonistic effects. © 2024 Society of Chemical Industry.

Exploration of morpholine-thiophene hybrid thiosemicarbazones for the treatment of ureolytic bacterial infections via targeting urease enzyme: Synthesis, biochemical screening and computational analysis.

An important component of the pathogenicity of potentially pathogenic bacteria in humans is the urease enzyme. In order to avoid the detrimental impact of ureolytic bacterial infections, the inhibition of urease enzyme appears to be an appealing approach. Therefore, in the current study, morpholine-thiophene hybrid thiosemicarbazone derivatives (5a-i) were designed, synthesized and characterized through FTIR, 1H NMR, 13C NMR spectroscopy and mass spectrometry. A range of substituents including electron-rich, electron-deficient and inductively electron-withdrawing groups on the thiophene ring was successfully tolerated. The synthesized derivatives were evaluated in vitro for their potential to inhibit urease enzyme using the indophenol method. The majority of compounds were noticeably more potent than the conventional inhibitor, thiourea. The lead inhibitor, 2-(1-(5-chlorothiophen-2-yl)ethylidene)-N-(2-morpholinoethyl)hydrazinecarbothioamide (5g) inhibited the urease in an uncompetitive manner with an IC50 value of 3.80 ± 1.9 µM. The findings of the docking studies demonstrated that compound 5g has a strong affinity for the urease active site. Significant docking scores and efficient binding free energies were displayed by the lead inhibitor. Finally, the ADME properties of lead inhibitor (5g) suggested the druglikeness behavior with zero violation.

Measuring Urease and Phospholipase Secretion in Cryptococcus neoformans.

Urease and phospholipase are enzymes that are important virulence factors for Cryptococcus neoformans. These are two of the most studied enzymes involved in how C. neoformans breaches the blood-brain barrier. Additionally, phospholipase secretion also supports dissemination from the lungs. This chapter describes the methods used to measure the secretion of these enzymes, which may be used to characterize strain invasiveness and virulence.

Optimization of immobilized urease enzyme on porous polymer for enhancing the stability, reusability and enzymatic kinetics using response surface methodology.

The study examines the immobilization of the urease enzyme on a range of High Internal Phase Emulsion (polyHIPE) materials, assessing characteristics, efficiency, and performance. It also investigates the impact of polyHIPE type, quantity, incubation time, and various parameters on the process and enzyme activity. Surface morphology and functional groups of polyHIPE materials were determined through scanning electron microscopy (SEM) and fourier transform infrared spectroscopy (FT-IR) analyses, revealing significant alterations after modification with polyglutaraldehyde (PGA). The maximum immobilization efficiency of 95% was achieved by adding PGA to polyHIPE materials with an incubation period of 15 h. The optimized conditions for immobilized enzyme using a Box-Behnken design (BBD) of response surface methodology (RSM) were as follows: temperature (40.8 °C), pH (7.1) and NaCl concentration (0.007 g/L). Furthermore, the immobilized enzyme demonstrated remarkable reusability, retaining 75% of its initial activity after six cycles, and sustained shelf-life stability, retaining over 40% activity after 10 days at room temperature. Kinetic analyses revealed that immobilized urease exhibited higher affinity for the substrate, but lower rate of substrate conversion compared to the free enzyme. These findings offer valuable insights into optimizing urease immobilization processes and enhancing urease stability and activity, with potential applications in various fields, including biotechnology and biocatalysis.

Design and discovery of urease and Helicobacter pylori inhibitors based on benzofuran/benzothiophene-sulfonate and sulfamate scaffolds for the treatment of ureolytic bacterial infections.

A series of sulfonate and sulfamate derivatives bearing benzofuran or benzothiophene scaffold exhibited potent inhibitory effect on urease enzyme. Most of the derivatives exhibited significantly higher potency than thiourea, the standard inhibitor. Compound 1s was identified as the most potent urease inhibitor with an IC50 value of 0.42 ± 0.08 µM, which is 53-fold more potent than thiourea, positive control (IC50 = 22.3 ± 0.031 µM). The docking results further revealed the binding interactions towards the urease active site. Phenotypic screening revealed that compounds 1c, 1d, 1e, 1f, 1j, 1n, and 1t exhibit high potency against H. pylori with MIC values ranging from 0.00625 to 0.05 mM and IC50 values ranging from 0.0031 to 0.0095 mM, much more potent than the positive control, acetohydroxamic acid (MIC and IC50 values were 12.5 and 7.38 mM, respectively). Additional studies were performed to investigate the toxicity of these compounds against the gastric epithelial cell line (AGS) and their selectivity profile against E. coli, and five Lactobacillus species representative of the gut microflora. Permeability characteristics of the most promising derivatives were investigated in Caco-2 cell line. The results indicate that the compounds could be targeted in the GIT only without systemic side effects.

Designing and Synthesis of Novel Fexofenadine-Derived Hydrazone-Schiff bases as Potential Urease Inhibitors: In-Vitro, Molecular Docking and DFT Investigations.

Thirteen novel hydrazone-Schiff bases (3-15) of fexofenadine were succesfully synthesized, structurally deduced and finally assessed their capability to inhibit urease enzyme (in vitro). In the series, six compounds 12 (IC50=10.19±0.16 µM), 11 (IC50=15.05±1.11 µM), 10 (IC50=17.01±1.23 µM), 9 (IC50=17.22±0.81 µM), 13 (IC50=19.31±0.18 µM), and 14 (IC50=19.62±0.21 µM) displayed strong inhibitory action better than the standard thiourea (IC50=21.14±0.24 µM), while the remaining compounds displayed significant to less inhibition. LUMO and HOMO showed the transferring of charges from molecules to biological transfer and MEP map showed the chemically reactive zone appropriate for drug action are calculated using DFT. AIM charges, non-bonding orbitals, and ELF are also computed. The urease protein binding analysis benefited from the docking studies.

Profuse color-evolution based aptasensor for mucin 1 detection utilizing urease-mediated color mixing of the mixed pH indicator.

Mucin 1 is a significant tumor marker, and developing portable and cost-effective methods for its detection is crucial, especially in resource-limited areas. Herein, we developed an innovative approach for mucin 1 detection using a visible multicolor aptasensor. Urease-encapsulated DNA microspheres were used to mediate multicolor change facilitated by the color mixing of the mixed pH indicator, a mixed methyl red and bromocresol green solution. Distinct color changes were exhibited in response to varying mucin 1 concentrations. Notably, the color mixing of the mixed pH indicator was used to display various hues of colors, broadening the range of color variation. And color tonality is much easier to differentiate than color intensity, improving the resolution with naked-eyes. Besides, the variation of color from red to green (a pair of complementary colors) enhanced the color contrast, heightening sensitivity for visual detection. Importantly, the proposed method was successfully applied to detect mucin 1 in real samples, demonstrating a clear differentiation of colors between the samples of healthy individuals and breast cancer patients. The use of a mixed pH indicator as a multichromatic substrate offers the merits of low cost, fast response to pH variation, and plentiful color-evolution. And the incorporation of calcium carbonate microspheres to encapsulate urease ensures stable urease activity and avoids the need for extra urease decoration. The color-mixing dependent strategy opens a new way for multicolor detection of MUC1, characterized by vivid color changes.