Maximum Activity Research Articles

Optimization, Purification and Characterization of Lipase from Streptomyces sp. A3301, with Application of Crude Lipase for Cooking Oily Wastewater Treatment

Streptomyces sp. A3301, which produces lipase isolated by Panyachanakul et al. [1]. This optimization was done for the subsequent purification and characterization of the biological lipase produced by the isolate. The results showed that the strain produced lipase with a maximum activity of 321 U/mL using the optimal medium and conditions (1.5 % (w/v) xylose and 2 % (w/v) yeast extract, pH 7.0 at 30 °C, 150 rpm for 3 days). The specific activity of the purified lipase was 27,000 U/mg, which was 544 times the pre-purification level, based on hydrophobic chromatography. After ion-exchange chromatography, the specific activity was 5,600 U/mg, which was 113 times the pre-purification level, with a single-peak purification profile. The purified lipase had a single band based on SDS-PAGE analysis and the molecular mass was 45 kDa. The optimum temperature and thermo-stability of A3301 lipase were 60 and 30 - 55 °C, respectively. The optimum pH of the purified enzyme was pH 9.0, and the enzyme was stable in the pH range of 8.0 - 9.0. The purified lipase was stable in acetone, chloroform and toluene, with a high relative activity of 63 - 76 %. The immobilized strain was then applied to oily-wastewater treatment. The strain can remove oil and grease in synthetic wastewater containing oil at 1 - 3 % (v/v), with removal rates of 100, 99.82 and 99.68 %, respectively, after incubation for 6 days. It was then applied to oily-wastewater treatment from a restaurant, achieving the highest degradation rates of 98.53 % after treatment for 6 days. In addition, it also affected the Biochemical Oxygen Demand (BOD) and Chemical Oxygen Demand (COD) values decreasing. HIGHLIGHTS Optimization of lipase production by Streptomyces A3301. Purification and characterization of the lipase produced by the Streptomyces A3301. Application of the immobilized Streptomyces A3301 strain in the treatment of oily cooking wastewater. The immobilized Streptomyces A3301 strain has the capability to remove oil and grease in both synthetic mediums and restaurant wastewater. GRAPHICAL ABSTRACT

Sumoylation of thymine DNA glycosylase impairs productive binding to substrate sites in DNA

The base excision repair enzyme thymine DNA glycosylase (TDG) protects against mutations by removing thymine or uracil from guanine mispairs and functions in active DNA demethylation by excising 5-formylcytosine (fC) and 5-carboxylcytosine (caC). Post-translational modification of TDG by SUMO (small ubiquitin-like modifier) reduces its glycosylase activity but the mechanism remains unclear. We investigated this problem using biochemical and biophysical approaches and a TDG construct comprising residues 82-340 (of 410) that includes the SUMOylation site and the motif for non-covalent SUMO binding. Single turnover kinetics experiments were collected at multiple enzyme concentrations ([E]) and the hyperbolic dependence of activity (kobs) on [E] yielded the maximal glycosylase activity (kmax), the enzyme concentration giving half-maximal activity (K0.5), and the catalytic efficiency (kmax/K0.5). Sumoylation of TDG (or TDG82-340) causes large reductions in catalytic efficiency for G·T, G·U, G·fC, and G·caC DNA substrates, due largely to weakened substrate affinity (increased K0.5). 19F NMR experiments show that sumoylation of TDG82-340 reduces productive binding to G·U mispairs and dramatically impairs binding to G·T mispairs. A mutation in the TDG SUMO-interacting motif (SIM), E310Q, shown previously to perturb noncovalent binding of SUMO to unmodified TDG, rescues the glycosylase activity of sumoylated TDG82-340. Similarly, NMR studies show the mutation restores productive binding of sumoylated TDG82-340 to G·U and G·T pairs. Together, the results indicate that intramolecular SUMO-SIM interactions mediate the adverse effect of sumoylation on TDG activity and suggest a model whereby the disruption of SUMO-SIM interactions enables productive binding of sumoylated TDG to substrate sites in DNA.

Effects of altered contractile environment on muscle shape change in the triceps surae.

Skeletal muscles change shape when they contract. Current insights into the effects of shape change on muscle function have primarily come from experiments on isolated muscles operating at maximal activation levels. However, when muscles contract and change shape, the forces they apply onto surrounding muscles will also change. The impact of an altered contractile environment (i.e., mechanical behaviour of surrounding muscle) on muscle shape change, remains unknown. To address this, we altered the mechanical contributions of the gastrocnemii during isometric plantarflexion contractions [via changing knee angle] and determined if there were associated changes in how the muscles of the triceps surae bulged in thickness during a ramped contraction. We combined B-mode ultrasound imaging with surface electromyography to quantify the neuromechanical contributions of the medial (MG) and lateral gastrocnemius (MG) and soleus (SOL) muscles during isometric plantarflexion contractions. Our results demonstrated that at the same SOL activity levels, altering knee angle had no influence on the magnitude of muscle shape change (thickness) in the triceps surae muscles. We observed high levels of inter-individual variability in muscle bulging patterns, particularly in the knee flexed position, suggesting a complex relationship between muscle bulging and activation strategies in the triceps surae, which may be related to differences in muscle mechanical properties between participants or across muscles. Our findings highlight the dynamics of in vivo bulging interactions among muscles within the triceps surae and provide insights for future investigations into the impact of altered contractile environments on three-dimensional muscle deformations and force production.

Assessing Algerian Ilex aquifolium L.’s phenolic compounds, antioxidant and enzyme inhibitory properties to find a new natural remedy for Alzheimer’s, diabetes and skin diseases

This research explored the impact of methanolic extracts from the fruits and leaves of Ilex aquifolium L., a rare Algerian species, on several enzymes, focusing on tyrosinase, α-amylase, α-glucosidase, and cholinesterases, as well as their antioxidant potential in vitro assays, including phosphomolybdenum, DPPH, ABTS, CUPRAC, FRAP, and MCA. HPLC-DAD analysis identified chlorogenic acid, rosmarinic acid, hesperidin, and quercetin as predominant phenolic components. Both extracts exhibited notable antioxidant properties, up to 250.08 mg TE/g for fruits and 237.08 mg TE/g for leaf extracts. Enzyme inhibitory assays demonstrated significant inhibition against AChE and BChE with maximum activities of 2.34 mg GALAE/g in leaves and 4.32 mg GALAE/g in fruits. Additionally, considerable tyrosinase inhibition was observed in leaves at 39.69 mg KAE/g. These findings highlight I. aquifolium L. as a valuable source for investigating natural antioxidants and enzyme inhibitors, particularly for possible cosmetics and food additives applications.

Characterization of a Peptidoglycan-Degrading Protein: Biochemical and Antimicrobial Characteristics, Antibiotic Synergism, and Delivery System Innovation.

The global health threat posed by antibiotic resistance has led to new research involving bacteriophage-encoded enzymes. This study characterized a new peptidoglycan-degrading protein and evaluated its synergism with colistin and its antimicrobial efficacy when conjugated with polycationic-polymer nanoparticles. The gene that codes for endolysin in the vB_PaeM_USP2, a Pseudomonas aeruginosa bacteriophage, was cloned and expressed in Escherichia coli. The recombinant endolysin (rEnd2) was purified and its biochemical properties were determined using peptidoglycan substrate. The enzymatic activity was measured through peptidoglycan layer degradation and a decrease in turbidity of permeabilized Gram-negative bacteria. The antimicrobial activity of rEnd2, alone and in combination with colistin, was evaluated by checkerboard assay. The antibacterial activity of the cationic lipid oleylamine (OAM) conjugated with rEnd2 (OAM-rEnd2) was evaluated by time killing assay. The rEnd2 is structurally analogue with other endolysins and showed muramidase activity. The rEnd2 maintained higher activity between pH 6.0 to 7.5, had maximum activity at 35 °C, and was not affected by chaotropic and reducing reagents. It was sensitive to an increase in surfactant concentration, being inactivated by sodium dodecyl sulfate and cetyltrimethylammonium bromide. Ions exhibited neither a positive nor a negative effect on enzyme activity. The rEnd2 showed clear muralytic activity and decreased turbidity of permeabilized Gram-negative bacteria. However, it did not control bacterial growth despite the combination with an antibiotic and its complexation with polycation (OAM-rEnd2 nanoparticle conjugate). The rEnd2 did not show clear antimicrobial activity suggesting further optimization of conditions for its activity or engineering and modification.

Cost-Effective Optimization of the Transfructosylation Activity of an Invertase Produced from Aspergillus carbonarius PC-4 Using Pineapple Crown and Determination of Its Biochemical Properties

Fructooligosaccharides are prebiotic sugars that are widely used in the production of functional foods, which can be produced enzymatically by the transfructosylation reaction of sucrose. This work aimed to optimize the production of an invertase with high transfructosylation activity from Aspergillus carbonarius PC-4 using pineapple crown as the inducer substrate and evaluate its biochemical properties. The culture medium was optimized using a Plackett–Burman experimental design and a central composite rotatable design, resulting in a maximum transfructosylation activity of 65.33 U/mL at 72 h of cultivation. The cultivation parameters were Yp/s = 1070.75 U/g and PP = 2771.48 U/h, which showed an increase of 5.2-fold in the enzyme produced. The optimum temperature (50 °C) and pH (5.0) for the enzymatic activity were obtained by a CCR design. The enzyme showed a half-life of 60 min at 40 °C. In conclusion, the invertase produced from A. carbonarius PC-4 using agro-industrial waste (pineapple crown) and an inorganic nitrogen source (ammonium nitrate) exhibits high transfructosylation activity that can be used as a potential source for the production of fructooligosaccharides.

Isolation of a novel Bacillus strain with industrial potential of producing alkaline chitosanase

A novel Bacillus strain, designated as HZ20-1, was discovered. This strain can produce naturally alkaline chitosanase without induction. Genomic analysis revealed that this chitosanase belongs to glycoside hydrolase family 8. When colloidal chitosan is used as a substrate, the maximum enzyme activity of 2.39 ± 0.03 U/mL is observed at a pH range of 8.5–9. This alkaline enzyme holds advantages over common acidic enzymes in various fields such as medicine, the environment, and daily chemical products, and thus has great market value. Moreover, as the chitosanase is a constitutive enzyme, there is no need for substrate induction. This can simplify fermentation equipment and reduce production cost. Additionally, in a preliminary study, we found that this strain can also degrade chitin and chitosan derivatives. After analysis, it was discovered that it has genes in glycoside hydrolase families 18 and 23. By controlling the enzymatic hydrolysis time, it is possible to produce products with different molecular weights, including N-acetylglucosamine, glucosamine, chito-oligosaccharides, and chitin oligosaccharides. Consequently, Bacillus sp. HZ20-1 is considered to have great potential for future industrial production of enzymes and oligosaccharides.

Enhanced biochemical properties of soybean root nodule asparaginase through plant molecular farming compared to bacterial enzyme for cancer treatment

Asparaginase is a therapeutic enzyme used as an anticancer agent and is typically produced through microbial fermentation using organisms such as Escherichia coli and Erwinia chrysanthemi. However, this method faces challenges, including potential enzyme contamination during production, allergic reactions to the enzyme, and stability issues requiring stringent control measures. An innovative solution is the application of plant molecular farming, utilizing Rhizobium root symbiosis for asparaginase production. The objective is to optimize nodule development for asparaginase yield, characterize the enzyme's properties, and evaluate its anticancer efficacy against microbial enzyme. In our study, we established soybean root cultures and inoculated them with Bradyrhizobium japonicum to form root nodules. We evaluated eukaryotic asparaginase production at different incubation times. We purified asparaginase from the root nodule cultures and compared its physicochemical properties and anticancer activity with microbial asparaginase. Results showed that asparaginase reached maximum activity in root nodule cultures 10 days after rhizobium inoculation in the culture media. The root nodule asparaginase exhibited a high content of alpha helices and beta sheets and a low random coil. It demonstrated higher stability and activity across different pH levels and temperatures than Escherichia coli asparaginase. Additionally, root nodule asparaginase displayed better catalytic parameters and stability over time than E. coli asparaginase. Thus, root nodule asparaginase is superior to E. coli asparaginase as an anticancer agent. This ensures the root nodule asparaginase can effectively target cancer cells, enhancing the overall therapeutic outcome. This provides a renewable, cost-effective, and environmentally friendly alternative to traditional enzyme production methods.

Glial-modulating agents for the treatment of pain: a systematic review.

Preclinical research supports a critical role for nervous system glia in pain pathophysiology. This systematic review of human trials of potential glia-modulating drugs for the prevention or treatment of pain followed a predefined search strategy and protocol registration. We searched for English language, randomized, double-blind trials comparing putative glia-modulating drugs to placebo or other comparators. The primary outcomes included validated participant-reported measures of pain intensity or relief and, in studies of opioid administration, measures of opioid consumption and/or opioid-related adverse effects. Twenty-six trials (2132 participants) of glial modulators (12 minocycline, 11 pentoxifylline, and 3 ibudilast) were included. Because of clinical heterogeneity related to study drug, participant population, outcome measures, and trial design, no meta-analysis was possible. Only 6 trials reported a positive effect of the treatment (pentoxifylline-4 trials; minocycline-2 trials), whereas 11 trials reported mixed results and 9 trials reported no effect. This review does not provide convincing evidence of efficacy of current pharmacological targets of nervous system glial function for pain treatment or prevention. However, in light of ample preclinical evidence of the importance of neuroimmune signalling and glial functions in pain pathophysiology, continued strategic human research is anticipated to identify (1) drugs with maximal activity as selectively targeted glial modulators, (2) the necessary timing and duration of pharmacological glial modulation needed for pain prevention or treatment for specific injuries or pain conditions, and (3) the best design of future clinical trials of glial-targeted drugs for pain treatment and/or prevention.

Improvement of Thermo-Stability and Solvent Tolerant Property of Streptomyces sp. A3301 Lipase by Immobilization Techniques with Application in Poly (lactic acid) Polymerization by Using Biological Process

The thermo-solvent-tolerant lipase-producing actinomycete, Streptomyces sp. A3301, was utilized as a biocatalyst for poly (lactic acid) or PLA polymerization. The study aimed to optimize lipase immobilization conditions, characterize the immobilized lipase and apply it for PLA polymerization. The results showed using a sponge as the immobilizing matrix was the most effective method, achieving a maximum activity of 277 U/g of sponge. The optimal sponge size was determined to be 0.125 cm³ and pre-soaking the sponge in 0.1 M phosphate buffer at pH 7.0 for 24 h before use proved advantageous. Immobilization significantly enhanced the thermo-stability of the enzyme, with a relative activity ranging from 140 to 190 % within the temperature range of 30 to 60 °C. In contrast, the crude lipase exhibited thermo-stability only within the 30 - 50 °C range. The immobilized lipase demonstrated stability under PLA polymerization conditions, which involved a reaction mixture containing toluene and lactic acid and performed at 60 °C for 8 h. The immobilized lipase maintained its activity under this condition for 5 h, retaining a relative activity of 230 %, which was 1.2 times higher than the activity of the crude lipase. When the immobilized lipase was used in PLA polymerization, the resulting PLA product exhibited a molecular weight of 5,333 ± 0.02 Da, and the degree of polymerization was approximately 72. These findings underscore the potential of the immobilization technique to enhance lipase activity for PLA polymerization. HIGHLIGHTS PLA polymerization via a biological process is demonstrated. PLA can be polymerized by enzyme lipase produced by Streptomyces sp. A3301. Improving PLA polymerization with enzyme immobilization techniques. Sponge is the best immobilizer for PLA polymerization. PLA can be synthesized under mild conditions. GRAPHICAL ABSTRACT

Clinical value of SUVpeak-to-tumor centroid distance on FDG PET/CT for predicting neoadjuvant chemotherapy response in patients with breast cancer

BackgroundSince it has been found that the maximum metabolic activity of a cancer lesion shifts toward the lesion edge during cancer progression, normalized distances from the hot spot of radiotracer uptake to tumor centroid (NHOC) and tumor perimeter (NHOP) have been suggested as novel F-18 fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) parameters that can reflect cancer aggressiveness. This study aimed to investigate whether NHOC and NHOP parameters could predict pathological response to neoadjuvant chemotherapy (NAC) and progression-free survival (PFS) in breast cancer patients.MethodsThis study retrospectively enrolled 135 female patients with breast cancer who underwent pretreatment FDG PET/CT and received NAC and subsequent surgical resection. From PET/CT images, normalized distances of maximum SUV and peak SUV-to-tumor centroid (NHOCmax and NHOCpeak) and -to-tumor perimeter (NHOPmax and NHOPpeak) were measured, in addition to conventional PET/CT parameters.ResultsOf 135 patients, 32 (23.7%) achieved pathological complete response (pCR), and 34 (25.2%) had events during follow-up. In the receiver operating characteristic (ROC) curve analysis, NHOCmax showed the highest area under the ROC curve value (0.710) for predicting pCR, followed by NHOCpeak (0.694). In the multivariate logistic regression analysis, NHOCmax, NHOCpeak, and NHOPmax were independent predictors for pCR (p < 0.05). In the multivariate survival analysis, NHOCpeak (p = 0.026) was an independent predictor for PFS along with metabolic tumor volume, with patients having higher NHOCpeak showing worse PFS.ConclusionNHOCpeak on pretreatment FDG PET/CT could be a potential imaging parameter for predicting NAC response and survival in patients with breast cancer.

Discovery of 5-(Substituted Phenyl)-2-aryl Benzimidazole Derivatives as SIRT1 Activators: Their Design, in silico Studies, Synthesis, and in vitro Evaluation.

Silent information regulator two homologue one (SIRT1) is an emerging target for managing metabolic disorders. This study aimed to synthesize novel 5-(- substituted phenyl)-2-aryl benzimidazole derivatives and evaluate them for SIRT1 activation. The compounds were designed according to the findings of the QSAR models framed in our previous studies. Molecular docking and dynamics studies were also performed to explore the interactions of designed compounds with the active site of the SIRT1 enzyme using AutoDock Vina and Schrödinger Maestro version 11.8.012, respectively. Compounds with good binding affinity were synthesized by Suzuki-Miyaura cross-coupling and spectrally characterized. The molecules were evaluated for their in vitro SIRT1 activation properties using a fluorescent screening kit. Based on the results of in vitro assay, a structure-activity relationship was established. SwissADME was employed to calculate the pharmacokinetics characteristics of the synthesized molecules. The molecular docking studies revealed that all the activators were effectively docked in the catalytic active site. All compounds demonstrated interactions with important amino acids like Glu230 and Arg446. In molecular dynamics simulations, the root mean square deviation (RMSD) of compound 5m and protein SIRT1 remained stable, i.e., below 3mm. Compound 5m, 4-(2-(3,4-dihydroxy-5-nitrophenyl)-1H-benzo[d]imidazol- 5-yl)benzaldehyde, was the most potent compound with an EC50 value of 0.006 mM (±0.001) and maximum activation of 240.5%. All the synthesized compounds had acceptable theoretical ADME profiles, and drug-likeness properties complied with Lipinski's rule. According to the findings, synthesized compounds may be viable leads for SIRT1 activators and may be used to advance preclinical in vivo research utilizing animal models.

A study method using early dynamic acquisition of [18F]fluorodopa positron emission tomography for the differential diagnosis between progression and radionecrosis of brain metastases after radiotherapy

BackgroundIt is difficult to distinguish between the brain metastasis progression (BMP) and brain radionecrosis (BRN) on the basis of 18F-3,4-dihydroxyphenylalanine positron emission tomography/computed-tomography (18F-FDOPA PET/CT) data. The advent of silicon photomultiplier (SiPM) PET technology makes it possible to study dynamic volumes and potentially improve diagnostic accuracy. We developed a method for processing 18F-FDOPA PET/CT in the differential diagnosis between BMP and BRN. The method involves a short (3-second) sampling time during a 4-minute acquisition on a SiPM-PET/CT machine. We prospectively included 15 patients and 19 metastases. All acquisitions were performed in list mode acquisition for 25 min on a four-ring SiPM PET/CT system. We calculated the ratios between the maximum activity in the lesion’s voxel and the mean activity in the contralateral region (VOImax/CLmean) or the mean activity in the white matter (VOImax/WMmean).ResultsSeven lesions were classified as BMP and twelve were classified as BRN. Statistically significant intergroup differences in the VOImax/CLmean and VOImax/WMmean activity ratios were observed for both the clinical volume and the early acquisition. The best performing quantitative variable was the VOImax/CLmean ratio on early acquisition, with a diagnostic accuracy of 94.7%, a sensitivity of 100%, and a specificity of 91.7%.ConclusionThe 18F-FDOPA PET/CT data acquired a few minutes after the bolus injection confirms its value in differentiating between BMP and BRN, compared to the much longer classic clinical protocol.

Production of invertase from Penicillium spp. under solid state fermentation and its immobilization on magnetic nanoparticles

Invertase is an industrially significant enzyme that catalyzes the hydrolysis of sucrose into glucose and fructose. The present work aimed to produce invertase from Penicillium spp. under solid-state fermentation, utilizing banana peel waste as the substrate, and to characterize and immobilize the enzyme using chitosan-coated magnetic nanoparticles. The partially purified invertase was obtained via ammonium sulfate precipitation. The stability of crude, partially purified, and immobilized invertase was assessed for various parameters such as pH, temperature, thermal stability, metal ions, and chemical inhibitors. Invert syrup was produced via continuous bioconversion. The optimal incubation period for invertase production was observed to be four days, resulting in a maximum invertase activity of 75 U/ml. The maximum activities of the crude and partially purified invertase were observed at pH 6, temperature 50°C, and metal ion KCl, while the chemical inhibitor EDTA enhanced the activity. Partial purification of the crude invertase with 70% ammonium sulfate yielded 49 U/ml, and immobilization on chitosan-coated magnetic nanoparticles resulted in a 59.01% immobilized yield. The maximum immobilized invertase activity was observed at 70°C, and the immobilization procedure did not significantly influence pH, metal ions, or chemical inhibitor activity. Invert syrup production via continuous bioconversion resulted in an enzyme activity of 31 U/ml. This study highlights the potential applications of invertase in the food industry, pharmaceuticals, confectionaries, and related fields.

Topology-defined bioactive properties of porous Ti-6Al-4V scaffolds produced by laser powder bed fusion

The effect of the design of additively printed Ti porous structures on their bioactive properties is elucidated by in vitro testing. 3D-printed scaffolds with pores defined by several bioinspired triply periodic minimal surfaces as well as by a strut model have been engineered. Cell proliferation, angiogenesis and osteogenesis have been studied with the help of multipotent mesenchymal stromal cells. The elementary unit size and pore design can notably modify bioactive properties of porous structures. The models providing maximal bioactive activity of developed scaffolds are discussed.

Enhanced biocatalytic laccase production using agricultural waste in solid-state fermentation by Aspergillus oryzae for p-chlorophenol degradation

Agricultural residues are one of the most cost-effective and readily accessible carbon resources for producing commercially significant enzymes. Several enzymes have been used in different industries like pharmaceuticals, foods, textiles, and dyes that can be generated by various species of microbes found in waste from agriculture. The current research investigated laccase production by Aspergillus oryzae utilizing agricultural wastes. Physical and chemical properties, including pH, temperature, sucrose, yeast extract, and copper sulfate levels, were optimized. The utilization of the response surface methodology along with the centralized composite design method, which assesses multiple media parameters and utilizes a two-level experimental approach, aids in determining the variable and its significance in increasing production quality. The centralized composite design enhancement showed that the optimal conditions for highest laccase activity (623.16 U/mL) were pH 7.0, temperature 25 °C, corn cobs as substrate, sucrose (2.0 %), yeast extract (1.0 %), and copper sulfate (0.1 mM) level. The laccase enzyme was optimized using various pH, temperature, metal ions, and inhibitors combinations. The extracted laccase enzyme maximum activity was attained at pH 6.0 and 40 °C. The inclusion of divalent ions can enhance laccase activity, while the use of various inhibitors decreases laccase activity. Under various pH circumstances, the Aspergillus oryzae laccase enzyme can successfully degrade p-chlorophenol. The present study describes statistically validated optimal methodologies for enhancing laccase synthesis, leading to a laccase production technique that is simultaneously highly efficient and economically profitable, with possible use of p-chlorophenol degradation.

Exploring Phytochemical Profile and Biological Activities of Callistemon viminalis (Sol. ex Gaertn.) G.Don ex Loudon) By In vitro and In silico Approaches.

Callistemon viminalis (Sol. ex Gaertn.) G.Don ex Loudon) (family: Myrtaceae) is used for its medicinal properties in treating various metabolic disorders. We investigated the chemical characterization and biological screening of the n-hexane extract of C. viminalis. The total phenolic content was (37.45±7.40 mg GA.E/g D.E±S.D) and the total flavonoid content was (18.43±6.34 mg R.E/g D.E±S.D). GC-MS screening of the n-hexane extract tentatively identified 70 bioactive phytochemicals. The maximum antioxidant potential (289.99±9.01 mg T.E/g D.E±S.D) was observed via the FRAP assay. Enzyme inhibition assays revealed that n-hexane extract of C. viminalis showed enzyme inhibition against the enzymes including α-glucosidase (6.9±0.13 mmol of ACA.E/g D.E±S.D); α-amylase (7.2±0.56 mmol of ACA.E/g D.E±S.D), urease (4.95±0.9 mg of TU.E/g D.E±S.D), acetylcholinesterase (2.9±0.08 mg GALA.E/g D.E±S.D), lipoxygenase (4.93±1.05 mg of Indo.E/g D.E±S.D) and tyrosinase (4.33±0.62 mg of KA.E/g D.E±S.D). The extract showed maximum antibacterial activity against Staphylococcus aureus (71.30±4.44 %) followed by Bacillus subtilis (68.55±2.70 %), Pseudomonas aeruginosa (57.86±6.02 %), and Salmonella typhi (53.90±5.05 %). Docking studies revealed good docking interactions between ligands and the studied enzymes, whereas ADME analysis revealed the pharmacokinetic profiles of the phytoconstituents. C. viminalis possesses promising therapeutic potential and can be further explored for drug development and drug design.

Screening and Optimization of Pathogenic Strains of Pseudomonas sp. for Cholinesterase Enzyme Activity

The identification and optimization of pathogenic strains of Pseudomonas sp. with significant Cholinesterase enzyme activity has promising implications for environmental and medical applications. This study aimed to screen various Pseudomonas strains to identify those exhibiting high cholinesterase activity and to optimize the conditions for maximal enzyme production. Multiple Pseudomonas strains were isolated and characterized from diverse clinical samples, employing selective media and biochemical assays to determine cholinesterase activity. The strains with maximum enzyme activity were further subjected to optimization experiments, including variations in growth conditions such as Temperature, pH and Nutrient availability. The findings revealed sample no.39 among the collected 50 samples had significant enzyme activity of 0.19151 IU is obtained when choline chloride was 0.3999%, Ammonium Chloride was 0.01M, pH was 7.0, Temperature was 27°C and Time of incubation was 24hrs. These results not only advance our understanding of Pseudomonas sp. enzyme capabilities but also offer potential applications in bioremediation, biosensor development and pharmaceutical industries. Future research will focus on the molecular characterization of the cholinesterase enzyme and its applications in various industrial processes.

Effect of Different Kharif Paddy Straw Management Options and Different Levels of Nitrogen on Soil Dehydrogenase and Urease Activity

A field experiment was conducted during rabi 2020-21, at Regional Agricultural Research Station, Polasa, Jagtial, PJTSAU to study soil dehydrogenase and urease activity as influenced by different kharif paddy straw management options and nitrogen levels. The results reveal that dehydrogenase and urease activity of soil was significantly influenced by paddy straw management options and activity was increased with crop duration up to 60 DAT and decreased after harvest. Dehydrogenase activity was reduced after burning (1.93 mg TPF produced g-1 soil d-1) as compared to the initial values (2.34 mg TPF produced g-1 soil d-1). The highest dehydrogenase activity of 3.74 and 3.54 mg TPF produced g-1 soil d-1 was recorded at 60 DAT in paddy straw incorporation with phosphorus and incorporation without phosphorus and decreased to 3.36 and 3.03 mg TPF produced g-1 soil d-1 after harvest respectively. Soil urease activity was showed an increasing trend with crop duration up to 60 DAT and then declined. Maximum activity of 3.05, 4.84, 5.17 mg NH4+ released g-1 soil h-1 was observed at 60 DAT under paddy straw burning, paddy straw incorporation with phosphorus incorporation without phosphorus respectively. Paddy straw incorporation with phosphorus recorded higher urease activity and paddy straw burning recorded lower urease activity at all the intervals of crop growth. The effect of nitrogen levels on soil urease activity was found to be significant. Soil urease was highest with 120% RDN followed by 115% RDN, followed by 110% RDN, followed by 100% RDN and the similar trend was observed at all the intervals of crop growth.

Optimization of Cellulase Production from Agri-Industrial Residues by Aspergillus terreus NIH2624

The objective of this study was to assess the cellulase production of four fungi: Aspergillus terreus NIH2624, Aspergillus clavatus NRRL1, Aspergillus versicolor CBS583.65 and Aspergillus phoenicis ATCC3157, under submerged cultivation conditions. When these fungi were cultured in shake flasks using Mandels and Weber’s minimal medium with 1% sugarcane bagasse as a carbon source and 1.8 g/L of rice bran extract as a nitrogen source, A. terreus showed maximum cellulase production (filter paper activity (FPase) 3.35 U/mL; carboxymethyl cellulase activity (CMCase) 1.69 U/mL). Consequently, A. terreus was selected for the optimization study for cellulase production. Among the different tested carbon sources, A. terreus showed higher CMCase activity when it was cultivated on delignified sugarcane bagasse (1.64 U/mL) and higher FPase activity on sugarcane straw (7.95 U/mL). Regarding the nitrogen sources, the maximum FPase activity was observed when using rice bran (FPase, 8.90 U/mL) and soybean meal (FPase, 9.63 U/mL). The optimized fermentation medium (minimal medium with delignified sugarcane bagasse and rice bran as carbon and nitrogen sources, respectively) resulted in an enzymatic cocktail mainly composed of xylanases, with a maximum activity of 1701.85 U/mL for beechwood xylan, 77.12 U/mL for endoglucanase and 21.02 U/mL for cellobiohydrolase. Additionally, the enzymatic cocktail showed efficient activities for β-glucosidase, β-xylanase, arabinofuranosidase and lytic polysaccharide monoxygenases (LPMOs). This cellulase enzyme solution has the potential to efficiently hydrolyze lignocellulosic biomass, producing second-generation sugars in biorefineries.