STUDIES ON KINETIC PARAMETERS AND BIOCHEMICAL CHARACTERISTICS OF POLYPHENOL OXIDASE PURIFIED FROM JACKFRUIT (ARTOCARPUS HETEROPHYLLUS) WASTE

Objectives: Polyphenol oxidase activity was extensively studied in jackfruit for its role in enzymatic browning. PPO and the phenolic compound play a vital role in defensive mechanism against pest and diseases. Thus, to facilitate further studies in jack fruit waste, Polyphenol oxidase [PPO] was purified and characterized.Methods: Partial Purification of PPO from waste done through a sequential process of ammonium sulfate precipitation, dialysis and ion-exchange chromatography [DEAE- Cellulose]. Then the partially purified PPO was subjected to check various parameters like molecular weight and kinetic activity, the following characteristics of enzyme are checked: SDS-PAGE, pH, temperature, thermal stability, heat inactivation, metal ions, surfactants and inhibitor.Results: Purified PPO resulted in ~23 folds enriched in the specific activity of 1360 [µkat/mg] and it was found to be the monomer with a molecular weight of 63 kDa revealed by Coomasie Brilliant Blue staining. PPO exhibited optimum activity at pH 7.0 and temperature 20oC. PPO showed the maximum stability between pH 6.4- 7.6 at 10 oC - 40 oC. PPO showed the enzyme activity towards Diphenol> Triphenol> Monophenol, the substrate specificity was especially high towards the catechol at 0.1 M. The PPO activity was activated by Mn2+, Triton X- 100, EDTA, Sorbic acid and Citric acid, but inhibited by L- cysteine, Ascorbic acid, SDS, Cetyl trimethyl ammonium bromide [CTAB], K+, Zn2+, Ca2+ and Mg2+. Kinetic constant for PPO was found to be km= 15.82 mM and Vmax= 2182 U/ml min using catechol as substrate.Conclusion: Partial Purification of PPO from waste done through a sequential process of ammonium sulfate precipitation, dialysis and ion-exchange chromatography [DEAE- Cellulose]. The best substrate for PPO was identified as catechol [diphenol] and best inhibitor was L-cysteine and ascorbic acid. Â

Serological and molecular characterization of Cucumber Mosaic Virus (CMV) infecting cucumber (Cucumis sativus L.) in southern Tamil Nadu

Cucumber Mosaic Virus (CMV) is a significant threat to cucumber (Cucumis sativus L.) cultivation, causing mosaic patterns, leaf crinkling, fruit distortion and stunting. This study investigates the serological and molecular characteristics of a CMV- Anantha nambi Nambi kuruchi isolate (CMV-ANK) collected from cucumber plants in major growing areas of Tamil Nadu. Host range studies revealed that CMV-ANK caused systemic infection in multiple crops and chlorotic lesions in okra, chenopodium and wild cucumber. The virus was confirmed using Triple Antibody Sandwich Enzyme-Linked Immunosorbent Assay (TAS-ELISA) and Reverse transcription polymerase chain reaction (RT-PCR) with Coat Protein (CP) gene-specific primers, producing a 1200 bp amplicon. Nucleotide sequencing demonstrated that 99.85 % identity with a CMV isolate from tomato (MZ298672), indicating close genetic similarity. Phylogenetic analysis showed that CMV-ANK clusters with other CMV isolates affecting various crops. Physical property analysis revealed a dilution endpoint at 10-5, thermal inactivation at 65 °C and longevity in vitro up to three days at room temperature and five days under refrigeration. Seed transmission studies confirmed an 83.33 % transmission rate through seeds from infected plants, reinforcing the risk of seed-borne spread. These findings underscore the adaptability and resilience of CMV-ANK, highlighting the need for effective disease management strategies, including the development of resistant cucumber varieties and integrated pest management. The insights gained from this study contribute to understanding CMV epidemiology and offer a foundation for future research aimed at controlling its impact on cucumber production

Identification of Nephelium lappaceum leaves phenolic and flavonoid component with radical scavenging, antidiabetic and antibacterial potential

360-365Nephelium lappaceum Linn. (Rambutan) is traditionally claimed, as a source of natural antioxidants and for its use in the treatment of diabetes and bacterial infections. The present study investigates the in vitro effect of ethanolic Rambutan leaves extract (NL) for its antioxidant effect, α-glucosidase, α-amylase enzyme inhibition, and antibacterial potentials. The total phenolic, total flavonoid content of NL was quantified and were expressed in terms of gallic acid (19.6±0.04 mg GAE/g) and rutin equivalents (16.7±0.01 mg RUE/g) respectively. The antioxidant assay revealed that NL exhibited significant inhibition of DPPH (IC50±SEM: 1.52±0.03 μg/mL) and ABTS (IC50±SEM: 1.295±0.05 μg/mL) radicals. NL also inhibited both α-amylase (IC50±SEM: 2.624±0.07 μg/mL), α-glucosidase (IC50±SEM: 2.416±0.06 μg/mL) enzyme activities, supported by its antioxidant potential and its phenolic and flavonoid content. The antibacterial activity was screened against seven human pathogenic ATCC strains for which the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were recorded. The selected MIC dose was tested, confirmed by Kirby-Bauer agar well diffusion method. NL exhibited MIC and MBC of 62.5 μg/mL and 125 μg/mL against B.subtilis and E.coli respectively. The results support the scientific claim of NL for its antioxidant, antidiabetic and antibacterial potential

Identification of Nephelium lappaceum leaves phenolic and flavonoid component with radical scavenging, antidiabetic and antibacterial potential

360-365Nephelium lappaceum Linn. (Rambutan) is traditionally claimed, as a source of natural antioxidants and for its use in the treatment of diabetes and bacterial infections. The present study investigates the in vitro effect of ethanolic Rambutan leaves extract (NL) for its antioxidant effect, α-glucosidase, α-amylase enzyme inhibition, and antibacterial potentials. The total phenolic, total flavonoid content of NL was quantified and were expressed in terms of gallic acid (19.6±0.04 mg GAE/g) and rutin equivalents (16.7±0.01 mg RUE/g) respectively. The antioxidant assay revealed that NL exhibited significant inhibition of DPPH (IC50±SEM: 1.52±0.03 μg/mL) and ABTS (IC50±SEM: 1.295±0.05 μg/mL) radicals. NL also inhibited both α-amylase (IC50±SEM: 2.624±0.07 μg/mL), α-glucosidase (IC50±SEM: 2.416±0.06 μg/mL) enzyme activities, supported by its antioxidant potential and its phenolic and flavonoid content. The antibacterial activity was screened against seven human pathogenic ATCC strains for which the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were recorded. The selected MIC dose was tested, confirmed by Kirby-Bauer agar well diffusion method. NL exhibited MIC and MBC of 62.5 μg/mL and 125 μg/mL against B.subtilis and E.coli respectively. The results support the scientific claim of NL for its antioxidant, antidiabetic and antibacterial potential

Reliable water quality prediction and parametric analysis using explainable AI models

The consumption of water constitutes the physical health of most of the living species and hence management of its purity and quality is extremely essential as contaminated water has to potential to create adverse health and environmental consequences. This creates the dire necessity to measure, control and monitor the quality of water. The primary contaminant present in water is Total Dissolved Solids (TDS), which is hard to filter out. There are various substances apart from mere solids such as potassium, sodium, chlorides, lead, nitrate, cadmium, arsenic and other pollutants. The proposed work aims to provide the automation of water quality estimation through Artificial Intelligence and uses Explainable Artificial Intelligence (XAI) for the explanation of the most significant parameters contributing towards the potability of water and the estimation of the impurities. XAI has the transparency and justifiability as a white-box model since the Machine Learning (ML) model is black-box and unable to describe the reasoning behind the ML classification. The proposed work uses various ML models such as Logistic Regression, Support Vector Machine (SVM), Gaussian Naive Bayes, Decision Tree (DT) and Random Forest (RF) to classify whether the water is drinkable. The various representations of XAI such as force plot, test patch, summary plot, dependency plot and decision plot generated in SHAPELY explainer explain the significant features, prediction score, feature importance and justification behind the water quality estimation. The RF classifier is selected for the explanation and yields optimum Accuracy and F1-Score of 0.9999, with Precision and Re-call of 0.9997 and 0.998 respectively. Thus, the work is an exploratory analysis of the estimation and management of water quality with indicators associated with their significance. This work is an emerging research at present with a vision of addressing the water quality for the future as well

Studies on heavy metal removal efficiency and antibacterial activity of chitosan prepared from shrimp shell waste

Chitosan, a natural biopolymer composed of a linear polysaccharide of α (1–4)-linked 2-amino 2-deoxy β-d glucopyranose was synthesized by deacetylation of chitin, which is one of the major structural elements, that forms the exoskeleton of crustacean shrimps. The present study was undertaken to prepare chitosan from shrimp shell waste. The physiochemical properties like degree of deacetylation (74.82 %), ash content (2.28 %), and yield (17 %) of prepared chitosan indicated that that shrimp shell waste is a good source of chitosan. Functional property like water-binding capacity (1,136 %) and fat-binding capacity (772 %) of prepared chitosan are in total concurrence with commercially available chitosan. Fourier Transform Infra Red spectrum shows characteristic peaks of amide at 1,629.85 cm(−1) and hydroxyl at 3,450.65 cm(−1). X-ray diffraction pattern was employed to characterize the crystallinity of prepared chitosan and it indicated two characteristic peaks at 10° and 20° at (2θ). Scanning electron microscopy analysis was performed to determine the surface morphology. Heavy metal removal efficiency of prepared chitosan was determined using atomic absorption spectrophotometer. Chitosan was found to be effective in removing metal ions Cu(II), Zn(II), Fe(II) and Cr(IV) from industrial effluent. Antibacterial activity of the prepared chitosan was also determined against Xanthomonas sp. isolated from leaves affected with citrus canker

Integrative analysis of liver-specific non-coding regulatory SNPs associated with the risk of coronary artery disease

Genetic factors underlying coronary artery disease (CAD) have been widely studied using genome-wide association studies (GWASs). However, the functional understanding of the CAD loci has been limited by the fact that a majority of GWAS variants are located within non-coding regions with no functional role. High cholesterol and dysregulation of the liver metabolism such as non-alcoholic fatty liver disease confer an increased risk of CAD. Here, we studied the function of non-coding single-nucleotide polymorphisms in CAD GWAS loci located within liver-specific enhancer elements by identifying their potential target genes using liver cis-eQTL analysis and promoter Capture Hi-C in HepG2 cells. Altogether, 734 target genes were identified of which 121 exhibited correlations to liver-related traits. To identify potentially causal regulatory SNPs, the allele-specific enhancer activity was analyzed by (1) sequence-based computational predictions, (2) quantification of allele-specific transcription factor binding, and (3) STARR-seq massively parallel reporter assay. Altogether, our analysis identified 1,277 unique SNPs that display allele-specific regulatory activity. Among these, susceptibility enhancers near important cholesterol homeostasis genes (APOB, APOC1, APOE, and LIPA) were identified, suggesting that altered gene regulatory activity could represent another way by which genetic variation regulates serum lipoprotein levels. Using CRISPR-based perturbation, we demonstrate how the deletion/activation of a single enhancer leads to changes in the expression of many target genes located in a shared chromatin interaction domain. Our integrative genomics approach represents a comprehensive effort in identifying putative causal regulatory regions and target genes that could predispose to clinical manifestation of CAD by affecting liver function