PHYTOCHEMICAL ANALYSIS AND IN VITRO BIOLOGICAL CHARACTERIZATION OF AQUEOUS AND METHANOLIC EXTRACT OF BACOPA MONNIERI

Objective: The present study was designed to identify the phytocompounds, to compare the antibacterial, antioxidant, and anti-inflammatory effects of aqueous and methanolic extract of Bacopa monnieri.Methods: Antioxidant activity was determined by 1, 1-diphenyl-2-picrylhydrazyl (DPPH), Ferric Reducing Antioxidant Power (FRAP), Super oxide dismutase (SOD), Reduced glutathione (GSH), Catalase assays. Anti-inflammatory activity was measured with inhibition of albumin denaturation and trypsin inhibitory assay. Finally, extracts were tested against various pathogenic bacterial and fungal strains by broth dilution assay and disc diffusion assay respectively.Results: Results showed the presence of alkaloids, flavonoids, phenols, quinines and glycosides etc while steroids and carboxylic acid were absent. The extracts demonstrated free radical-scavenging activity quite comparable with standard ascorbic acid. Methanolic extract exerted comparative higher antioxidant and anti-inflammatory activity than aqueous extract. Both extracts were most effective against Bacillus subtilis and lowest inhibition against Staphylococcus aureus.Conclusion: The results obtained clearly indicated a promising potential of B. monnieri for serving as a strong ROS scavenger, might be used as anti-arthritic and strong natural antibiotic agent for effective treatment of various oxidative stressed disorders (cancer, cardiovascular diseases), inflammatory disorders (rheumatoid arthritis) and various bacterial infections

ASSESSING THE THERAPEUTIC ROLE OF JOSHANDA: PHYTOCHEMICAL, ANTIOXIDANT, ANTI-INFLAMMATORY AND ANTIMICROBIAL ACTIVITIES

Objective: Joshanda, a polyherbal Unani formulation is extensively used as a common home remedy for the treatment of a cough and cold accompanied by pharyngeal inflammation and fever. This study aimed to analyze phytochemicals, antioxidant, anti-inflammatory, and antibacterial activity.Methods: The study investigated the presence of phyto-compounds in joshanda and antioxidant, antibacterial, anti-fungal, and anti-inflammatory activities by various in vitro standard methods using ascorbic acid, ampicillin, and aspirin respectively as standard drugs.Results: Joshanda aqueous extract revealed the presence of tannins, phenols, flavonoids glycosides, terpenoids, and alkaloids and absence of sterols, saponins, xanthoprotiens, and carboxylic acid. Joshanda showed the highest inhibition against B. subtilis (% MGI 99.000±0.577) and least inhibition against P. aeruginosa (%MGI of 84.102±0.491). Joshanda extract, ascorbic acid demonstrated highest % DPPH radical scavenging of 98.379±0.313%, 98.843±0.443% and a minimum of 36.210±1.174%, 83.192±0.422%. Results showed H2O2 scavenging activity of 0.047±0.001 μg/ml per minute degradation of H2O2. FRAP value was observed in joshanda and ascorbic acid with a maximum of 0.945±0.024, 0.687±0.047 mmol and minimum of 0.171±0.036, 0.059±0.005 mmol respectively. Joshanda extract showed the highest albumin denaturation inhibition of 14.069±0.350% and the lowest of 1.880±0.194% at extract volume of 1000 µl and 100 µl respectively. The extract demonstrated the highest proteinase inhibition of 24.003±0.291 % and the lowest of 4.959±0.254% comparable to aspirin. Joshanda had no potent anticandidal activity up to 1 mg/ml.Conclusion: Results clearly suggested that joshanda is a potent phytodrug and can also be used as a strong reactive oxygen species scavenger, might be used as anti-arthritic and strong natural antibiotic agent for effective treatment of various oxidative stressed disorders

Phytochemical analysis and In-vitro Biochemical Characterization of aqueous and methanolic extract of Triphala, a conventional herbal remedy

Triphala, an Indian ayurvedic triherbal formulation, is an equiproportional mixture of fruits of three herbs, amalaki (Emblica officinalis), haritaki (Terminalia chebula) and bibhitaki (Terminalia bellerica). The present study focused on phytocompounds detection and comparative analysis of various biochemical activities in the aqueous and methanolic extracts of triphala and its constituting herbs. Antioxidant activity was determined by 1, 1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), super oxide dismutase (SOD), catalase assay. Antibacterial potential was determined by broth dilution and agar well diffusion assays. Results revealed the presence of valuable bioactive compounds such as flavonoids, alkaloids, phenols, etc which might be responsible for biochemical activities. Extracts exhibited satisfactory radical-scavenging activity comparable with ascorbic acid. Methanolic extracts demonstrated higher antioxidant activity compared to aqueous extract. Extracts showed promising antibacterial potential against tested strain comparable to ampicillin. Hence, it can be concluded that triphala may be a promising candidate in pharmaceuticals and future medicine

Allium sativum Protease Inhibitor: A Novel Kunitz Trypsin Inhibitor from Garlic Is a New Comrade of the Serpin Family.

This study was aimed to purify and characterize the Protease inhibitor (PI) from a plant Allium sativum (garlic) with strong medicinal properties and to explore its phytodrug potentials.Allium sativum Protease Inhibitor (ASPI) was purified using ammonium sulphate fractionation and Fast Protein Liquid Chromatography on anion exchanger Hi-Trap DEAE column. The purified protein was analyzed for its purity and molecular weight by SDS PAGE. The confirmation of presence of trypsin inhibiting PI was performed by MALDI TOF-TOF and analyzed by MASCOT database. The ASPI was further investigated for its kinetic properties and stability under extreme conditions of pH, temperature and chemical denaturants. Secondary structure was determined by Circular Dichorism (CD) spectroscopy.ASPI of ~15 kDa inhibited trypsin and matched "truncated kunitz Trypsin Inhibitor (Glycine max)" in MASCOT database. The purified ASPI showed 30376.1371 U/mg specific activity with a fold purity of 159.92 and yield ~93%. ASPI was quite stable in the range of pH 2-12 showing a decline in the activity around pH 4-5 suggesting that the pI value of the protein as ASPI aggregates in this range. ASPI showed stability to a broad range of temperature (10-80°C) but declined beyond 80°C. Further, detergents, oxidizing agents and reducing agents demonstrated change in ASPI activity under varying concentrations. The kinetic analysis revealed sigmoidal relationship of velocity with substrate concentration with Vmax 240.8 (μM/min) and Km value of 0.12 μM. ASPI showed uncompetitive inhibition with a Ki of 0.08±0.01 nM). The Far UV CD depicted 2.0% α -helices and 51% β -sheets at native pH.To conclude, purified ~15 kDa ASPI exhibited fair stability in wide range of pH and temperature Overall, there was an increase in purification fold with remarkable yield. Chemical modification studies suggested the presence of lysine and tryptophan residues as lead amino acids present in the reactive sites. Therefore, ASPI with trypsin inhibitory property has the potential to be used as a non-cytotoxic clinical agents

A novel multicopper oxidase (laccase) from cyanobacteria: Purification, characterization with potential in the decolorization of anthraquinonic dye - Fig 10

<p>The FTIR spectra (a) Reactive Blue 4 control (b) Laccase treated Reactive Blue 4</p

Protein Estimation and activity profile at each step of purification.

<p>Protein Estimation and activity profile at each step of purification.</p

UV- visible spectra of Reactive Blue-4 after 4 h contact with purified laccase (1.5 U) of <i>Spirulina platensis</i> CFTRI

<p>UV- visible spectra of Reactive Blue-4 after 4 h contact with purified laccase (1.5 U) of <i>Spirulina platensis</i> CFTRI</p

List of peptide fragments obtained after trypsinization (MALDI-TOF/MS-MS analysis).

<p>List of peptide fragments obtained after trypsinization (MALDI-TOF/MS-MS analysis).</p