Optimization of production, biochemical characterization and In Vitro evaluation of the therapeutic potential of fibrinolytic enzymes from a new Bacillus Amyloliquefaciens

The capacity of fibrinolytic enzymes to degrade blood clots makes them of high relevance in medicine and in the pharmaceutical industry. In this work, forty-three microorganisms of the genus Bacillus were evaluated for their potential to produce fibrinolytic proteases. Thirty bacteria were confirmed as producers of fibrinolytic enzymes, the best results obtained for the strain Bacillus amyloliquefaciens UFPEDA 485. The optimization of the enzyme production conditions was done by a central composite design (CCD) star 23 that allowed to define the optimal conditions for soybean flour and glucose concentrations and agitation rate. The highest fibrinolytic activity (FA) of 813 U mL-1 and a degradation of blood clot in vitro of 62% were obtained in a medium with 2% (w/v) of soybean flour and 1% (w/v) glucose at 200 rpm after 48 h of cultivation, at pH 7.2 and 37 °C. The obtained fibrinolytic enzyme was characterized biochemically. Fibrinolytic activity was inhibited by PMSF (fluoride methylphenylsulfonyl - C7H7FO2S) 91.52% and EDTA (ethylenediaminetetraacetic acid - C10H16N2O8) 89.4%, confirming to be a serine- metallo protease. The optimum pH and temperature were 7.0 and 37 oC, respectively, and the enzyme was stable for 12 h. The fibrinolytic activity at physiological conditions of this enzyme produced by Bacillus amyloliquefaciens UFPEDA 485, as well as its long term stability, demonstrate that it has suitable characteristics for human and veterinary applications, and promises to be a powerful drug for the treatment of vascular diseases.We express our thanks to Coordination for the Improvement of Higher Level Education Personnel (CAPES) - Doctoral Sandwich Program (PDSE) Nº 0259/ 12-8 and National Council for Scientific and Technological Development (CNPq) - Nº 202026/2011-6 for the financial support

Antimicrobial, Antioxidant and Cytotoxic Activity of MarineStreptomyces parvulus VITJS11 Crude Extract

The main aim of the study was to evaluate the bioactive properties of ethyl acetate crude extract of Streptomyces parvulus VITJS11 with a view to assess their therapeutic potential. The biological activity of ethyl acetate extract was tested against fungal and bacterial pathogens. The free radical scavenging potential of the crude extract was determined by DPPH assay. The chemo preventive properties of S. parvulus VITJS11 ethyl acetate extract was examined by MTT assay on HepG2 cells. The morphological, physiological and the biochemical properties of the strain S. parvulus VITJS11 was confirmed by conventional methods. Genotypic characterization was done using 16S r-DNA partial gene amplification and sequencing. The authenticity of the crude chemical constitutes were determined by the GC-MS. The ethyl acetate extract of VITJS11 showed maximum antifungal activity against three Aspergillus species and prominent antibacterial activity against two Gram positive and Gram negative bacteria at 20 mg/mL. The antioxidant potential of the crude extract exhibited strong reducing power activity at 5mg/ mL with 85% inhibition and the cytotoxic effect was found with IC50 of 500µg/ mL on HepG2 cell lines. The GC-MS analysis and the chromatogram patterns revealed 16 peaks, indicating the presence of bioactive constituents, which included several important organic compounds, namely 9-(2',2'-dimethylpropanoilhydrazono)-3,6-dichloro-2,7-Bis-[2-(diethylamino)-ethoxy]fluorine (23.1) Dotriacontylpentafluoropropionate,(25.0) Octadecanoic acid, (20.0); Trans-2-methyl-4-n-butylthiane, S, S-dioxide.(19.0). The results showed the benefit of ethyl acetate extract from S. parvulus VITJS11 in treating microbial infections and indicated their broad spectrum of activity with beneficial virtues for therapeutic use

BRAZILIAN ARCHIVES OF BIOLOGY AND TECHNOLOGY A N I N T E R N A T I O N A L J O U R N A L Antimicrobial, Antioxidant and Cytotoxic Activity of Marine Streptomyces parvulus VITJS11 Crude Extract

ABSTRAC

A comparative study of the lipase yield by solid state and submerged fermentations using fungal species from biopharmaceutical oil waste

A comparative study of lipase enzyme yields by solid state fermentation (SSF) and submerged fermentation (SmF) was performed here. Three fungal colonies were isolated from biopharmaceutical oil waste collected from “Oushadhi” (The Pharmaceutical Corporation (IM) Kerala Ltd). The pure colonies were then used as inoculums for solid state and submerged fermentation of lipase. Strains were identified as Aspergillus sp., Trichoderma sp. and Penicillium sp. The oil waste itself was used as a substrate for SSF and it was supplemented with ((NH4)2SO4 5.0 g/l; Na2HPO4 6.0 g/l; KH2PO4 2.0 g/l; MgSO4.7H2O 3.0 g/l and CaCl2 3.0 g/l) at pH 6. The composition of production media (pH 6) used for SmF was glucose-10, peptone-20, NaCl-5 and yeast extract-5 (g/l).The comparison of enzyme yields showed that the yield in SSF was higher than that of SmF in most cases. Out of the three fungal strains, lipase produced by Aspergillus sp. showed the highest enzyme activity on assay by olive oil substrate emulsion method

Screening of Influenza a (H1N1) Neuraminidase Inhibitor for Kabasura Kudineer, Nilavembu Kudineer and the Novel Formulation JACOM

Due to erratic climate change, vector-borne diseases started flaring up from the second half of the last decade. Siddha medicine has been used as a public health tool to effectively manage chikungunya and dengue in the epidemics that happened in 2008 and 2016. Tamil Nadu government has made enormous efforts to control vector-borne diseases. Due to which morbidity and mortality due to vector borne diseases came down compared with other states. Two official Siddha formulations, namely Kabasura Kudineer Chooranam and Nilavembu Kudineer Chooranam and novel herbal formulation – JACOM, are used to combat vector-borne diseases. These decoctions lack an evidence base as a formulation. Screening has been done to check the efficacy of the formulation in inhibiting neuraminidase. Neuraminidase inhibition assay was performed to determine the activity of Siddha formulations. The Kabasura Kudineer Chooranam, Nilavembu Kudineer Chooranam and JACOM showed excellent inhibitory activity. The Kabasura Kudineer and Nilavembu Kudineer and JACOM aqueous extract showed maximum neuraminidase inhibition of 80.35%, 91.78% and 87.97%, respectively.</p