ABSTRACT
The aim of this study was to use agro-industrial residues for the production of a halotolerant keratinolytic- protease by Actinobacterium sp. in solid-state fermentation. Among various agro-industrial residues that were evaluated, apple pomace supported maximum protease production (8400 U/g material). The optimum conditions required for enzyme production were a fermentation period of 72 h, 10% (w/v) NaCl, pH 7.0, 120% (v/w) moisture and 10% (v/w) inoculum. The enzyme exhibited activity to a range of pH (7.0-9.0) and temperature (30-45°C), with optima at 8.0 and 40 °C, respectively. Most of the divalent ions tested stimulated the protease activity and Ca2+ ion was required for its activity and stability. The enzyme was widely active at the range of NaCl concentration (5%-15%, w/v) and effectively degraded chicken feather. This protease could be useful in fish sauce fermentation and also in feed industry.
ABSTRACT
<p><b>OBJECTIVE</b>The aim of the present study was to isolate the anti-MRSA (Methicillin Resistant Staphylococcus aureus) molecule from the Mangrove symbiont Streptomyces and its biomedical studies in Zebrafish embryos.</p><p><b>METHODS</b>MRSA was isolated from the pus samples of Colachal hospitals and confirmed by amplification of mecA gene. Anti-MRSA molecule producing strain was identified by 16s rRNA gene sequencing. Anti-MRSA compound production was optimized by Solid State Fermentation (SSF) and the purification of the active molecule was carried out by TLC and RP-HPLC. The inhibitory concentration and LC50 were calculated using Statistical software SPSS. The Biomedical studies including the cardiac assay and organ toxicity assessment were carried out in Zebrafish.</p><p><b>RESULTS</b>The bioactive anti-MRSA small molecule A2 was purified by TLC with Rf value of 0.37 with 1.389 retention time at RP-HPLC. The Inhibitory Concentration of the purified molecule A2 was 30 µg/mL but, the inhibitory concentration of the MRSA in the infected embryo was 32-34 µg/mL for TLC purified molecule A2 with LC50 mean value was 61.504 µg/mL. Zebrafish toxicity was assessed in 48-60 µg/mL by observing the physiological deformities and the heart beat rates (HBR) of embryos for anti MRSA molecule showed the mean of 41.33-41.67 HBR/15 seconds for 40 µg/mL and control was 42.33-42.67 for 15 seconds which significantly showed that the anti-MRSA molecule A2 did not affected the HBR.</p><p><b>CONCLUSIONS</b>Anti-MRSA molecule from Streptomyces sp PVRK-1 was isolated and biomedical studies in Zebrafish model assessed that the molecule was non toxic at the minimal inhibitory concentration of MRSA.</p>