Optimization, partial purification, and characterization of a novel high molecular weight alkaline protease produced by Halobacillus sp. HAL1 using fish wastes as a substrate.

BACKGROUND: Hydrolytic enzymes from halophilic microorganisms have a wide range of industrial applications. Herein, we report the isolation of Halobacillus sp. HAL1, a moderately halophilic bacterium that produces a novel high molecular weight extracellular alkaline protease when grown in fish processing wastes as a substrate. RESULTS: Results showed that the isolated strain belonged to the genus Halobacillus, and it was designated as Halobacillus sp. HAL1 with the GenBank accession number OK001470. The strain secreted an extracellular alkaline protease, and the highest yield was obtained when it was grown in a medium with fish wastes substrate as the sole nutritional source (10 g/L) and incubated at 25 °C under shaking conditions. The enzyme was partially purified by Sephadex G-100 column chromatography. Zymographic analysis showed two casein degrading bands of about 190 and 250 KDa. The optimum enzyme activity was at a temperature of 50 °C at pH 8. The proteolytic activity was enhanced in the presence of metal ions (Ca2+, Mg2+, and Mn2+), surfactants (Tween 80, SDS, and Triton-X100), H2O2, and EDTA. CONCLUSION: Our study indicates that Haobacillus sp. HAL1 is a moderately halophilic strain and secrets a novel high molecular wight alkaline protease that is suitable for detergent formulation.

Green Synthesis of Zinc Oxide Nanoparticles (ZnO-NPs) by Pseudomonas aeruginosa and Their Activity against Pathogenic Microbes and Common House Mosquito, Culex pipiens.

The synthesis of nanoparticles by green approaches is gaining unique importance due to its low cost, biocompatibility, high productivity, and purity, and being environmentally friendly. Herein, biomass filtrate of Pseudomonas aeruginosa isolated from mangrove rhizosphere sediment was used for the biosynthesis of zinc oxide nanoparticles (ZnO-NPs). The bacterial isolate was identified based on morphological, physiological, and 16S rRNA. The bio-fabricated ZnO-NPs were characterized using color change, UV-visible spectroscopy, FT-IR, TEM, and XRD analyses. In the current study, spherical and crystalline nature ZnO-NPs were successfully formed at a maximum SPR (surface plasmon resonance) of 380 nm. The bioactivities of fabricated ZnO-NPs including antibacterial, anti-candida, and larvicidal efficacy were investigated. Data analysis showed that these bioactivities were concentration-dependent. The green-synthesized ZnO-NPs exhibited high efficacy against pathogenic Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis), Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa), and unicellular fungi (Candida albicans) with inhibition zones of (12.33 ± 0.9 and 29.3 ± 0.3 mm), (19.3 ± 0.3 and 11.7 ± 0.3 mm), and (22.3 ± 0.3 mm), respectively, at 200 ppm. The MIC value was detected as 50 ppm for E. coli, B. subtilis, and C. albicans, and 200 ppm for S. aureus and P. aeruginosa with zones of inhibition ranging between 11.7 ± 0.3-14.6 ± 0.6 mm. Moreover, the biosynthesized ZnO-NPs showed high mortality for Culex pipiens with percentages of 100 ± 0.0% at 200 ppm after 24 h as compared with zinc acetate (44.3 ± 3.3%) at the same concentration and the same time.