Biosynthesis of nano-inorganic materials by urease and phosphatase positive probiotic bacteria

Abstract

In recent years’ green synthesis has received more attention than chemical synthesis due to its eco-friendly and compatibility. In this study Bacillus subtilis and Bacillus coagulans, two potential probiotics that produce antimicrobial agents were used to synthesize nano-hydroxyapatite and nano-calcite. Grown Bacillus in medium culture containing insoluble calcium phosphate produced nano-hydroxyapatite and nano-crystalline calcite. The nano-hydroxyapatite was purified from nano-calcite by heat treatment and washing with a 200 nm filter. The structures, characteristics, and elemental analysis of nano-sized material were studied by Fourier Transform Infrared Spectroscopy, X-Ray Diffraction, Scanning Electron Microscope, Ultraviolet-visible spectroscopy, Energy dispersive X-Ray, and X-Ray Fluorescence. The results showed that hydroxyapatite is made only in a medium containing insoluble calcium phosphate sublimated with urea which is induced phosphatase and urease. Here, for the first time, the braided bacterial nano-hydroxyapatite similar to the bone structure was made in the medium which induced the production of urease and phosphatase (Maximum 99 U/L) enzymes, and the particle size was less than 100 nm. The ratio of calcium to phosphate in crude hydroxyapatite and calcite crystal particles made by Bacillus coagulans was 2.9, however, this ratio for pure hydroxyapatite was 1.7. Since the particles are made by antibacterial potential probiotics, the biological production of these particles makes them a suitable candidate to be used in toothpaste, bone regeneration, and sanitation products.