Mining and Expression of a Metagenome-Derived Keratinase Responsible for Biosynthesis of Silver Nanoparticles

Abstract

A keratinase gene <i>kerBv</i> was mined from soil metagenomes. The open reading frame consisted of 1149 bp and potentially encoded a protein of 382 amino acid residues. It shared the same active site with several reported typical keratinases via analysis of the amino acid sequence. The keratinase was successfully expressed in <i>B. subtilis</i> WB600 with pMA5 expression vector. The maximum activity of 164.8 U/mL in the fermentation supernatant was observed after incubating for 30 h in Terrifc Broth (TB) medium. The keratinase exhibited outstanding resistance to metal ions and was surfactant-stable. Additionally, the enzyme displayed broad substrate specificity especially toward insoluble substrate feather meal because of its disulfide bond-reducing activity. Furthermore, the reducing power of the recombinant keratinase was investigated. It showed that the protein exhibited a relatively high reducing power, which was subsequently used in the biosynthesis of silver nanoparticles (AgNPs). The biosynthesized AgNPs were characterized by ultraviolet-visible (UV-vis) spectroscopy, dynamic light scattering (DLS), transmission electron microscope (TEM), as well as Fourier transform infrared spectroscopy (FTIR) and displayed obvious antibacterial activities toward <i>Escherichia coli</i>.