Agricultural waste materials enhance protease production by Bacillus subtilis B22 in submerged fermentation under blue light-emitting diodes

Abstract

Bacillus bacteria have major utility in large-scale production of industrial enzymes, among which proteases have particular importance. B. subtilis B22, an aerobic and chemotrophic strain, was isolated from kimchi and identified by 16S rRNA gene sequencing. Extracellular protease production was determined in basic medium, with 1% (w/v) casein as substrate, by submerged fermentation at 37 °C under blue, green, red and white light-emitting diodes (LEDs), white fluorescent light and darkness. Fermentation under blue LEDs maximized protease production (110.79 ± 1.8 U/mL at 24 h). Various agricultural waste products enhanced production and groundnut oil cake yielded the most protease (334 ± 1.8 U/mL at 72 h). Activity and stability of the purified protease were optimum at pH 7-10 and 20-60 °C. Activity increased in the presence of Ca2+, Mg2+ and Mn2+, while Fe2+, Zn2+, Co2+ and Cu2+ moderated activity, and Ni2+ and Hg2+ inhibited activity. Activity was high (98%) in the presence of ethylenediaminetetraacetic acid (EDTA) but inhibited by phenylmethanesulfonyl fluoride (PMSF). The protease was unaffected by nonionic surfactants, tolerated an anionic surfactant and oxidizing agents, and was compatible with multiple organic solvents. These properties suggest utility of protease produced by B. subtilis B22 under blue LEDs for industrial applications.

Keywords: Agricultural waste products; B. subtilis; Blue LEDs; Protease; Submerged fermentation.