Screening, characterization, and kinetic studies of a serine alkaline protease from kitchen wastewater bacteria P2S1An and evaluation of its application in nutraceutical production.

Abstract

This present investigation aimed at characterizing the biochemical potential and kinetic study of the protease isolated from kitchen wastewater bacteria, P2S1An. The enzymatic activity was optimum when incubated for 96 h, at 30°C and pH9.0. The enzymatic activity of the purified protease (PrA) was 10.47-folds that of crude protease (S1). PrA was about 35 kDa in molecular weight. The broad pH and thermal stability, chelators, surfactants and solvent tolerance, and favorable thermodynamics suggested the potentiality of the extracted protease PrA. Thermal activity and stability were enhanced in presence of 1-mM Ca2+ ion at high temperatures. The protease was a serine one as its activity was completely diminished in presence of 1-mM PMSF. The Vmax , Km , and Kcat /Km suggested stability and catalytic efficiency of the protease. PrA hydrolyzes fish protein with 26.61 ±0.16% of peptide bond cleavage after 240 min, comparable to Alcalase 2.4L (27.13 ±0.31%). PRACTITIONER POINTS: A serine alkaline protease PrA was extracted from kitchen wastewater bacteria Bacillus tropicus Y14. Protease PrA showed significant activity and stability in a wide temperature and pH range. Protease showed great stability towards additives like metal ions, solvents, surfactants, polyols, and inhibitors. Kinetic study showed that the protease PrA had a prominent affinity and catalytic efficiency for the substrates. PrA hydrolysed fish proteins into short bioactive peptides which signify its potential in the formation of functional food ingredients.