Interactions between Hyaluronic Acid, Lysozyme, Peroxidase, and Glucose Oxidase in Enzymatic Activities at Low pH

Purpose: Many substances in saliva or oral health care products interact with each other. The aim of this study was to investigate interactions between hyaluronic acid (HA), lysozyme, peroxidase, and glucose oxidase (GO) in enzymatic activities at low pH levels. Methods: HA (0.5 mg/mL), hen egg-white lysozyme (HEWL, $30{\mu}g/mL$), bovine lactoperoxidase (bLPO, $25{\mu}g/mL$), and GO ($50{\mu}g/mL$) were used. The influences of HA, bLPO, and GO on HEWL activity were determined by measuring the turbidity of a Micrococcus lysodeikticus suspension. The influences of HA and HEWL on bLPO activity were determined by the NbsSCN assay, measuring the rate of oxidation of 5-thio-2-nitrobenzoic acid (Nbs) to 5,5'-dithiobis(2-nitrobenzoic acid) $(Nbs)_2$. The influences of HA and HEWL on GO activity were determined by measuring oxidized o-dianisidine production. All experiments were performed at pH 4, 5, and 6. Results: HA and GO did not affect the enzymatic activity of HEWL at pH 4, 5, and 6. bLPO enhanced the enzymatic activity of HEWL at pH 5 (p<0.05) and pH 6 (p<0.05) significantly. The enzymatic activity of bLPO was not affected by HA and HEWL at pH 4, 5, and 6. HA and HEWL did not affect the enzymatic activity of the GO at pH 4, 5, and 6. Conclusions: Peroxidase enhances lysozyme activity at low pH, otherwise there were no significant interactions in enzymatic activities between HA, lysozyme, peroxidase, and GO at low pH levels.