Hyaluronidase from Paenibacillus aquistagni SH-7-A: Identification and characterization

Abstract

The research identified a hyaluronidase produced by Paenibacillus aquistagni SH-7-A, the highest hyaluronidase activity with 830 U/mL obtained under 24 h fermentation. Based on genomic analysis, a gene hysa1 encoding hyaluronidase hysA1 was mined. Hyaluronidase H1 was purified by ammonium sulfate precipitation, anion exchange chromatography, and gel permeation chromatography. The molecular weight of purified H1 was determined to be about 110 kDa by SDS-PAGE, and the sequence coverage of the protein search with hysA1 resulted in 77% by HPLC-MS/MS. Therefore, it was proven that hyaluronidase H1 was hysA1. The optimum temperature and pH for H1 activity were 40°C and pH 6, and it was stable at 30–40°C and pH 5–7, respectively. Ca2+, Mg2+, and Ni2+ enhanced the enzymatic activity, and it was strongly inhibited by Zn2+, Cu2+, EDTA, tween 80, Triton X-100, CTAB, DTT, and SDS. Using HA as a substrate, the Km and Vmax of H1 were found to be 0.1124 mg/mL and 0.0184 mol/min/mL, respectively. This is the first hyaluronidase production, purification, and characterization report from P.aquistagni. These characteristics showed that the hyaluronidase H1 would provide potential possibilities for commercial applications.