Cloning and Biochemical Characterization of a Hyaluronate Lyase from Bacillus sp. CQMU-D

J Microbiol Biotechnol. 2023 Feb 28;33(2):235-241. doi: 10.4014/jmb.2209.09036. Epub 2022 Dec 7.

Abstract

Hyaluronidase (HAase) can enhance drug diffusion and dissipate edema by degrading hyaluronic acid (HA) in the extracellular matrix into unsaturated HA oligosaccharides in mammalian tissues. Microorganisms are recognized as valuable sources of HAase. In this study, a new hyaluronate lyase (HAaseD) from Bacillus sp. CQMU-D was expressed in Escherichia coli BL21, purified, and characterized. The results showed that HAaseD belonged to the polysaccharide lyase (PL) 8 family and had a molecular weight of 123 kDa. HAaseD could degrade chondroitin sulfate (CS) -A, CS-B, CS-C, and HA, with the highest activity toward HA. The optimum temperature and pH value of HAaseD were 40°C and 7.0, respectively. In addition, HAaseD retained stability in an alkaline environment and displayed higher activity with appropriate concentrations of metal ions. Moreover, HAaseD was an endolytic hyaluronate lyase that could degrade HA to produce unsaturated HA oligosaccharides. Together, our findings indicate that HAaseD from Bacillus sp. CQMU-D is a new hyaluronate lyase and with excellent potential for application in industrial production.

Keywords: Bacillus; Hyaluronate lyase; characterization; hyaluronic acid.

MeSH terms

  • Animals
  • Bacillus* / genetics
  • Cloning, Molecular
  • Hyaluronic Acid / metabolism
  • Hyaluronoglucosaminidase / metabolism
  • Hydrogen-Ion Concentration
  • Mammals / metabolism
  • Oligosaccharides / metabolism
  • Polysaccharide-Lyases / metabolism
  • Substrate Specificity

Substances

  • hyaluronate lyase
  • Polysaccharide-Lyases
  • Hyaluronoglucosaminidase
  • Hyaluronic Acid
  • Oligosaccharides