Molecular and enzymatic characterization of betaC-S lyase in Streptococcus constellatus

Oral Microbiol Immunol. 2008 Jun;23(3):245-53. doi: 10.1111/j.1399-302X.2007.00419.x.

Abstract

Background/aims: Streptococcus anginosus and Streptococcus constellatus are frequently isolated from dental abscesses and other suppurative lesions. We previously reported that betaC-S lyase from a strain of S. anginosus produced significantly more hydrogen sulfide than betaC-S lyases from other streptococci. The purpose of this study was to establish the molecular and enzymatic features of the betaC-S lyase in S. constellatus and to elucidate whether this unique capacity is common to many strains of S. constellatus and S. anginosus.

Methods: The capacity of crude extract to produce hydrogen sulfide was evaluated among 16 strains of S. constellatus, S. anginosus, and Streptococcus gordonii. The lcd gene encoding betaC-S lyase was cloned from the genomic DNA of each strain to compare the deduced amino acid sequences. The recombinant betaC-S lyases of three representative strains were purified and characterized.

Results: Incubation of crude extracts from all strains of S. constellatus and S. anginosus with l-cysteine resulted in the production of a large amount of hydrogen sulfide. The primary sequence of betaC-S lyase was very similar among strains of S. constellatus and S. anginosus. The kinetic properties of the betaC-S lyases purified from S. constellatus resembled those for betaC-S lyases purified from S. anginosus. In contrast, the betaC-S lyases of S. constellatus and S. gordonii differed in terms of their hydrogen sulfide production, with the former producing much more.

Conclusion: A high level of hydrogen sulfide production, which appears to be a common feature in both S. constellatus and S. anginosus, may be associated with their abscess formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon-Sulfur Lyases / analysis*
  • Carbon-Sulfur Lyases / antagonists & inhibitors
  • Carbon-Sulfur Lyases / genetics
  • Coloring Agents
  • Consensus Sequence / genetics
  • Cystathionine / analysis
  • Cysteine / metabolism
  • DNA, Bacterial / analysis
  • Enzyme Inhibitors / pharmacology
  • Genome, Bacterial / genetics
  • Humans
  • Hydrogen Sulfide / analysis
  • Methylene Blue
  • Molecular Biology
  • Pyruvates / analysis
  • RNA, Ribosomal, 16S / analysis
  • Sequence Analysis, Protein
  • Sequence Homology, Amino Acid
  • Streptococcus anginosus / enzymology
  • Streptococcus constellatus / enzymology*
  • Streptococcus gordonii / enzymology

Substances

  • Coloring Agents
  • DNA, Bacterial
  • Enzyme Inhibitors
  • Pyruvates
  • RNA, Ribosomal, 16S
  • Cystathionine
  • Carbon-Sulfur Lyases
  • Cysteine
  • Methylene Blue
  • Hydrogen Sulfide