Functional analysis of the gene SCO1782 encoding Streptomyces hemolysin (S-hemolysin) in Streptomyces coelicolor M145

Toxicon. 2013 Sep:71:159-65. doi: 10.1016/j.toxicon.2013.05.023. Epub 2013 Jun 4.

Abstract

In the process of evaluating the growth of Streptomyces coelicolor on rich media such as blood agar, we found that S. coelicolor a non-pathogenic, well-known antibiotic producer had the ability to grow and produce a prominent hemolytic zone. By comparing the growth with an agarase gene mutant of S. coelicolor, a similar prominent hemolytic zone was found to develop due to the organism's hemolytic activity. After the confirmation of hemolytic activity from S. coelicolor, the genome was searched for hemolysin-coding genes; consequently, SCO1782, SCO2534, and SCO3882 were identified, whose products were annotated as a putative, membrane, and hypothetical proteins, respectively. Functional characterization of all the recombinant proteins expressed in Escherichia coli BL21(DE3) revealed that only SCO1782 exhibited hemolytic activity. This S. coelicolor protein, designated as S-hemolysin, showed sequence similarity toward hemolysins from Brachyspira hyodysenteriae (35%) and Mycobacterium tuberculosis (62%). Recombinant hemolysin exhibited activity against sheep blood erythrocytes and cytolytic activity against human fibroblast cells. Deletion of SCO1782 resulted in complete loss of hemolysin activity in S. coelicolor.

Keywords: Cytolytic activity; Hemolytic activity; S-hemolysin; Streptomyces coelicolor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Bacterial Agents / metabolism
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics*
  • Cloning, Molecular
  • Culture Media
  • Erythrocytes / drug effects
  • Erythrocytes / metabolism
  • Escherichia coli / genetics
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Gene Deletion
  • Genes, Bacterial*
  • Hemolysin Proteins / chemistry
  • Hemolysin Proteins / genetics*
  • Hemolytic Agents / pharmacology
  • Humans
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Sheep
  • Streptomyces coelicolor / chemistry
  • Streptomyces coelicolor / genetics*

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Culture Media
  • Hemolysin Proteins
  • Hemolytic Agents
  • Recombinant Proteins