Increased toxicity of modified mosquitocidal binary toxins of Bacillus sphaericus expressed in Escherichia coli

Appl Microbiol Biotechnol. 1998 Feb;49(2):164-7. doi: 10.1007/s002530051153.

Abstract

The binary mosquitocidal genes of 51-kDa and 42-kDa proteins isolated from Bacillus sphaericus 1593 have been expressed at moderate levels in Escherichia coli employing the pQE expression system. The expressed proteins are readily visible in Coomassie blue-stained protein gels. The recombinant E. coli cells expressing toxic proteins were toxic towards Culex larvae. During the assembly of crystals in B. sphaericus, the 42-kDa toxin is first cleaved at the N-terminal end by a specific B. sphaericus protease. To express the toxins in E. coli the B. sphaericus specific protease-recognition site was deleted at the N-terminal end of the 42-kDa toxin, thereby mimicking the structure of the toxin as present in the crystal. This modification resulted in a twofold increase in the toxicity of the E. coli cells expressing the modified 42-kDa toxin as a constituent of the binary toxin. Our results demonstrate the utility of this modification for heterologous expression of the binary toxin genes from B. sphaericus.

MeSH terms

  • Animals
  • Bacillus / genetics*
  • Bacterial Toxins / biosynthesis*
  • Bacterial Toxins / genetics
  • Bacterial Toxins / toxicity*
  • Culex*
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics*
  • Gene Expression
  • Larva
  • Plasmids
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / toxicity

Substances

  • Bacterial Toxins
  • Recombinant Proteins
  • binB protein, Bacillus sphaericus