Biotechnologically Relevant Enzymes and Proteins

Applied Microbiology and Biotechnology

, Volume 72, Issue 4, pp 713-719

First online:

Discovery and characterization of Sip1A: a novel secreted protein from Bacillus thuringiensis with activity against coleopteran larvae

  • William P. DonovanAffiliated withMonsanto Company Email author 
  • , James T. EnglemanAffiliated withEcogen Inc.
  • , Judith C. DonovanAffiliated withEcogen Inc.
  • , James A. BaumAffiliated withMonsanto Company
  • , Greg J. BunkersAffiliated withMonsanto Company
  • , David J. ChiAffiliated withMonsanto Company
  • , William P. ClintonAffiliated withMonsanto Company
  • , Leigh EnglishAffiliated withMonsanto Company
  • , Gregory R. HeckAffiliated withMonsanto Company
    • , Oliver M. IlaganAffiliated withMonsanto Company
    • , Karina C. Krasomil-OsterfeldAffiliated withMonsanto Company
    • , John W. PitkinAffiliated withMonsanto Company
    • , James K. RobertsAffiliated withMonsanto Company
    • , Matthew R. WaltersAffiliated withMonsanto Company

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Bioassay screening of Bacillus thuringiensis culture supernatants identified strain EG2158 as having larvicidal activity against Colorado potato beetle (Leptinotarsa decemlineata) larvae. Ion-exchange fractionation of the EG2158 culture supernatant resulted in the identification of a protein designated Sip1A (secreted insecticidal protein) of approximately 38 kDa having activity against Colorado potato beetle (CPB). An oligonucleotide probe based on the N-terminal sequence of the purified Sip1A protein was used to isolate the sip1A gene. The sequence of the Sip1A protein, as deduced from the sequence of the cloned sip1A gene, contained 367 residues (41,492 Da). Recombinant B. thuringiensis and Escherichia coli harboring cloned sip1A produced Sip1A protein which had insecticidal activity against larvae of CPB, southern corn rootworm (Diabrotica undecimpunctata howardi), and western corn rootworm (Diabrotica virgifera virgifera).