Potency of the mosquitocidal Cry46Ab toxin produced using a 4AaCter-tag, which facilitates formation of protein inclusion bodies in Escherichia coli

Tomoaki Okazaki, Junya Ichinose, So Takebe, Toru Ide, Tohru Hayakawa

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

A Cry46Ab toxin derived from Bacillus thuringiensis strain TK-E6 shows mosquitocidal activity against Culex pipiens pallens Coquillett (Diptera: Culicidae) larvae as well as preferential cytotoxicity against human cancer cells. In B. thuringiensis cells, Cry46Ab is produced and accumulates as a protein crystal that is processed into the active 29-kDa toxin upon solubilization in the alkaline environment of the insect midgut. The Cry46Ab protoxin is 30 kDa, and is therefore thought to require an accessory protein such as P20 and/or ORF2 for efficient crystal formation. In the present study, the potency of the 4AaCter-tag was investigated for the production of alkali-soluble inclusion bodies of recombinant Cry46Ab in Escherichia coli. The 4AaCter-tag is a polypeptide derived from the C-terminal region of the B. thuringiensis Cry4Aa toxin and facilitates the formation of alkali-soluble protein inclusion bodies in E. coli. Fusion with the 4AaCter-tag enhanced both Cry46Ab production and the formation of Cry46Ab inclusion bodies. In addition, upon optimization of protein expression procedures, the Cry46Ab–4AaCter inclusion bodies showed mosquitocidal activity and stability in aqueous environments comparable to Cry46Ab without the 4AaCter-tag. Our study suggests that use of the 4AaCter-tag is a straightforward approach for preparing formulations of smaller-sized Cry toxins such as Cry46Ab in E. coli.

Original languageEnglish
Pages (from-to)67-73
Number of pages7
JournalApplied Entomology and Zoology
Volume53
Issue number1
DOIs
Publication statusPublished - Feb 1 2018

Fingerprint

inclusion bodies
toxins
Bacillus thuringiensis
Escherichia coli
alkalis
proteins
Culex pipiens pallens
crystal proteins
solubilization
midgut
crystals
cytotoxicity
polypeptides
Culicidae
protein synthesis
insects
larvae
water
cells

Keywords

  • 4AaCter-tag
  • Bacillus thuringiensis
  • Escherichia coli
  • Formation of protein inclusion
  • Mosquitocidal Cry46Ab toxin

ASJC Scopus subject areas

  • Insect Science

Cite this

Potency of the mosquitocidal Cry46Ab toxin produced using a 4AaCter-tag, which facilitates formation of protein inclusion bodies in Escherichia coli. / Okazaki, Tomoaki; Ichinose, Junya; Takebe, So; Ide, Toru; Hayakawa, Tohru.

In: Applied Entomology and Zoology, Vol. 53, No. 1, 01.02.2018, p. 67-73.

Research output: Contribution to journalArticle

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