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ID 55726
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Okazaki, Tomoaki Graduate School of Natural Science and TechnologyOkayama University
Ichinose, Junya Graduate School of Natural Science and TechnologyOkayama University
Takebe, So Graduate School of Biology-Oriented Science and TechnologyKindai University
Ide, Toru Graduate School of Natural Science and TechnologyOkayama University
Hayakawa, Tohru Graduate School of Natural Science and TechnologyOkayama University
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.
Keywords
Bacillus thuringiensis
Mosquitocidal Cry46Ab toxin
4AaCter-tag
Formation of protein inclusion
Escherichia coli
Note
This is an Accepted Manuscript of an article published by Springer Nature
Published Date
2018-02
Publication Title
Applied Entomology and Zoology
Volume
volume53
Issue
issue1
Publisher
Springer Japan
Start Page
67
End Page
73
ISSN
0003-6862
NCID
AA00543238
Content Type
Journal Article
language
英語
OAI-PMH Set
岡山大学
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isVersionOf https://doi.org/10.1007/s13355-017-0529-5