start-ver=1.4
cd-journal=joma
no-vol=53
cd-vols=
no-issue=1
article-no=
start-page=67
end-page=73
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2018
dt-pub=201802
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Potency of the mosquitocidal Cry46Ab toxin produced using a 4AaCter-tag, which facilitates formation of protein inclusion bodies in Escherichia coli
en-subtitle=
kn-subtitle=
en-abstract=
kn-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.
en-copyright=
kn-copyright=

en-aut-name=OkazakiTomoaki
en-aut-sei=Okazaki
en-aut-mei=Tomoaki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=

en-aut-name=IchinoseJunya
en-aut-sei=Ichinose
en-aut-mei=Junya
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=

en-aut-name=TakebeSo
en-aut-sei=Takebe
en-aut-mei=So
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=

en-aut-name=IdeToru
en-aut-sei=Ide
en-aut-mei=Toru
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=

en-aut-name=HayakawaTohru
en-aut-sei=Hayakawa
en-aut-mei=Tohru
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=

affil-num=1
en-affil=Graduate School of Natural Science and TechnologyOkayama University
kn-affil=

affil-num=2
en-affil=Graduate School of Natural Science and TechnologyOkayama University
kn-affil=

affil-num=3
en-affil=Graduate School of Biology-Oriented Science and TechnologyKindai University
kn-affil=

affil-num=4
en-affil=Graduate School of Natural Science and TechnologyOkayama University
kn-affil=

affil-num=5
en-affil=Graduate School of Natural Science and TechnologyOkayama University
kn-affil=
en-keyword=Bacillus thuringiensis
kn-keyword=Bacillus thuringiensis
en-keyword=Mosquitocidal Cry46Ab toxin
kn-keyword=Mosquitocidal Cry46Ab toxin
en-keyword=4AaCter-tag
kn-keyword=4AaCter-tag
en-keyword=Formation of protein inclusion
kn-keyword=Formation of protein inclusion
en-keyword=Escherichia coli 
kn-keyword=Escherichia coli 
END