A ShK-like Domain from Steinernema carpocapsae with Bioinsecticidal Potential

Jorge Frias; Duarte Toubarro; Gro Elin Kjæreng Bjerga; Pål Puntervoll; João B. Vicente; Rui L. Reis; Nelson Simões

doi:10.3390/toxins14110754

A ShK-like Domain from Steinernema carpocapsae with Bioinsecticidal Potential

Jorge Frias, Duarte Toubarro, Gro Elin Kjæreng Bjerga, Pål Puntervoll, João B. Vicente, Rui L. Reis, Nelson Simões

Instituto de Tecnologia Química e Biológica António Xavier (ITQB)

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

Abstract

Entomopathogenic nematodes are used as biological control agents against a broad range of insect pests. We ascribed the pathogenicity of these organisms to the excretory/secretory products (ESP) released by the infective nematode. Our group characterized different virulence factors produced by Steinernema carpocapsae that underlie its success as an insect pathogen. A novel ShK-like peptide (ScK1) from this nematode that presents high sequence similarity with the ShK peptide from a sea anemone was successfully produced recombinantly in Escherichia coli. The secondary structure of ScK1 appeared redox-sensitive, exhibiting a far-UV circular dichroism spectrum consistent with an alpha-helical secondary structure. Thermal denaturation of the ScK1 allowed estimating the melting temperature to 59.2 ± 0.1 °C. The results from toxicity assays using Drosophila melanogaster as a model show that injection of this peptide can kill insects in a dose-dependent manner with an LD₅₀ of 16.9 µM per adult within 24 h. Oral administration of the fusion protein significantly reduced the locomotor activity of insects after 48 h (p < 0.05, Tukey’s test). These data show that this nematode expresses insecticidal peptides with potential as next-generation insecticides.

Original language	English
Article number	754
Journal	Toxins
Volume	14
Issue number	11
DOIs	https://doi.org/10.3390/toxins14110754
Publication status	Published - Nov 2022

Keywords

disulfide-rich peptide
entomopathogenic nematodes
excretory/secretory products
fruit fly
insecticidal toxins
locomotor activity
ShK
toxicity assay

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10.3390/toxins14110754

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@article{ecd6cf0e0587437c8b70aec99b663b42,

title = "A ShK-like Domain from Steinernema carpocapsae with Bioinsecticidal Potential",

abstract = "Entomopathogenic nematodes are used as biological control agents against a broad range of insect pests. We ascribed the pathogenicity of these organisms to the excretory/secretory products (ESP) released by the infective nematode. Our group characterized different virulence factors produced by Steinernema carpocapsae that underlie its success as an insect pathogen. A novel ShK-like peptide (ScK1) from this nematode that presents high sequence similarity with the ShK peptide from a sea anemone was successfully produced recombinantly in Escherichia coli. The secondary structure of ScK1 appeared redox-sensitive, exhibiting a far-UV circular dichroism spectrum consistent with an alpha-helical secondary structure. Thermal denaturation of the ScK1 allowed estimating the melting temperature to 59.2 ± 0.1 °C. The results from toxicity assays using Drosophila melanogaster as a model show that injection of this peptide can kill insects in a dose-dependent manner with an LD50 of 16.9 µM per adult within 24 h. Oral administration of the fusion protein significantly reduced the locomotor activity of insects after 48 h (p < 0.05, Tukey{\textquoteright}s test). These data show that this nematode expresses insecticidal peptides with potential as next-generation insecticides.",

keywords = "disulfide-rich peptide, entomopathogenic nematodes, excretory/secretory products, fruit fly, insecticidal toxins, locomotor activity, ShK, toxicity assay",

author = "Jorge Frias and Duarte Toubarro and Bjerga, {Gro Elin Kj{\ae}reng} and P{\aa}l Puntervoll and Vicente, {Jo{\~a}o B.} and Reis, {Rui L.} and Nelson Sim{\~o}es",

note = "Funding Information: J.F. received a doctoral grant from the FCT (SFRH/BD/131698/2017). This work was funded by Fundo Europeu de Desenvolvimento Regional (FEDER) under project NanoNema (ACORES-01-0145-FEDER000113), approved by the Autoridade de Gest{\~a}o do Programa Operacional A{\c C}ORES 2020. J.B.V. acknowledges support by FCT-Funda{\c c}{\~a}o para a Ci{\^e}ncia e a Tecnologia, Portuguese Institute (P.I), through iNOVA4Health (UIDB/04462/2020, UIDP/04462/2020) and LS4FUTURE Associated Laboratory (LA/P/0087/2020). G.B. and P.P. acknowledge funding from NORCE Norwegian Research Centre to host JF as well as funding from the Research Council of Norway (Grant ID: 221568) for expansion of the expression vector suite. Publisher Copyright: {\textcopyright} 2022 by the authors.",

year = "2022",

month = nov,

doi = "10.3390/toxins14110754",

language = "English",

volume = "14",

journal = "Toxins",

issn = "2072-6651",

publisher = "MDPI - Multidisciplinary Digital Publishing Institute",

number = "11",

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T1 - A ShK-like Domain from Steinernema carpocapsae with Bioinsecticidal Potential

AU - Frias, Jorge

AU - Toubarro, Duarte

AU - Bjerga, Gro Elin Kjæreng

AU - Puntervoll, Pål

AU - Vicente, João B.

AU - Reis, Rui L.

AU - Simões, Nelson

N1 - Funding Information: J.F. received a doctoral grant from the FCT (SFRH/BD/131698/2017). This work was funded by Fundo Europeu de Desenvolvimento Regional (FEDER) under project NanoNema (ACORES-01-0145-FEDER000113), approved by the Autoridade de Gestão do Programa Operacional AÇORES 2020. J.B.V. acknowledges support by FCT-Fundação para a Ciência e a Tecnologia, Portuguese Institute (P.I), through iNOVA4Health (UIDB/04462/2020, UIDP/04462/2020) and LS4FUTURE Associated Laboratory (LA/P/0087/2020). G.B. and P.P. acknowledge funding from NORCE Norwegian Research Centre to host JF as well as funding from the Research Council of Norway (Grant ID: 221568) for expansion of the expression vector suite. Publisher Copyright: © 2022 by the authors.

PY - 2022/11

Y1 - 2022/11

N2 - Entomopathogenic nematodes are used as biological control agents against a broad range of insect pests. We ascribed the pathogenicity of these organisms to the excretory/secretory products (ESP) released by the infective nematode. Our group characterized different virulence factors produced by Steinernema carpocapsae that underlie its success as an insect pathogen. A novel ShK-like peptide (ScK1) from this nematode that presents high sequence similarity with the ShK peptide from a sea anemone was successfully produced recombinantly in Escherichia coli. The secondary structure of ScK1 appeared redox-sensitive, exhibiting a far-UV circular dichroism spectrum consistent with an alpha-helical secondary structure. Thermal denaturation of the ScK1 allowed estimating the melting temperature to 59.2 ± 0.1 °C. The results from toxicity assays using Drosophila melanogaster as a model show that injection of this peptide can kill insects in a dose-dependent manner with an LD50 of 16.9 µM per adult within 24 h. Oral administration of the fusion protein significantly reduced the locomotor activity of insects after 48 h (p < 0.05, Tukey’s test). These data show that this nematode expresses insecticidal peptides with potential as next-generation insecticides.

AB - Entomopathogenic nematodes are used as biological control agents against a broad range of insect pests. We ascribed the pathogenicity of these organisms to the excretory/secretory products (ESP) released by the infective nematode. Our group characterized different virulence factors produced by Steinernema carpocapsae that underlie its success as an insect pathogen. A novel ShK-like peptide (ScK1) from this nematode that presents high sequence similarity with the ShK peptide from a sea anemone was successfully produced recombinantly in Escherichia coli. The secondary structure of ScK1 appeared redox-sensitive, exhibiting a far-UV circular dichroism spectrum consistent with an alpha-helical secondary structure. Thermal denaturation of the ScK1 allowed estimating the melting temperature to 59.2 ± 0.1 °C. The results from toxicity assays using Drosophila melanogaster as a model show that injection of this peptide can kill insects in a dose-dependent manner with an LD50 of 16.9 µM per adult within 24 h. Oral administration of the fusion protein significantly reduced the locomotor activity of insects after 48 h (p < 0.05, Tukey’s test). These data show that this nematode expresses insecticidal peptides with potential as next-generation insecticides.

KW - disulfide-rich peptide

KW - entomopathogenic nematodes

KW - excretory/secretory products

KW - fruit fly

KW - insecticidal toxins

KW - locomotor activity

KW - ShK

KW - toxicity assay

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U2 - 10.3390/toxins14110754

DO - 10.3390/toxins14110754

M3 - Article

C2 - 36356004

AN - SCOPUS:85141810971

SN - 2072-6651

VL - 14

JO - Toxins

JF - Toxins

IS - 11

M1 - 754

ER -

A ShK-like Domain from Steinernema carpocapsae with Bioinsecticidal Potential

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Keywords

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