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Characterization of a glutamate-cysteine ligase in Bombyx mori

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Abstract

Glutamate-cysteine ligase (GCL) is a crucial enzyme involved in the synthesis of glutathione (GSH). Despite various studies on glutathione transferase, and its essential role in detoxification and resistance to oxidative stress, GSH synthesis has not been described in Bombyx mori (silkworms) to date. Silkworms form part of the lepidopterans that are considered as a model of agricultural pests. This study aimed to understand the GSH synthesis by GCL in silkworms, which may help in developing insecticides to tackle agricultural pests. Based on the amino acid sequence and phylogenetic tree, the B. mori GCL belongs to group 2, and is designated bmGCL. Recombinant bmGCL was overexpressed and purified to ensure homogeneity. Biochemical studies revealed that bmGCL uses ATP and Mg2+ to ligate glutamate and cysteine. High expression levels of bmgcl mRNA and GSH were observed in the silkworm fat body after exposure to insecticides and UV-B irradiation. Moreover, we found an increase in bmgcl mRNA and GSH content during pupation in the silkworm fat body. In this study, we characterized the B. mori GCL and analyzed its biochemical properties. These observations indicate that bmGCL might play an important role in the resistance to oxidative stress in the silkworms.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

GSH:

Glutathione

GCL:

Glutamate-cysteine ligase

bmGSTO:

B. mori omega-class GST

GST:

GSH transferases

PCR:

Polymerase chain reaction

RT-PCR:

Reverse transcription-PCR

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Funding

The work was supported in part by the Research Grant for Young Investigators of Faculty of Agriculture, Kyushu University.

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Authors and Affiliations

  1. Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Faculty of Agriculture, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka, 819-0395, Japan

    Wazifa Afrin, Shigeki Furuya & Kohji Yamamoto

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  1. Wazifa Afrin
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  2. Shigeki Furuya
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  3. Kohji Yamamoto
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Contributions

WA and KY collected the data and performed data analysis; WA, SF and KY conceived and designed the study, and wrote the manuscript.

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Correspondence to Kohji Yamamoto.

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Afrin, W., Furuya, S. & Yamamoto, K. Characterization of a glutamate-cysteine ligase in Bombyx mori. Mol Biol Rep 50, 2623–2631 (2023). https://doi.org/10.1007/s11033-022-08191-6

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