Purification and Functional Characterization of a Soluble Trehalase in <i>Lissorhoptrus oryzophilus</i> (Coleoptera: Curculionidae)
Trehalase is the only enzyme known for the irreversible splitting of trehalose and plays a major role in insect growth and development. In this report, we describe a basic study of the trehalase gene fragment encoding a soluble trehalase from <i>Lissorhoptrus oryzophilus</i> (<i>Lo...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-09-01
|
| Series: | Insects |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2075-4450/13/10/867 |
| _version_ | 1797472597228650496 |
|---|---|
| author | Qingtai Wang Kui Fang Lizhong Qi Xiao Wang Yu Pan Yunshuo Li Jinghui Xi Juhong Zhang |
| author_facet | Qingtai Wang Kui Fang Lizhong Qi Xiao Wang Yu Pan Yunshuo Li Jinghui Xi Juhong Zhang |
| author_sort | Qingtai Wang |
| collection | DOAJ |
| description | Trehalase is the only enzyme known for the irreversible splitting of trehalose and plays a major role in insect growth and development. In this report, we describe a basic study of the trehalase gene fragment encoding a soluble trehalase from <i>Lissorhoptrus oryzophilus</i> (<i>LoTRE1</i>). Sequence alignment and phylogenetic analysis suggested that <i>LoTRE1</i> was similar to some known insect trehalases and belongs to the Coleoptera trehalase group. Additionally, <i>LoTRE1</i> was expressed mainly in the fat body. Purified protein was obtained using heterologous expression of <i>LoTRE1</i> in <i>Escherichia coli</i>, and the recombinant protein exhibited the ability to decompose trehalose. Enzyme–substrate docking indicated the potential involvement of other residues in the catalytic activity, in addition to Asp 333. Moreover, feeding of adults on <i>LoTRE1</i> dsRNA silenced the transcription of <i>LoTRE1</i> and thereby reduced the activity of trehalase and increased the trehalose content; it also led to a 12% death rate. This study reveals essential molecular features of trehalase and offers insights into the structural aspects of this enzyme, which might be related to its function. Taken together, the findings demonstrate that <i>LoTRE1</i> is indispensable for adults of this pest and provide a new target for the control of <i>L. oryzophilus</i>. |
| first_indexed | 2024-03-09T20:03:38Z |
| format | Article |
| id | doaj.art-31006e353bb5405ab14c2c29edccf33a |
| institution | Directory Open Access Journal |
| issn | 2075-4450 |
| language | English |
| last_indexed | 2024-03-09T20:03:38Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Insects |
| spelling | doaj.art-31006e353bb5405ab14c2c29edccf33a2023-11-24T00:37:25ZengMDPI AGInsects2075-44502022-09-01131086710.3390/insects13100867Purification and Functional Characterization of a Soluble Trehalase in <i>Lissorhoptrus oryzophilus</i> (Coleoptera: Curculionidae)Qingtai Wang0Kui Fang1Lizhong Qi2Xiao Wang3Yu Pan4Yunshuo Li5Jinghui Xi6Juhong Zhang7College of Plant Science, Jilin University, Changchun 130062, ChinaTechnical Center of Kunming Customs, Kunming 650228, ChinaCollege of Plant Science, Jilin University, Changchun 130062, ChinaCollege of Plant Science, Jilin University, Changchun 130062, ChinaCollege of Plant Science, Jilin University, Changchun 130062, ChinaCollege of Plant Science, Jilin University, Changchun 130062, ChinaCollege of Plant Science, Jilin University, Changchun 130062, ChinaCollege of Plant Science, Jilin University, Changchun 130062, ChinaTrehalase is the only enzyme known for the irreversible splitting of trehalose and plays a major role in insect growth and development. In this report, we describe a basic study of the trehalase gene fragment encoding a soluble trehalase from <i>Lissorhoptrus oryzophilus</i> (<i>LoTRE1</i>). Sequence alignment and phylogenetic analysis suggested that <i>LoTRE1</i> was similar to some known insect trehalases and belongs to the Coleoptera trehalase group. Additionally, <i>LoTRE1</i> was expressed mainly in the fat body. Purified protein was obtained using heterologous expression of <i>LoTRE1</i> in <i>Escherichia coli</i>, and the recombinant protein exhibited the ability to decompose trehalose. Enzyme–substrate docking indicated the potential involvement of other residues in the catalytic activity, in addition to Asp 333. Moreover, feeding of adults on <i>LoTRE1</i> dsRNA silenced the transcription of <i>LoTRE1</i> and thereby reduced the activity of trehalase and increased the trehalose content; it also led to a 12% death rate. This study reveals essential molecular features of trehalase and offers insights into the structural aspects of this enzyme, which might be related to its function. Taken together, the findings demonstrate that <i>LoTRE1</i> is indispensable for adults of this pest and provide a new target for the control of <i>L. oryzophilus</i>.https://www.mdpi.com/2075-4450/13/10/867<i>Lissorhoptrus oryzophilus</i>soluble trehalasemolecular dockingRNAiprokaryotic expressionhomology modeling |
| spellingShingle | Qingtai Wang Kui Fang Lizhong Qi Xiao Wang Yu Pan Yunshuo Li Jinghui Xi Juhong Zhang Purification and Functional Characterization of a Soluble Trehalase in <i>Lissorhoptrus oryzophilus</i> (Coleoptera: Curculionidae) Insects <i>Lissorhoptrus oryzophilus</i> soluble trehalase molecular docking RNAi prokaryotic expression homology modeling |
| title | Purification and Functional Characterization of a Soluble Trehalase in <i>Lissorhoptrus oryzophilus</i> (Coleoptera: Curculionidae) |
| title_full | Purification and Functional Characterization of a Soluble Trehalase in <i>Lissorhoptrus oryzophilus</i> (Coleoptera: Curculionidae) |
| title_fullStr | Purification and Functional Characterization of a Soluble Trehalase in <i>Lissorhoptrus oryzophilus</i> (Coleoptera: Curculionidae) |
| title_full_unstemmed | Purification and Functional Characterization of a Soluble Trehalase in <i>Lissorhoptrus oryzophilus</i> (Coleoptera: Curculionidae) |
| title_short | Purification and Functional Characterization of a Soluble Trehalase in <i>Lissorhoptrus oryzophilus</i> (Coleoptera: Curculionidae) |
| title_sort | purification and functional characterization of a soluble trehalase in i lissorhoptrus oryzophilus i coleoptera curculionidae |
| topic | <i>Lissorhoptrus oryzophilus</i> soluble trehalase molecular docking RNAi prokaryotic expression homology modeling |
| url | https://www.mdpi.com/2075-4450/13/10/867 |
| work_keys_str_mv | AT qingtaiwang purificationandfunctionalcharacterizationofasolubletrehalaseinilissorhoptrusoryzophilusicoleopteracurculionidae AT kuifang purificationandfunctionalcharacterizationofasolubletrehalaseinilissorhoptrusoryzophilusicoleopteracurculionidae AT lizhongqi purificationandfunctionalcharacterizationofasolubletrehalaseinilissorhoptrusoryzophilusicoleopteracurculionidae AT xiaowang purificationandfunctionalcharacterizationofasolubletrehalaseinilissorhoptrusoryzophilusicoleopteracurculionidae AT yupan purificationandfunctionalcharacterizationofasolubletrehalaseinilissorhoptrusoryzophilusicoleopteracurculionidae AT yunshuoli purificationandfunctionalcharacterizationofasolubletrehalaseinilissorhoptrusoryzophilusicoleopteracurculionidae AT jinghuixi purificationandfunctionalcharacterizationofasolubletrehalaseinilissorhoptrusoryzophilusicoleopteracurculionidae AT juhongzhang purificationandfunctionalcharacterizationofasolubletrehalaseinilissorhoptrusoryzophilusicoleopteracurculionidae |