Expression of kiwifruit-derived actinidin in Nicotiana benthamiana leaves
Kiwifruit (Actinidia deliciosa)-derived actinidin, a cysteine protease, is renowned for its meat-tenderizing and milk-clotting activities. Despite its potential in various biotechnological applications, an efficient expression platform for actinidin production has not yet been developed. Instead, ac...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2024.1532170/full |
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| author | Ji Hyun Kang Jae-Ho Lee Dong Wook Lee Dong Wook Lee Dong Wook Lee |
| author_facet | Ji Hyun Kang Jae-Ho Lee Dong Wook Lee Dong Wook Lee Dong Wook Lee |
| author_sort | Ji Hyun Kang |
| collection | DOAJ |
| description | Kiwifruit (Actinidia deliciosa)-derived actinidin, a cysteine protease, is renowned for its meat-tenderizing and milk-clotting activities. Despite its potential in various biotechnological applications, an efficient expression platform for actinidin production has not yet been developed. Instead, actinidin has traditionally been purified directly from the fruits of various plants. This study aimed to produce kiwifruit-derived actinidin in the leaves of Nicotiana benthamiana. The expressed actinidin was directed to the lumen of the endoplasmic reticulum (ER) using the binding immunoglobulin protein (BiP) signal sequence and an ER retention signal. To facilitate cost-effective purification, the family 3 cellulose-binding module (CBM3) was employed as an affinity tag, along with microcrystalline cellulose beads that bind efficiently to CBM3. A significant portion of the expressed actinidin was recovered in the soluble fraction without proteolytic degradation. The purified actinidin exhibited β-casein-degrading activity, with optimal efficiency observed at 55°C and pH 7.0. These results establish a promising plant-based platform for the efficient production and functional application of kiwifruit-derived actinidin in diverse biotechnological processes. |
| format | Article |
| id | doaj-art-e21b3c7bb9d2440491a2c7cd349e72c4 |
| institution | DOAJ |
| issn | 1664-462X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj-art-e21b3c7bb9d2440491a2c7cd349e72c42025-08-20T02:45:06ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2025-01-011510.3389/fpls.2024.15321701532170Expression of kiwifruit-derived actinidin in Nicotiana benthamiana leavesJi Hyun Kang0Jae-Ho Lee1Dong Wook Lee2Dong Wook Lee3Dong Wook Lee4Department of Integrative Food, Bioscience and Biotechnology, Chonnam National University, Gwangju, Republic of KoreaDepartment of Integrative Food, Bioscience and Biotechnology, Chonnam National University, Gwangju, Republic of KoreaDepartment of Integrative Food, Bioscience and Biotechnology, Chonnam National University, Gwangju, Republic of KoreaDepartment of Bioenergy Science and Technology, Chonnam National University, Gwangju, Republic of KoreaBio-Energy Research Center, Chonnam National University, Gwangju, Republic of KoreaKiwifruit (Actinidia deliciosa)-derived actinidin, a cysteine protease, is renowned for its meat-tenderizing and milk-clotting activities. Despite its potential in various biotechnological applications, an efficient expression platform for actinidin production has not yet been developed. Instead, actinidin has traditionally been purified directly from the fruits of various plants. This study aimed to produce kiwifruit-derived actinidin in the leaves of Nicotiana benthamiana. The expressed actinidin was directed to the lumen of the endoplasmic reticulum (ER) using the binding immunoglobulin protein (BiP) signal sequence and an ER retention signal. To facilitate cost-effective purification, the family 3 cellulose-binding module (CBM3) was employed as an affinity tag, along with microcrystalline cellulose beads that bind efficiently to CBM3. A significant portion of the expressed actinidin was recovered in the soluble fraction without proteolytic degradation. The purified actinidin exhibited β-casein-degrading activity, with optimal efficiency observed at 55°C and pH 7.0. These results establish a promising plant-based platform for the efficient production and functional application of kiwifruit-derived actinidin in diverse biotechnological processes.https://www.frontiersin.org/articles/10.3389/fpls.2024.1532170/fullactinidinmolecular farmingprotein expressionprotein purificationβ-casein degradation |
| spellingShingle | Ji Hyun Kang Jae-Ho Lee Dong Wook Lee Dong Wook Lee Dong Wook Lee Expression of kiwifruit-derived actinidin in Nicotiana benthamiana leaves Frontiers in Plant Science actinidin molecular farming protein expression protein purification β-casein degradation |
| title | Expression of kiwifruit-derived actinidin in Nicotiana benthamiana leaves |
| title_full | Expression of kiwifruit-derived actinidin in Nicotiana benthamiana leaves |
| title_fullStr | Expression of kiwifruit-derived actinidin in Nicotiana benthamiana leaves |
| title_full_unstemmed | Expression of kiwifruit-derived actinidin in Nicotiana benthamiana leaves |
| title_short | Expression of kiwifruit-derived actinidin in Nicotiana benthamiana leaves |
| title_sort | expression of kiwifruit derived actinidin in nicotiana benthamiana leaves |
| topic | actinidin molecular farming protein expression protein purification β-casein degradation |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2024.1532170/full |
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