Skip to main content

BRIEF RESEARCH REPORT article

Front. Plant Sci.
Sec. Plant Biotechnology
Volume 15 - 2024 | doi: 10.3389/fpls.2024.1532170

Expression of kiwifruit-derived actinidin in Nicotiana benthamiana leaves

Provisionally accepted
Ji Hyun Kang Ji Hyun Kang Jae Ho Lee Jae Ho Lee Dong Wook Lee Dong Wook Lee *
  • Chonnam National University, Gwangju, Republic of Korea

The final, formatted version of the article will be published soon.

    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-caseindegrading 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.

    Keywords: Actinidin, Molecular Farming, protein expression, Protein purification, β-casein degradation

    Received: 21 Nov 2024; Accepted: 11 Dec 2024.

    Copyright: © 2024 Kang, Lee and Lee. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Dong Wook Lee, Chonnam National University, Gwangju, Republic of Korea

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.