ORIGINAL RESEARCH article
Front. Plant Sci.
Sec. Plant Biotechnology
Volume 16 - 2025 | doi: 10.3389/fpls.2025.1631697
This article is part of the Research TopicPlant Molecular Farming for Biopharmaceutical Production and BeyondView all 8 articles
Transient Expression of Full-Length and Mature Nattokinase in Nicotiana benthamiana Reveals Early Necrosis from Full-Length Form and Functional Activity of the Mature Enzyme
Provisionally accepted
- 1University of Pikeville, Pikeville, United States
- 2Arizona State University, Tempe, Arizona, United States
- 3Chinese Academy of Agricultural Sciences (CAAS), Beijing, Beijing Municipality, China
Select one of your emails
You have multiple emails registered with Frontiers:
Notify me on publication
Please enter your email address:
If you already have an account, please login
You don't have a Frontiers account ? You can register here
Nattokinase is a potent fibrinolytic enzyme widely used in the treatment of cardiovascular diseases for its ability to directly degrade fibrin and plasmin substrates, effectively dissolving blood clots. In this study, both full-length and mature forms of the nattokinase coding sequence were transiently expressed in Nicotiana benthamiana using a modified Bean Yellow Dwarf Virus (BeYDV) replicon system. Overexpression of the full-length (pre-pro) construct resulted in severe leaf necrosis within 2.5 days post-infiltration (dpi), with electrolyte leakage analysis indicating an 83.5% loss of membrane integrity by 3 dpi. In contrast, the mature form of nattokinase was successfully expressed without early cytotoxicity and exhibited strong caseinolytic and fibrinolytic activity, reaching 22,500 FU/g-comparable to commercial standards. These findings demonstrate the feasibility of producing biologically active nattokinase in plants and highlight the potential of plant-based expression systems as scalable, cost-effective platforms for therapeutic enzyme production.
Keywords: Plant transient expression, codon optimization, Nattokinase, BeYDV, Nicotiana benthamiana
Received: 20 May 2025; Accepted: 30 Jun 2025.
Copyright: © 2025 Wang, Mason, Hall, Slone, Tackett and Wang. 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:
Kevin Yueju Wang, University of Pikeville, Pikeville, United States
Nan Wang, Chinese Academy of Agricultural Sciences (CAAS), Beijing, 100081, Beijing Municipality, China
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.