The final, formatted version of the article will be published soon.
METHODS article
Front. Bioeng. Biotechnol.
Sec. Industrial Biotechnology
Volume 12 - 2024 |
doi: 10.3389/fbioe.2024.1436352
Production and Purification of Outer Membrane Vesicles (OMVs) Encapsulating Green Fluorescent Protein from Escherichia coli: A Step towards Scalable OMV Technologies
Provisionally accepted- 1 University of Los Andes, Colombia, Bogotá, Colombia
- 2 Universidad Antonio Nariño, Bogotá, Cundinamarca, Colombia
Outer membrane vesicles (OMVs) are spherical structures that contain a small fraction of the periplasm of Gram-negative bacteria, surrounded by its outer membrane. They are naturally produced and detached from the bacterial surface, participate in diverse biological processes, and their diameter size is in the range of 10–300 nm. OMVs have gained interest in different applications, such as the development of biosensors, vaccines, protein chips, and the encapsulation of heterologous proteins and peptides expressed by these microorganisms. However, the use of OMVs in these applications is limited due to the low yields and high purification costs. In this study, we produced green fluorescent protein (GFP) encapsulated into OMVs using Escherichia coli JC8031 transformed with pTRC99A-ssTorA-GFP to establish the production and purification route. Results showed that the motility of the strain prevents its immobilization in alginate, which hampers the purification of OMVs. To address this issue, a zeolite-based column was used to chromatographically separate the OMVs from smaller particles. Further experiments will be focused on standardizing the production and purification of OMVs at a scalable level.
Keywords: outer membrane vesicles, green fluorescent protein, protein encapsulation, Cell Immobilization, size exclusion chromatography (SEC)
Received: 21 May 2024; Accepted: 04 Nov 2024.
Copyright: © 2024 Torres-Vanegas, Rincon-Tellez, Guzmán-Sastoque, Valderrama-Rincon, Cruz and Reyes. 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:
Juan C. Cruz, University of Los Andes, Colombia, Bogotá, Colombia
Luis H. Reyes, University of Los Andes, Colombia, Bogotá, Colombia
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.