Hyper-production of porcine contagious pleuropneumonia subunit vaccine proteins in Escherichia coli by developing a bicistronic T7 expression system
Author(s)
Type
Journal Article
Abstract
The ApxII toxin and the outer membrane lipoprotein (Oml) of Actinobacillus pleuropneumoniae are important vaccine antigens against porcine contagious pleuropneumonia (PCP), a prevalent infectious disease affecting the swine industry worldwide. Previous studies have reported the recombinant expression of ApxII and Oml in Escherichia coli; however, their yields were not satisfactory. Here, we aimed to enhance the production of ApxII and Oml by constructing a bicistronic expression system based on the widely used T7 promoter. To create efficient T7 bicistronic expression cassettes, 16 different fore-cistron sequences were introduced downstream of the T7 promoter. The expression of three vaccine antigens Oml1, Oml7, and ApxII in the four strongest bicistronic vectors were enhanced compared to the monocistronic control. Further optimization of the fermentation conditions in micro-well plates (MWP) led to improved production. Finally, the production yields reached unprecedented levels of 2.43 g L−1 of Oml1, 2.59 g L−1 of Oml7, and 1.21 g L−1 of ApxII, in a 5 L bioreactor. These three antigens also demonstrated well-protective immunity against A. pleuropneumoniae infection. In conclusion, this study establishes an efficient bicistronic T7 expression system that can be used to express recombinant proteins in E. coli and achieves the hyper-production of PCP vaccine proteins.
Date Issued
2024-01-01
Date Acceptance
2023-12-20
Citation
Biotechnology Journal, 2024, 19 (1)
ISSN
1860-6768
Publisher
Wiley
Journal / Book Title
Biotechnology Journal
Volume
19
Issue
1
Copyright Statement
Copyright © 2024 Wiley-VCH GmbH. This is the peer reviewed version of the following article: Sun, M., Gao, A. X., Li, A., Ledesma-Amaro, R., Wang, P., Chen, W., Bai, Z., & Liu, X. (2024). Hyper-production of porcine contagious pleuropneumonia subunit vaccine proteins in Escherichia coli by developing a bicistronic T7 expression system. Biotechnology Journal, 19, e2300187. https://doi.org/10.1002/biot.202300187, which has been published in final form at https://doi.org/10.1002/biot.202300187. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited.
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/38178735
Subjects
ACTINOBACILLUS-PLEUROPNEUMONIAE
bicistronic T7 expression system
Biochemical Research Methods
Biochemistry & Molecular Biology
Biotechnology & Applied Microbiology
DETERMINANTS
Escherichia coli
GENE-EXPRESSION
GENOME
hyper-production
Life Sciences & Biomedicine
OPTIMIZATION
OUTER-MEMBRANE LIPOPROTEIN
PATHOGENESIS
PCP subunit vaccine protein
recombinant expression
Science & Technology
STRATEGIES
TRANSLATION
VIRULENCE FACTORS
Publication Status
Published
Coverage Spatial
Germany
Article Number
2300187
Date Publish Online
2024-01-04