ArticleMicrobial cell factories2020
Enhanced production of recombinant proteins in Corynebacterium glutamicum by constructing a bicistronic gene expression system.
Article in Microbial cell factories, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 34 citations in OpenAlex.
- Engineering genetic elements for microbial protein expression systems: Advances, challenges, applications, and prospects.Synthetic and systems biotechnology · 2026Review
- Rapid Identification of Superior Endogenous Signal Peptides for Heterologous Protein Secretion by Corynebacterium glutamicum Through Modular Cloning and Automation.Microbial biotechnology · 2026Article
- Engineered membraneless organelles inSynthetic and systems biotechnology · 2025Article
- Review of the Proteomics and Metabolic Properties ofMicroorganisms · 2024Review
- Establishment of CRISPR-Cpf1-assisted gene editing tool and engineering of 4-hydroxyisoleucine biosynthesis in Corynebacterium glutamicum.World journal of microbiology & biotechnology · 2023Article
- Leaderless Bicistronic Design for Precise and Reliable Control of Gene Expression inACS synthetic biology · 2023Article
- High-level recombinant protein production with Corynebacterium glutamicum using acetate as carbon source.Microbial biotechnology · 2022Article
- Development of a novel platform for recombinant protein production inSynthetic and systems biotechnology · 2022Article
- Article
- Construction of an ultra-strong PtacM promoter via engineering the core-element spacer and 5' untranslated region for versatile applications inBiotechnology notes (Amsterdam, Netherlands) · 2022Article
- Importance of the 5' regulatory region to bacterial synthetic biology applications.Microbial biotechnology · 2021Review
- Development of a Hyperosmotic Stress Inducible Gene Expression System by Engineering the MtrA/MtrB-DependentFrontiers in microbiology · 2021Article
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Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
Abstract
backgroundCorynebacterium glutamicum is a traditional food-grade industrial microorganism, in which an efficient endotoxin-free recombinant protein expression factory is under developing in recent years. However, the intrinsic disadvantage of low recombinant protein expression level is still difficult to be solved. Here, according to the bacteria-specific polycistronic feature that multiple proteins can be translated in one mRNA, efforts have been made to insert a leading peptide gene upstream of target genes as an expression enhancer, and it is found that this can remarkably improve the expression level of proteins under the control of inducible tac promoter in C. glutamicum.
resultsIn this research, the Escherichia coli (E. coli) tac promoter combined with 24 different fore-cistron sequences were constructed in a bicistronic manner in C. glutamicum. Three strong bicistronic expression vectors were isolated and exhibited high efficiency under different culture conditions. The compatibility of these bicistronic vectors was further validated using six model proteins- aldehyde dehydrogenase (ALDH), alcohol dehydrogenase (ADH), RamA (regulator of acetate metabolism), Bovine interferon-α (BoIFN-α), glycoprotein D protein (gD) of infectious bovine rhinotracheitis virus (IBRV) and procollagen type Ι N-terminal peptide (PΙNP). All examined proteins were highly expressed compared with the original vector with tac promoter. Large-scale production of PΙNP was also performed in fed-batch cultivation, and the highest PΙNP production level was 1.2 g/L.
conclusionIn this study, the strength of the inducible tac promoter for C. glutamicum was improved by screening and inserting fore-cistron sequences in front of the target genes. Those vectors with bicistronic expression patterns have strong compatibility for expressing various heterogeneous proteins in high yield. This new strategy could be used to further improve the performance of inducible promoters, achieving double competence of inducible control and high yield.
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