ArticleMicrobial cell factories2014
Double mutation of cell wall proteins CspB and PBP1a increases secretion of the antibody Fab fragment from Corynebacterium glutamicum.
Article in Microbial cell factories, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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20 citing papers in PubMed, 61 citations in OpenAlex.
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- The pH-responsive precipitation-redissolution of the CspB fusion protein, CspB50TEV-Teriparatide, triggered by changes in secondary structure.Biochemistry and biophysics reports · 2023Article
- Efficient secretory production of proline/alanine/serine (PAS) biopolymers in Corynebacterium glutamicum yielding a monodisperse biological alternative to polyethylene glycol (PEG).Microbial cell factories · 2022Article
- Hypersecretion of OmlA antigen in Corynebacterium glutamicum through high-throughput based development process.Applied microbiology and biotechnology · 2022Article
- Application of Corynebacterium glutamicum engineering display system in three generations of biorefinery.Microbial cell factories · 2022Review
- Single-chain tandem macrocyclic peptides as a scaffold for growth factor and cytokine mimetics.Communications biology · 2022Article
- Study on Demethoxycurcumin as a Promising Approach to Reverse Methicillin-Resistance ofInternational journal of molecular sciences · 2021Article
- PASylated Thymosin α1: A Long-Acting Immunostimulatory Peptide for Applications in Oncology and Virology.International journal of molecular sciences · 2020Article
- Enhanced production of recombinant proteins in Corynebacterium glutamicum by constructing a bicistronic gene expression system.Microbial cell factories · 2020Article
- Development of a secretory expression system with high compatibility between expression elements and an optimized host for endoxylanase production in Corynebacterium glutamicum.Microbial cell factories · 2019Article
- Triple deletion of clpC, porB, and mepA enhances production of small ubiquitin-like modifier-N-terminal pro-brain natriuretic peptide in Corynebacterium glutamicum.Journal of industrial microbiology & biotechnology · 2019Article
- Recombinant Protein Expression System inFrontiers in microbiology · 2018Review
- Polynucleotide Phosphorylase, RNase E/G, and YbeY Are Involved in the Maturation of 4.5S RNA in Corynebacterium glutamicum.Journal of bacteriology · 2017Article
- Comparative analysis of Corynebacterium glutamicum genomes: a new perspective for the industrial production of amino acids.BMC genomics · 2017Article
- Transcriptome analysis of Corynebacterium glutamicum in the process of recombinant protein expression in bioreactors.PloS one · 2017Article
- Use of a Sec signal peptide library from Bacillus subtilis for the optimization of cutinase secretion in Corynebacterium glutamicum.Microbial cell factories · 2016Article
- Corynebacterium glutamicum possesses β-N-acetylglucosaminidase.BMC microbiology · 2016Article
- Enhancing the selective extracellular location of a recombinant E. coli domain antibody by management of fermentation conditions.Applied microbiology and biotechnology · 2015Article
- A chromosomally encoded T7 RNA polymerase-dependent gene expression system for Corynebacterium glutamicum: construction and comparative evaluation at the single-cell level.Microbial biotechnology · 2015Article
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9 authors at 4 institutions in 2 countries.
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Abstract
backgroundAmong other advantages, recombinant antibody-binding fragments (Fabs) hold great clinical and commercial potential, owing to their efficient tissue penetration compared to that of full-length IgGs. Although production of recombinant Fab using microbial expression systems has been reported, yields of active Fab have not been satisfactory. We recently developed the Corynebacterium glutamicum protein expression system (CORYNEX®) and demonstrated improved yield and purity for some applications, although the system has not been applied to Fab production.
resultsThe Fab fragment of human anti-HER2 was successfully secreted by the CORYNEX® system using the conventional C. glutamicum strain YDK010, but the productivity was very low. To improve the secretion efficiency, we investigated the effects of deleting cell wall-related genes. Fab secretion was increased 5.2 times by deletion of pbp1a, encoding one of the penicillin-binding proteins (PBP1a), mediating cell wall peptidoglycan (PG) synthesis. However, this Δpbp1a mutation did not improve Fab secretion in the wild-type ATCC13869 strain. Because YDK010 carries a mutation in the cspB gene encoding a surface (S)-layer protein, we evaluated the effect of ΔcspB mutation on Fab secretion from ATCC13869. The Δpbp1a mutation showed a positive effect on Fab secretion only in combination with the ΔcspB mutation. The ΔcspBΔpbp1a double mutant showed much greater sensitivity to lysozyme than either single mutant or the wild-type strain, suggesting that these mutations reduced cell wall resistance to protein secretion.
conclusionThere are at least two crucial permeability barriers to Fab secretion in the cell surface structure of C. glutamicum, the PG layer, and the S-layer. The ΔcspBΔpbp1a double mutant allows efficient Fab production using the CORYNEX® system.
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