ArticleMicrobial cell factories2017
Large-scale production of a thermostable Rhodothermus marinus cellulase by heterologous secretion from Streptomyces lividans.
Article in Microbial cell factories, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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16 citing papers in PubMed, 49 citations in OpenAlex.
- Multidimensional strategy enables scalable metabolome diversity in microbial fermentations.Scientific reports · 2026Article
- Bacillus and Streptomyces spp. as hosts for production of industrially relevant enzymes.Applied microbiology and biotechnology · 2024Review
- Enhanced protein secretion in reduced genome strains of Streptomyces lividans.Microbial cell factories · 2024Article
- Secretory expression of recombinant small laccase genes in Gram-positive bacteria.Microbial cell factories · 2023Article
- Development of a thiostrepton-free system for stable production of PLD in Streptomyces lividans SBT5.Microbial cell factories · 2022Article
- Droplet-based microfluidic platform for high-throughput screening of Streptomyces.Communications biology · 2021Article
- Review
- A contradictory action of procoagulant ficin by a fibrinolytic serine protease from EgyptianBiotechnology reports (Amsterdam, Netherlands) · 2020Article
- Streptomycetes: Attractive Hosts for Recombinant Protein Production.Frontiers in microbiology · 2020Review
- A novel autolysis system for extracellular production and direct immobilization of a phospholipase D fused with cellulose binding domain.BMC biotechnology · 2019Article
- Secretome Dynamics in a Gram-Positive Bacterial Model.Molecular & cellular proteomics : MCP · 2019Article
- The Transcription Unit Architecture ofFrontiers in microbiology · 2019Article
- Transcriptomic and fluxomic changes in Streptomyces lividans producing heterologous protein.Microbial cell factories · 2018Article
- Comprehensive subcellular topologies of polypeptides in Streptomyces.Microbial cell factories · 2018Article
- Monitoring Protein Secretion inFrontiers in microbiology · 2018Article
- Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases inFrontiers in microbiology · 2018Article
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Authors and funding
13 authors at 3 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe gene encoding a thermostable cellulase of family 12 was previously isolated from a Rhodothermus marinus through functional screening. CelA is a protein of 260 aminoacyl residues with a 28-residue amino-terminal signal peptide. Mature CelA was poorly synthesized in some Escherichia coli strains and not at all in others. Here we present an alternative approach for its heterologous production as a secreted polypeptide in Streptomyces.
resultsCelA was successfully over-expressed as a secreted polypeptide in Streptomyces lividans TK24. To this end, CelA was fused C-terminally to the secretory signal peptide of the subtilisin inhibitor protein (Sianidis et al. in J Biotechnol. 121: 498-507, 2006) from Streptomyces venezuelae and a new cloning strategy developed. Optimal growth media and conditions that stall biomass production promote excessive CelA secretion. Under optimal growth conditions in nutrient broth medium, significant amounts of mature CelA (50-90 mg/L or 100-120 mg/g of dry cell weight) are secreted in the spent growth media after 7 days. A protocol to rapidly purify CelA to homogeneity from culture supernatants was developed and specific anti-sera raised against it. Biophysical, biochemical and immmuno-detection analyses indicate that the enzyme is intact, stable and fully functional. CelA is the most thermostable heterologous polypeptide shown to be secreted from S. lividans.
conclusionThis study further validates and extends the use of the S. lividans platform for production of heterologous enzymes of industrial importance and extends it to active thermostable enzymes. This study contributes to developing a platform for poly-omics analysis of protein secretion in S. lividans.
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