ArticleBMC biotechnology2015
Soluble expression, purification, and characterization of active recombinant human tissue plasminogen activator by auto-induction in E. coli.
Article in BMC biotechnology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 31 citations in OpenAlex.
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- Design of Experiments As a Tool for Optimization in Recombinant Protein Biotechnology: From Constructs to Crystals.Molecular biotechnology · 2019Review
- Improvement of solubility and yield of recombinant protein expression inResearch in pharmaceutical sciences · 2019Article
- Two Simple Methods for Optimizing the Production of "Difficult-to-Express" GnRH-DFF40 Chimeric Protein.Advanced pharmaceutical bulletin · 2019Article
- High-yield Production of Granulocyte-macrophage Colony-stimulating Factor inIranian journal of pharmaceutical research : IJPR · 2019Article
- Reteplase: Structure, Function, and Production.Advanced biomedical research · 2019Review
- Structure-based antigenic epitope and PEGylation improve the efficacy of staphylokinase.Microbial cell factories · 2017Article
- Functionally Expression of Metalloproteinase in Taenia solium Metacestode and Its Evaluation for Serodiagnosis of Cysticercosis.Iranian journal of parasitologyArticle
- Optimization of the Production of Soluble Recombinant TEV Protease in TwoAvicenna journal of medical biotechnologyArticle
- Comparison of Three Escherichia coli Strains in Recombinant Production of Reteplase.Avicenna journal of medical biotechnologyArticle
Corrections and comments
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Authors and funding
12 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundHuman tissue plasminogen activator (tPA) belongs to the serine protease family. It converts plasminogen into plasmin and is used clinically to treat thrombosis. Human tPA is composed of 527 amino acids residues and contains 17 disulfide bonds. Escherichia coli has been used only rarely for the efficient production of recombinant tPA. However, the functional expression of full-length tPA that contains multiple disulfide bonds on an industrial scale remains challenging. Here, we describe the soluble expression and characterization of full-length tPA by auto-induction in E. coli.
resultsWe achieved optimal levels of gene expression, minimized negative effects related to the production of heterologous proteins, and optimized cytoplasmic yields. Three different E. coli strains, BL21 (DE3), Rosetta, and Origami 2, could express tPA using an auto-induction mechanism. In addition, similar yields of recombinant protein were produced at temperatures of 33, 35, and 37°C. The E. coli strain origami 2 could increase disulfide bond formation in cytoplasmic tPA and produce purified soluble recombinant protein (~0.9 mg/l medium). The full-length tPA was monomeric in solution, and fibrin plate assays confirmed that the recombinant tPA displayed serine protease activity.
conclusionsThis is the first report that describes the heterologous expression of correctly folded active full-length tPA. This could provide valuable information for using prokaryotic auto-induction expression systems to produce tPA at industrial and pharmaceutical levels without in vitro refolding during the production step.
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