Production of a modified peptide clavanin in Pichia pastoris: cloning, expression, purification and in vitro activities
| Autor: | Fábio Correa Carneiro, Octavio L. Franco, Priscilla Santos Aguiar, Simoni Campos Dias, Nádia Skorupa Parachin, Kelly C.L. Mulder, Loiane Alves de Lima |
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| Jazyk: | angličtina |
| Předmět: |
chemistry.chemical_classification
Antimicrobial peptides Biophysics Heterologous Peptide Clavanin Biology biology.organism_classification Applied Microbiology and Biotechnology Yeast In vitro Pichia pastoris Microbiology Biochemistry chemistry Original Article Heterologous expression Thioredoxin Antimicrobial peptide |
| Zdroj: | AMB Express |
| ISSN: | 2191-0855 |
| DOI: | 10.1186/s13568-015-0129-0 |
| Popis: | Antimicrobial peptides are one of the most promising peptide-based drugs due to their enormous potential as novel biopharmaceuticals in both human and animal industries. In order to develop strategies to over produce such molecules, heterologous production of a modified version of clavanin A, here named clavanin MO (clavMO), was successfully achieved in the methylothopic yeast Pichia pastoris. ClavMO was fused to thioredoxin as a carrier protein and the construction was tested using two promoters, PAOX1 and PGAP, based on either induced or constitutive expression systems, respectively. After growth in 5 L Bioreactor, clavMO-thio was recovered and purified through size exclusion chromatography. Our findings show that both constitutive and inducible expression systems produce active clavMO fused to thioredoxin against both Gram-negative Klebsiella pneumoniae and Gram-positive Staphylococcus aureus microorganisms. Electronic supplementary material The online version of this article (doi:10.1186/s13568-015-0129-0) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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