A method for efficient expression of Pseudomonas aeruginosa alginate lyase in Pichia pastoris
| Autor: | Yu Qiao, Michael M Yue, Hongbiao Ding, Wendy W Gong |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Genetic Vectors 030106 microbiology Peptide Biology Protein Engineering medicine.disease_cause Biochemistry Pichia law.invention Microbiology Pichia pastoris 03 medical and health sciences Hydrolysis Bacterial Proteins law medicine Cloning Molecular Polysaccharide-Lyases chemistry.chemical_classification Pseudomonas aeruginosa Kanamycin General Medicine Antimicrobial biology.organism_classification Recombinant Proteins Anti-Bacterial Agents 030104 developmental biology Enzyme chemistry Recombinant DNA Biotechnology medicine.drug |
| Zdroj: | Preparative Biochemistry & Biotechnology. 46:165-170 |
| ISSN: | 1532-2297 1082-6068 |
| DOI: | 10.1080/10826068.2014.996233 |
| Popis: | As an eco-friendly biocatalyst for alginate hydrolysis, bacteria-derived alginate lyase (AlgL) has been widely used in research and industries to produce oligosaccharides. However, the cost of AlgL enzyme production remains high due to the low expression and difficulty in purification from bacterial cells. In this study we report an effective method to overexpress the Pseudomonas aeruginosa AlgL (paAlgL) enzyme in Pichia pastoris. Fused with a secretory peptide, the recombinant paAlgL was expressed extracellularly and purified from the culture supernatant through a simple process. The purified recombinant enzyme is highly specific for alginate sodium with a maximal activity of 2,440 U/mg. The enzymatic activity remained stable below 45°C and at pH between 4 and 10. The recombinant paAlgL was inhibited by Zn(2+), Cu(2+), and Fe(2+) and promoted by Co(2+) and Ca(2+). Interestingly, we also found that the recombinant paAlgL significantly enhanced the antimicrobial activity of antibiotics ampicillin and kanamycin against Pseudomonas aeruginosa. Our results introduce a method for efficient AlgL production, the characterization, and a new feature of the recombinant paAlgL as an enhancer of antibiotics against Pseudomonas aeruginosa. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|