Recombinant expression and anti-microbial activity of anti-lipopolysaccharide factor (ALF) from the black tiger shrimp
| Autor: | Evelyne Bachère, Yannick Gueguen, Sirawut Klinbunga, Bernard Romestand, Michael Marcos, Kunlaya Somboonwiwat, Guy Moulin, Anchalee Tassanakajon, André Aumelas, Hélène Boze |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
0303 health sciences
biology Immunology 04 agricultural and veterinary sciences biology.organism_classification Vibrio Yeast Shrimp Microbiology law.invention Penaeus monodon Pichia pastoris 03 medical and health sciences Transformation (genetics) law 040102 fisheries Recombinant DNA 0401 agriculture forestry and fisheries 14. Life underwater Bacteria 030304 developmental biology Developmental Biology |
| Zdroj: | Developmental & Comparative Immunology. 29:841-851 |
| ISSN: | 0145-305X |
| Popis: | Anti-lipopolysaccharide factors (ALFs), originally characterized from horseshoe crabs, have been recently identified from hemocytes of the black tiger shrimp, Penaeus monodon, by a genomic approach. In order to characterize the properties and biological activities of this immune effector in shrimp, ALFPm3, the most abundant isoform found in P. monodon, was expressed in the yeast Pichia pastoris. Large-scale production in fermentor provided 262 mg/l of recombinant ALFPm3 which was purified to homogeneity by single chromatography step on expanded-bed Streamline SP6XL. The rALFPm3 was further characterized in terms of N-terminal sequencing and mass spectrometry. Anti-microbial assays demonstrated that rALFPm3 has a broad spectrum of anti-fungal properties against filamentous fungi, and anti-bacterial activities against both Gram-positive and Gram-negative bacteria, associated with a bactericidal effect. Interestingly, rALFPm3 is highly efficient against various Vibrio species including strains pathogenic for shrimp. Finally, a synthetic peptide corresponding to a part of the putative LPS-binding site of ALFPm3 was shown to display activities mainly directed against Gram-positive bacteria indicating the involvement of the full molecule to the anti-microbial activity for Gram-negative bacteria. |
| Databáze: | OpenAIRE |
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