Danger in the reef: Proteome, toxicity, and neutralization of the venom of the olive sea snake, Aipysurus laevis Artículo académico uri icon

Abstracto

  • Biodegradable polymers from renewable resources are generating growing interest in the plastic industry because they have properties similar to synthetic polymers. Polyhydroxyalkanoates, mainly polyhydroxybutyrate (PHB), have mechanical and physicochemical properties very similar to their synthetic counterparts. This work explores the use of residual glycerol from the bioenergy industry for the production of PHB by Bacillus megaterium DSM 32. The glycerol works as a source of carbon and energy. Raw glycerol was purified with sulfuric acid in order to neutralize saponified fatty acids. The purification process generated three different phases. One of the phases was the glycerol-rich layer; this layer was filtered and concentrated by vacuum distillation process. The purity of the glycerol was determined by thermogravimetric analysis (TGA). Additionally, the physicochemical properties, like viscosity, pH, ash content and density, were measured. The experiments were conducted in shake flasks at 30 °C and 120 rpm. Different glycerol concentrations (20, 30, 40 g/L) were used to evaluate the influence of the initial concentration of glycerol on the biomass accumulation and biopolymer production. The purified glycerol obtained had a high purity (∼ 89.5–92.13%); this material does not contain fatty acids, although it contains ∼3.7% salts. The final PHB concentration obtained was 0.054 mg/mL.
  • Four specimens of the olive sea snake, Aipysurus laevis, were collected off the coast of Western Australia, and the venom proteome was characterized and quantitatively estimated by RP-HPLC, SDS-PAGE, and MALDI-TOF-TOF analyses. A. laevis venom is remarkably simple and consists of phospholipases A2 (71.2%), three-finger toxins (3FTx; 25.3%), cysteine-rich secretory proteins (CRISP; 2.5%), and traces of a complement control module protein (CCM; 0.2%). Using a Toxicity Score, the most lethal components were determined to be short neurotoxins. Whole venom had an intravenous LD50 of 0.07 mg/kg in mice and showed a high phospholipase A2 activity, but no proteinase activity in vitro. Preclinical assessment of neutralization and ELISA immunoprofiling showed that BioCSL Sea Snake Antivenom was effective in cross-neutralizing A. laevis venom with an ED50 of 821 μg venom per mL antivenom, with a binding preference towards short neurotoxins, due to the high degree of conservation between short neurotoxins from A. laevis and Enhydrina schistosa venom. Our results point towards the possibility of developing recombinant antibodies or synthetic inhibitors against A. laevis venom due to its simplicity.

fecha de publicación

  • 2015
  • 2017