Abstract
Many venomous species, especially snakes, contain a variety of secreted phospholipases A2 that contribute to venom toxicity and prey digestion. We characterized a novel highly toxic phospholipase A2 of group II, WaPLA2-II, from the snake venom of Saudi Walterinnesia aegyptia (W. aegyptia). The enzyme was purified using a reverse phase C18 column. It is a monomeric protein with a molecular weight of approximately 14 kDa and an NH2-terminal amino acid sequence exhibiting similarity to the PLA2 group II enzymes. WaPLA2-II, which contains 2.5% (w/w) glycosylation, reached a maximal specific activity of 1250 U/mg at pH 9.5 and 55 °C in the presence of Ca2+ and bile salts. WaPLA2-II was also highly stable over a large pH and temperature range. A strong correlation between antimicrobial and indirect hemolytic activities of WaPLA2 was observed. Additionally, WaPLA2-II was found to be significantly cytotoxic only on cancerous cells. However, chemical modification with para-Bromophenacyl bromide (p-BPB) inhibited WaPLA2-II enzymatic activity without affecting its antitumor effect, suggesting the presence of a separate ‘pharmacological site’ in snake venom phospholipase A2 via its receptor binding affinity. This enzyme is a candidate for applications including the treatment of phospholipid-rich industrial effluents and for the food production industry. Furthermore, it may represent a new therapeutic lead molecule for treating cancer and microbial infections.
Highlights
The phospholipase A2 (PLA2) superfamily consists of many enzymes that catalyze the hydrolysis of the sn-2 ester bond in a variety of phospholipids
Peak 4, which was eluted at 32 min, attracted a lot our attention since it demonstrated good antibacterial and phospholipase activities
In the present study, the second active fraction (WaPLA2 -II) was characterized and compared to the previously reported WaPLA2-I. This single chromatographic step protocol demonstrated a significant degree of purity (25-fold) for WaPLA2 -II with a recovery yield of 11.9% based on the original PLA2 activity (Table 1)
Summary
Walterinnesia aegyptia Venom with Antimicrobial, Antifungal, and Cytotoxic Potential. Laboratory of Plant Biotechnology Applied to Crop Improvement, Faculty of Science of Sfax, University of. Received: 15 October 2020; Accepted: 23 November 2020; Published: 27 November 2020
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