Aim: This study aimed to investigate the antiviral activity of Pterois volitans phospholipase A2 (PV-PLA2) from Indonesia to human immunodeficiency virus (HIV).
Materials and Methods: Fresh venomous fin parts of wild PV specimens were collected from Java Sea waters. Then, it washed using phosphate buffer pH 7.0 and immersed in phosphate buffer pH 7.0 0.01 m containing CaCl2 0.001 m for 24 h. The immersed fin then allowed for extraction process by sonicating for 2×8 min with 80% pulse and 20 kHz output with temperature controlling to avoid denaturation. The crude venom (CV) extracted from the fin is allowed for purification by 80% ethanol (ET) precipitation and ammonium sulfate fractionation method. The purified PV-PLA2 then analyzed using Lowry's method, Marinette's method, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 3-(4, 5-dimethyl thiazol-2yl)-2, 5-diphenyl tetrazolium bromide assay. After determining the purest and safest sample of six samples analyzed, the chosen sample then tested into simian retrovirus-2 (SRV2)-A549 culture (48×104 cells/mL at 1-4 ppm), and compared to the CV sample (1-4 ppm) and lamivudine (100 ppm). The culture then is analyzed using a quantitative real time-polymerase chain reaction to find out the copy number of SRV-2 virus in each culture.
Results: The protein's activity, concentration, and purity analysis revealed that the PV-PLA2 purified using ammonium sulfate fractionation has the highest activity (1.81 times higher than the CV at 80% fractionation) and has higher purity than the sample from ET fractionation. The testing of the sample purified using ammonium sulfate fractionation at 80% saturation level shown that it has a 97.78% inhibition level toward SRV2-A549 culture at 4 ppm. However, in comparison to lamivudine which has 99.55% inhibition level at 100 ppm, it needs much lower concentration to achieve the same result.
Conclusion: The significant inhibition of SRV2-A549 culture shown that the PV-PLA2 extracted from PV venom has the potential to become anti-HIV substances. It would be worthwhile to further evaluate the antiretroviral activity of PV-PLA2 in the in vivo studies.
Pterois volitans, or commonly referred to lionfish, are fish species originating from Indo-Pacific waters but are becoming invasive in other regions such as the Caribbean and Atlantis. Various efforts have been made to reduce the number of lionfish, and one of them is by utilizing the venom on the spine. The venom extraction of P. volitans spines is done mechanically using sonication and centrifugation, and then protein isolation is carried out using salt. Coagulant activity from extract (crude venom) and lionfish venom protein isolate was done by counting PT (prothrombin time) and aPTT (activated partial thromboplastin time) which resulted that the crude venom and protein isolate of lionfish venom can accelerate blood clot (procoagulant) respectively up to 8.5 seconds and 6 seconds. Protein identification was made using LC-MS/MS device. The LC-MS/MS analysis showed that the protein isolate of lionfish venom contains Nomega-nitro-L-arginine methyl ester (L-NAME) compounds known to have procoagulant effects. From a series of tests mentioned, it concluded that P. volitans venom have procoagulant activity and one of the compounds responsible for it is L-NAME
The population of Pterois volitans has caused significant damage to other fish populations and coral reef ecosystems. Population control of P. volitans consumes a considerable cost so that the utilization of these fish needs to be sought to be useful along with controlling the population. This fish is known to contain the enzyme Phospholipase A2 (PLA2), which used as an antibiotic against some bacteria. This study will examine the antibacterial activity of the phospholipase A2 enzyme extracted from P. volitans venom to Escherichia coli, Bacillus subtilis, and Staphylococcus aureus. The method used to isolate the enzyme PLA2 is by using precipitation of ammonium sulfate (AS) and precipitation with ethanol. The results of the precipitation tested with the Lowry protein concentration test, the Marinetti PLA2 activity test, and the identification of the SDS-PAGE protein. The agar diffusion disc method is used to test the antibacterial activity. The results obtained from this research are that 80% ammonium sulfate precipitation method has the highest protein and enzyme activity with a ratio of 1.32 times compared to toxic extract. For antibacterial activity test results, an 80% ammonium sulfate sample may inhibit the activity of S. aureus bacteria but does not affect B. subtilis and E. coli.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.