Effects of Jatropha Curcas and Acacia Nilotica Plants Extract Phenolics on the Activity of Partially Purified Phospholipase A2 Enzyme of Naja Katiensis Snake Venom

Hassan Abba Umar, Nkechi Eucharia Egbe, Ugochukwu Okechukwu Ozojiofor, Onuh Kingsley, Kereakede Ebipade, Umar Sani Inuwa, Madu Adulkarim Him, Fatima Abdullahi Harun

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Volume 1 | Issue 1 | Year 2025 | Article Id: RLS-V1I1P101

Effects of Jatropha Curcas and Acacia Nilotica Plants Extract Phenolics on the Activity of Partially Purified Phospholipase A2 Enzyme of Naja Katiensis Snake Venom

Hassan Abba Umar, Nkechi Eucharia Egbe, Ugochukwu Okechukwu Ozojiofor, Onuh Kingsley, Kereakede Ebipade, Umar Sani Inuwa, Madu Adulkarim Him, Fatima Abdullahi Harun

Received	Revised	Accepted	Published
02 Apr 2025	03 May 2025	02 Jun 2025	30 Jun 2025

Citation

Hassan Abba Umar, Nkechi Eucharia Egbe, Ugochukwu Okechukwu Ozojiofor, Onuh Kingsley, Kereakede Ebipade, Umar Sani Inuwa, Madu Adulkarim Him, Fatima Abdullahi Harun . “Effects of Jatropha Curcas and Acacia Nilotica Plants Extract Phenolics on the Activity of Partially Purified Phospholipase A2 Enzyme of Naja Katiensis Snake Venom.” DS Reviews of Research in Life Sciences, vol. 1, no. 1, pp. 1-12, 2025.

Abstract

Snake envenomation issues have been for long a neglected public health problem in Nigeria and other West African countries, causing a high number of human fatalities yearly. It is, however, still a serious social, pharmaceutical, economic and medical issue. This experimental research is aimed at the biocharacterization of the partially purified Phospholipase A2 enzyme of Naja katiensis venom. The venom enzyme is extracted using a two-way purification step (Gel filtration using G-75 and ion exchange chromatography on CM-Sephadex). The molecular weight of the enzyme is determined using Sodium Dodecyl-Sulphate Electrophoresis. The extracted enzyme kinetic parameters were also determined, after which its relative optimum affinity/activity was also determined related to different temperature ranges, pH, metal ions and salts. The partially purified PLA2 gave a total enzyme activity of 3.11 µmol/min, with an estimated molecular weight of 17.5 KDa. Initial velocity data of the enzyme was used to compute the kinetic velocity of the enzyme, where the Km and Vmax of the enzyme were estimated to be 12.6 mg/ml and 3.32 μmoles/min, respectively. The enzyme’s optimum temperature and pH were found to be 35oC and 7.0. Ca2+ ions were revealed to increase enzyme activity and affinity. Enzyme inhibition analyses done for the two plants’ phenolic fractions reveal that the two plant fractions have some level of inhibitory capability against the snake venom PLA2 enzyme. The research revealed that data could provide an alternative natural way of producing a more friendly pharmaceutical formulation for managing and treating snake envenomation.

Keywords

Antiserum, Chromatography, Envenomation, Kinetic, Naja Katiensis, Venom.

References

[1] Ashis K. Mukherjee, and Stephen P. Mackessy, “Prevention and Improvement of Clinical Management of Snakebite in Southern Asian countries: A Proposed Road Map,” Toxicon, vol. 200, pp. 140−152, 2021.

[CrossRef] [Google Scholar] [Publisher Link]

[2] Abdulrazaq G. Habib, “Public Health Aspects of Snakebite Care in West Africa: Perspectives from Nigeria,” Journal of Venomous Animals and Toxins Including Tropical Diseases, vol. 27, 2013.

[CrossRef] [Google Scholar] [Publisher Link]

[3] Aleksandra Bocian, and Konrad K. Hus, “Antibacterial Properties of Snake Venom Components,” Chemical Papers, vol. 74, pp. 407–419, 2020.

[CrossRef] [Google Scholar] [Publisher Link]

[4] Marianne Molander et al., “Hyaluronidase, phospholipase A2 and protease inhibitory activity of plants used in traditional treatment of snakebite-induced tissue necrosis in Mali, DR Congo and South Africa,” Journal of Ethnopharmacology, vol. 157, pp. 171-180, 2014.

[CrossRef] [Google Scholar] [Publisher Link]

[5] Tse Siang Kang et al., “Enzymatic Toxins from Snake Venom: Structural Characterization and Mechanism of Catalysis,” The FEBS Journal, vol. 278, no. 23, pp. 4544-4576, 2011.

[CrossRef] [Google Scholar] [Publisher Link]

[6] Mohammad Fahim Kadir et al., “Ethnopharmacological Survey of Medicinal Plants Used by Traditional Healers and Indigenous People in Chittagong Hill Tracts, Bangladesh, for the Treatment of Snakebite,” Evidence-Based Complementary and Alternative Medicine, 2015.

[CrossRef] [Google Scholar] [Publisher Link]

[7] Islem Abid et al., “New Group II Phospholipase A₂ from Walterinnesia Aegyptia Venom with Antimicrobial, Antifungal and Cytotoxic Potential,” Processes, vol. 8, no. 12, 2020.

[CrossRef] [Google Scholar] [Publisher Link]

[8] A.B. Sallau et al., “Characterization of Phospholipase A₂ from Echis Ocellatus Venom,” African Journal of Biochemistry Research, vol. 2, no. 4, pp. 98-101, 2008.

[Google Scholar] [Publisher Link]

[9] B.G. Kurfi, M.A. Abdulazeez, and Z. Tukur, “Effects of Increased Creatine Kinase Levels in Rabbits Due to PLA₂ of Naja Nigricullis Venom,” Bayero Journal BioMedical Science, vol. 1, no. 1, pp. 49-54, 2016.

[Publisher Link]

[10] Nasio A. Nasio, “Snake Bites: A Forgotten Menace in Kenya,” International Journal of Scientific Research and Innovative Technology, vol. 3, no. 7, 2016.

[Google Scholar]

[11] Victor Pecina Quintero et al., “Genetic Structure of Jatropha Curcas L. in Mexico and Probable Centre of Origin,” Biomass and Bioenergy, vol. 60, pp. 147-155, 2014.

[CrossRef] [Google Scholar] [Publisher Link]

[12] Gamal Fakhry et al., “Use of Waste Water and Treated Water for Jatropha Curcas Cultivation and the Possibility of Oil Seed Use as a Biofuel,” Minia Journal of Agricultural Research & Development, vol. 36, no. 2, pp. 245-269, 2016.

[Google Scholar] [Publisher Link]

[13] T. Kalaivani, and Lazar Mathew, “Free Radical Scavenging Activity from Leaves of Acacia Nilotica (L.) Wil. Ex-Delile, an Indian Medicinal Tree,” Food and Chemical Toxicology, vol. 48, no. 1, pp. 298-305, 2010.

[CrossRef] [Google Scholar] [Publisher Link]

[14] A.A. Jigam et al., “Polygalloyltannin Isolated from the Roots of Acacia Nilotica Del. (Leguminoseae) is Effective against Plasmodium Berghei in Mice,” Journal of Medicinal Plants Research, vol. 4, no. 12, pp. 1169-1175, 2010.

[Google Scholar] [Publisher Link]

[15] A. Kumar et al., “Phytochemical Investigation on Tropical Plant,” Pakistan Journal of Nutrition, vol. 8, no. 1, pp. 83-85, 2009.

[Google Scholar] [Publisher Link]

[16] Abayomi Sofowora, “Research on Medicinal Plants and Traditional Medicinal in Africa,” The Journal of Alternative and Complementary Medicine, vol. 2, no. 3, pp. 365-372, 1996.

[CrossRef] [Google Scholar] [Publisher Link]

[17] Z. Lakache et al., “Phytochemical Screening and Antioxidant Properties of Methanolic Extracts and Different Fractions of Crataegus Azarolus Leaves and Flowers from Algeria,” International Food Research Journal, vol. 23, no. 4, pp. 1576-1583, 2016.

[Google Scholar] [Publisher Link]

[18] T.G. Gini, and G. Jeya Jothi, “Column Chromatography and HPLC Analysis of Phenolic Compounds in the Fractions of Salvinia Molesta Mitchell,” Egyptian Journal of Basic and Applied Sciences, vol. 5, no. 3, pp. 197-203, 2018.

[CrossRef] [Google Scholar] [Publisher Link]

[19] El Sayed Hassan Atwaa et al., “Antimicrobial Activity of Some Plant Extracts and Their Applications in Homemade Tomato Paste and Pasteurized Cow Milk as Natural Preservatives,” Fermentation, vol. 8, no. 9, pp. 1-16, 2022.

[CrossRef] [Google Scholar] [Publisher Link]

[20] Maria E. Garcia Denegri et al., “Neutralisation of the Pharmacological Activities of Bothrops Alternatus Venom by Anti-PLA₂ IgGs,” Toxicon, vol. 86, pp. 89–95, 2014.

[CrossRef] [Google Scholar] [Publisher Link]

[21] Sardar E. Gasanov, Ruben K. Dagda, and Eppie D. Rael, “Snake Venom Cytotoxins, Phospholipase A₂s, and Zn²⁺-Dependent Metalloproteinases: Mechanisms of Action and Pharmacological Relevance,” Journal of Clinical Toxicology, vol. 4, no. 1, 2014.

[CrossRef] [Google Scholar] [Publisher Link]

[22] Fatah Chérifi, Abdelkader Namane, and Fatima Laraba-Djebari, “Isolation, Functional Characterization and Proteomic Identification of CC2-PLA₂ from Cerastescerastes Venom: A Basic Platelet- Aggregation-Inhibiting Factor,” Protein Journal, vol. 33, pp. 66-77, 2014.

[CrossRef] [Google Scholar] [Publisher Link]

[23] Nwune Hope Chinyere, Mohammed Adamu Milala, and Hassan Zannah, “Effects of Aqueous Root Extract of Annona Senegalensis on Bitisarietans Venom Protease and Phospholipase A₂ Activities,” Journal Pharmaceutical Biomedical Science, vol. 6, no. 8, 2016.

[Google Scholar] [Publisher Link]

[24] Rafael Sutti et al., “Purification and Characterization of a Hyaluronidase from Venom of the Spider Vitalius Dubius (Araneae, Theraphosidae),” Journal of Venomous Animals and Toxins Including Tropical Diseases, vol. 20, pp. 1-7, 2014.

[CrossRef] [Google Scholar] [Publisher Link]

[25] U.K. Laemmli, “Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4,” Nature, vol. 227, pp. 680-685, 1970.

[CrossRef] [Google Scholar] [Publisher Link]

[26] B. Adzu et al., “Effect of Annona Senegalensis Rootbark Extracts on Naja Nigricotlis Nigricotlis Venom in Rats,” Journal of Ethnopharmacology, vol. 96, no. 3, pp. 507-513, 2005.

[CrossRef] [Google Scholar] [Publisher Link]

[27] Upasana Puzari, and Ashis K. Mukherjee, “Recent Developments in Diagnostic Tools and Bioanalytical Methods for Analysis of Snake Venom: A Critical Review,” Analytica Chimica Acta, vol. 1137, pp. 208−224, 2020.

[CrossRef] [Google Scholar] [Publisher Link]

[28] Makoto Murakami, Hiroyasu Sato, and Yoshitaka Taketomi, “Updating Phospholipase A₂ Biology,” Biomolecules, vol. 10, no. 10, pp. 1-32, 2020.

[CrossRef] [Google Scholar] [Publisher Link]

[29] M.S. Vineetha, Bhavya Janardhan, and S. Sunil, “Biochemical and Pharmacological Neutraliuzation of Indian Saw Scaled Viper Snake Venom by Canthium Parviflorum Extxracts,” Indian Journal of Biochemistry and Biophysics, vol. 54, no. 5, pp. 173-185, 2017.

[Google Scholar] [Publisher Link]

[30] Anant Deshwal et al., “A Meta-Analysis of the Protein Components in Rattlesnake Venom,” Toxins, vol. 13, no. 6, pp. 1-28, 2021.

[CrossRef] [Google Scholar] [Publisher Link]

[31] Zainab Umar Abdullahi et al., “Bactericidal Effects of Snake Venom Phospholipases A₂: A Systematic Review and Analysis of Minimum Inhibitory Concentration,” Physiologia, vol. 3, no. 1, pp. 30-42, 2023.

[CrossRef] [Google Scholar] [Publisher Link]

[32] Jeremy M. Berg, John L. Tymoczko, and Lubert Stryer, Biochemistry, 6^th ed., W. H. Freeman and Company, New York, 2007.

[33] Silvana L. Marunak et al., “Isolation of Biological Characterization of a Basic Phospholipase A₂ from Bothrop Jararacussu Snake Venom,” Biocell, Mendoza Journal, vol. 31, no. 3, pp. 3-8, 2007.

[Google Scholar]

[34] D.F.J. Ketelhut et al., “Isolation Characterization and Biological Activity of Acidic Phospholipase A₂ Isoforms from Bothrops Jararacussu Snake Venom,” Biochemistry, vol. 85, no. 10, pp. 983-991, 2003.

[CrossRef] [Google Scholar] [Publisher Link]