Isolation, Characterizations, Anti-Cancer, Cytotoxicity, in Silico Toxicity, and Antimicrobial Evaluations of Moringa Oleifera Seed Protein

Abstract

Background: Moringa oleifera seed, which is widely utilized as an affordable source of domestic water treatment, is also reputable for its various pharmacological applications globally. Aim: This study evaluates the toxicity and pharmacological potentials of Moringa oleifera seed protein while establishing the chemical profile. Method: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and liquid chromatography coupled with electrospray ionization mass spectrometry (LC-ESI-MS/MS) were used to analyze the protein/peptide content of the purified protein. The protein was also investigated for in vitro anticancer potential on Hela and MDA-MB-231 cell lines, cytotoxic on 3T3 cell line, and toxicity to brine shrimps. In addition, the antimicrobial action of the protein (polypeptide) was evaluated. Results and Discussion: The MALDI-MS revealed three protein moieties with values of 3.4, 4.6, and 6.9 kDa. On the other hand, LC-ESIMS/MS analysis revealed dipeptides and tripeptides, which include serinylarginine, leucylproline, leucylmethionyl-glycine, isoleucyl-glycyl-methinine, glycyl-arginyl-aspartic acid, isoleucyl-glutamyl-methionine, diphenylalanine, asparaginyl-aspartyl-histidine, seryl-tyrosyl-tyrosine, phenylalanyl-asparaginyl-tyrosine, and propanolyl-trytophanyl-glycine. The seed protein exhibited extremely low toxicity on both 3T3 and MDA-MB-231 breast cancer cell lines, while the standards, doxorubicin, and cycloheximide had IC50 = 0.5 ± 0.07 and 0.8 ± 0.10 μg/mL, respectively. Further, the protein showed no antibacterial or antifungal activity against all tested organisms. The protein also exhibited no lethality against brine shrimp. The in-silico toxicity results indicated that the peptides are not immunotoxic, carcinogenic, or mutagenic as most belonged to the tox class V except serylarginine predicted for tox class IV (harmful if swallowed). Hence, the intake of the protein remnant in water could be within the tolerable limit. Conclusion: The results obtained suggested that the protein content of the Moringa oleifera seed is non-cytotoxic. The result further validates the safety potential of the defatted and debittered seed material used as potential food sources for both humans and animals.

Role of miRNA in pathogenesis, diagnosis, and prognosis in hepatocellular carcinoma

“…Many genetic and epigenetic factors like methylation are involved in the process (Fawzi et al, 2020). To treat the disorder, various attempts are being made that involves use of natural substances (Olubunmi et al, 2021), modification of immune system (Mukherjee et al, 2022) and others. With over 700,000 deaths annually worldwide, hepatic cancer is responsible for the second‐highest number of deaths in cancer patients (Wan & El‐Nezami, 2018).…”