Date of Award
Summer 8-2026
Document Type
Thesis
Degree Name
Master of Science (MS)
Department
Pharmaceutical Sciences
First Advisor
Aftab Ahmed
Second Advisor
Kamaljit Kaur
Third Advisor
Innokentiy Maslennikov
Abstract
Moringa oleifera (drumstick tree) is widely recognized in Traditional and Complementary Medicine (T&CM) for its extensive therapeutic properties; however, its seed proteome remains underrepresented in public sequence repositories, limiting our understanding of its molecular mechanisms. This study evaluated the M. oleifera seed proteome using an integrated top-down and bottom-up proteomic workflow paired with tandem mass spectrometry (LC-MS/MS). Crude seed proteins were extracted, dialyzed, and fractionated using size exclusion chromatography (SEC) and heparin affinity chromatography. To optimize enzymatic cleavage and peptide recovery, proteins were subjected to performic acid oxidation followed by digestion using enzymes trypsin or pepsin (both in-solution and immobilized). MS/MS spectra were processed using PEAKS Studio X for de novo sequencing and homologous database searches against NCBI Moringa and SwissProt Viridiplantae repositories. Tryptic digestion of the crude matrix and SEC fractions yielded 728 high-confidence peptides (FDR 0.8%), successfully profiling 82 distinct protein groups within peak fractions. Heparin affinity purification isolated major functional clusters, including 2S albumin precursors, chitin-binding proteins (Mo-CBP2, Mo-CBP4, CBP3), flocculating proteins, and small cysteine-rich antimicrobial peptides (morintides mO1-mO6). Gene Ontology (GO) functional analysis categorized the identified proteome primarily into metabolite interconversion enzymes (39.5%), regulatory proteins (17.1%), and cytoskeletal proteins (16.4%) by protein class. While binding (39.5%) and catalytic activity (29.7%) dominated molecular function. These findings bridge the gap between traditional ethnomedicinal applications and modern molecular pharmacology by expanding the known seed proteome of M. oleifera and identifying novel functional peptides for future therapeutic exploration.
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.
Recommended Citation
Banatin, PBP. Evaluation of Drumstick Tree (Moringa oleifera) Seeds Proteome by Tandem Mass Spectrometry. [master’s thesis or dissertation]. Irvine, CA: Chapman University; Year. https://doi.org/10.36837/chapman.000759