Journal of Environmental Biology

Extraction of bioactive compounds from Limonia acidissima to explore their therapeutic potential

P. Kale¹, M. Dake¹*, J. Manure² and A. Tamboli³     

¹Department of Biotechnology, Protein Biochemistry Research Center, Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Pune-411 033, India

²Department of Pharmaceutical Quality Assurance, Appasaheb Birnale College of Pharmacy, Sangali-416 416, India

³Department of Pharmaceutical Chemistry, Sahyadri College of Pharmacy, Solapur-413 307, India 

Received: 24 October 2025                   Revised: 19 February 2026                   Accepted: 28 February 2026

*Corresponding Author Email : manjusha.dake@dpu.edu.in                     *ORCiD: https://orcid.org/0000-0001-5578-4939

Abstract

Aim: This study evaluates the bioactivity and molecular mechanisms of Limonia acidissima (wood apple) leaf extracts across multiple therapeutic targets.

Methodology: Solvent extracts of L. acidissima (ethanol 70%, ethyl acetate 70%, and methanol 70%) were assessed for bioactivity screening, antimicrobial profiling, phytochemical characterization, and molecular docking analysis.

Results: Ethyl acetate extract exhibited higher protease inhibition (67.45±1.88%, p<0.001) and broad-spectrum antimicrobial activity. Ethanol extract showed high anti-diabetic potential (75.22±1.26% α-amylase inhibition, statistically comparable to acarbose, p>0.05) with significant dose-dependent anticancer activity against MCF-7 breast cancer cells (linear slope = 0.311% μl^-1, R² = 0.976, p < 0.001). Methanol extract showed the highest antioxidant activity (78.15±0.99%, higher than ascorbic acid control by 16%, p<0.01). Flash chromatography identified seven chemical entities, of which three—bergapten, methyl palmitate, and oleanolic acid—were selected for molecular docking analysis. Molecular docking revealed oleanolic acid as a promising compound with strong binding affinities across therapeutic targets: trypsin (-8.61 kcal mol^-1), estrogen receptor (-6.80 kcal mol^-1), and insulin receptor (-6.99 kcal mol^-1). Methyl palmitate showed the strongest COX-2 affinity (-8.07 kcal mol^-1) with 15 distinct interaction points.

Interpretation: The findings suggest the reactivity and stability of extracted bioactive compounds as a protease inhibitor and their potential to develop effective insecticidal agents as well as therapeutic candidate. Exploring the role of protease inhibitors as a biocontrol agent can be an effective way to replace hazardous chemical pesticides producing harmful effect on the environment.

Key words: Ethnopharmacology, Limonia acidissima, Molecular docking, Multi-target therapeutics, Protease inhibitors

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