In Vitro Screening of Seed Extracts of Medicinal Plants for Protease Inhibitory Activity

  • Faiyaz Khudaboddin Shaikh (1) Department of Biochemistry, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad, Maharashtra, India; (2) Department of Biotechnology, MGM’s Institute of Biosciences and Technology, Aurangabad, Maharashtra, India
  • Sarwan W. Bradosty Department of Biology, Cihan University-Erbil, Kurdistan Region, Iraq
  • Saber W. Hamad Department of Field Crops Production, College of Agriculture, Salahaddin University - Erbil, Erbil, Kurdistan Region, Iraq
  • Ashok A. Shinde Department of Biotechnology, MGM’s Institute of Biosciences and Technology
Keywords: Dot blot, protease inhibitors, Proteases, Protein, Trypsin


Protease inhibitors (PIs) are deployed in the plant kingdom as storage proteins or peptides, regulators of endogenous proteases, and plant protection agents against insect pests and pathogen attack. In humans, they are identified as chemopreventive agents against a range of cancers and have potential as drug to treat an array of disease associated with aberrant activity of proteases. The present investigation reports PIs activity data from 30 medicinal plants. The screening for PIs activity was done by dot blot assay using X-ray film coated with gelatin. Among screened seed extracts, Albizia lebbeck, Raphanus sativus, Mucuna pruriens, Achyranthes aspera, and Coffea arabica showed high inhibitory activities with trypsin protease. Most of seed extracts exhibited moderate activity, whereas Ocimum sanctum showed moderate to low activity against trypsin. The presence of varied protein content is reported from all seed extracts with highest in A. lebbeck (50.0 ± 3.4 mg/ml). The data produced in the present investigation could be helpful for further exploration of PIs as therapeutic agent.


Download data is not yet available.


A. J. Barrett, N. D. Rawlings and J. J. F. Woessner. “Hand Book of Proteolytic Enzymes”. London: Acadamy Press, 1998.

S. G. Fan and G. J. Wu. “Characteristics of plant proteinase inhibitors and their applications in combating phytophagous insects”. Botanical bulletin of Academia Sinica, Vol. 46, pp. 273-292, 2005.

A. N. Flynn and A. G. Buret. “Proteinase-activated receptor 1 (PAR-1) and cell apoptosis”. Apoptosis, Vo. 9, pp. 729-737, 2004.

G. A. Joanitti, S. M. Freitas and L. P. Silva. “Proteinaceous protease inhibitors: Structural features and multiple functional faces”. Current Enzyme Inhibition, Vol. 2, no. 3, pp. 199-217, 2006.

P. K. Lawrence and K. R. Koundal. “Plant protease inhibitors in control of polyphagous insects”. Electronic Journal of Biotechnology. Vol. 1, pp. 93-109, 2002.

D. Ivanov, C. Emonet, F. Foata, M. Affolter, M. Delly, M. Fisseha, S. Blum-Sperisen, S Kochhar and F. Arigoni. “A serpin from the gut bacterium Bifidobacterium longum inhibits eukaryotic elastase like serine proteases”. Journal of Biological Chemistry, vol. 281, no. 25, pp. 17246-17252, 2006.

A. J. Barrett, N. D. Rawlings and E. A. O’Brien. “The MEROPS database as a protease information system”. Journal of Structural Biology, vol. 134, pp. 95-102, 2001.

X. S. Puente, L. M. Sanchez, C. M. Overall and C. LopezOtin. “Human and mouse proteases: A comparative genomic approach.” Nature Reviews Genetics, vol. 4, pp. 544-558, 2003.

D. N. Rawlings, P. D. Tol1e and A. J. Barrett. “Evolutionary families of peptidase inhibitors”. Biochemical Journal, vol. 378, pp. 705-716, 2004.

J. D. Tyndall, T. Nall and D. P. Fairlie. “Proteases universally recognize beta strands in their active sites.” Chemical Reviews, vol. 105, pp. 973-999, 2005.

A. J. Molehin, G. N. and D. P. Gobert. “McManus: Serine protease inhibitors of parasitic helminthes.” Parasitology, vol. 139, no. 6, pp. 681-695, 2012.

M. L. J. Korsinczky, H. J. Schirra and D. J. Craik. “Sunflower trypsin inhibitor-1”. Current Protein and Peptide Science, vol. 5, p. 351, 2004.

M. H. Lingaraju and L. R. Gowda. “A Kunitz trypsin inhibitor of Entada scandens seeds: Another member with single disulfide bridge.” Biochimica et Biophysica Acta, vol. 1784, pp. 850-855, 2008.

P. R. Shewry. “Tuber storage proteins”. Annals of Botany, vol. 91, pp. 755-769, 2003.

N. D. Rawlings, A. J, Barrett and A. Bateman. “MEROPS: The database of proteolytic enzymes, their substrates and inhibitors”. Nucleic Acids Research, vol. 40, pp. 343-350, 2012.

C. Fermi and L. Pernossi. “Untersuchungen uber die enzyme, ver-gleichende studie”. Zeitschrift für Hygiene und Infektionskrankheiten, vol. 18, pp. 83-89, 1894.

H. Fritz. Foreword. In: K. von der Helm, B. D. Korant and J. C. Cheronis, editors. “Proteases as Targets for Therapy”. Berlin: Springer-Verlag, pp. 5-6, 2000.

F. De Leo, M. Volpicella, F. Licciulli, S. Liuni, R. Gallerani and L. R. Ceci. “PLANT-PIs: A database for plant protease inhibitors and their genes”. Nucleic Acids Research, vol. 30, no. 1, pp. 347-348, 2002.

G. Fear, S. Komarnytsky and I. Raskin. “Protease inhibitors and their peptidomimetic derivatives as potential drugs.” Pharmacology and Therapeutics, vol. 113, pp. 354-368, 2007.

G. C. Mello, M. L. V. Oliva, J. T. Sumikawa, O. L. T. Machado, S. Marangoni, J. C. Novello and M. L. R. Macedo. “Purification and characterization of a new trypsin inhibitor from Dimorphandra mollis seeds”. Journal of Protein Chemistry, vol. 20, pp. 625-632, 2001.

S. Haq, S. Atif and R. Khan. “Protein proteinase inhibitor genes in combat against insects, pests, and pathogens: Natural and engineered phytoprotection”. Archives of Biochemistry and Biophysics, vol. 431, pp. 145-159, 2004.

J. W. Read and L. W. Haas. “Studies on the baking quality of flour as affected by certain enzyme actions. V. Further studies concerning potassium bromate and enzyme activity”. Cereal Chemistry, vol. 45, pp. 59-68, 1938.

M. V. Padul, R. D. Tak and M. S. Kachole. “Protease inhibitor (PI) mediated defense in leaves and flowers of pigeonpea (protease inhibitor mediated defense in pigeonpea)”. Plant Physiology and Biochemistry, vol. 52, pp. 77-82, 2012.

O. H. Lowry, N. J. Rosebrough, A. L. Farr and R. J. Randall. “Protein measurement with folin phenol reagent”. Journal of Biological Chemistry, vol. 193, pp. 265-275, 1951.

C. A. Ryan. “Protease inhibitors in plants: Genes for improving defenses against insects and pathogens”. Annual Review of Phytopathology, vol. 28, pp. 425-449, 1990.

P. J. Magee, R. Owusu-Apenten, M. J. McCann, C. I. Gill and I. R. Rowland. “Chickpea (Cicer arietinum) and other plantderived protease inhibitor concentrates inhibit breast and prostate cancer cell proliferation in vitro”. Nutrition and Cancer, vol. 64, pp. 741-748, 2012.

J. I. Sprent. “Legume Nodulation: A Global Perspective”. Ames, Iowa: Wiley-Blackwell, pp. 12, 2009.

N. Vergnolle. “Protease inhibition as new therapeutic strategy for GI diseases”. Gut, vol. 65 pp. 1215-1224, 2016.

C. Fields, P. Mallee, J. Muzard and G. U. Lee. “Isolation of Bowman-Birk inhibitor from soybean extracts using novel peptide probes and high gradient magnetic separation”. Food Chemistry, vol. 134, pp. 1831-1838, 2012.

How to Cite
Shaikh F, Bradosty S, Hamad S, Shinde A. In Vitro Screening of Seed Extracts of Medicinal Plants for Protease Inhibitory Activity. cuesj [Internet]. 30Jun.2019 [cited 16Apr.2024];3(1):61-5. Available from:
Research Article