The medical herb Phyllanthus amarus play a crucial role in indigenous medicine. Therapeutically, these plants’ extract acts as potential players that inhibits several digestive enzymes that are relevant to the management of Peptic ulcers and Diabetes Mellitus, which occur due to the overproduction of such enzymes. Evaluation of inhibitory effect of this extract was carried out against Pepsin, α-amylase, Trypsin enzymes along with the effect of thermal stability and ammonium sulphate precipitation on these inhibitory assays. P. amarus leave’s extract with different concentration gradients were used in this research analysis. Results obtained along with the literature analysis revealed photochemical compounds such as polyphenols causes inhibitory nature in the extract. Maximal percentage of inhibition of amylase, pepsin and trypsin were found to be 71% (0.32 mg/ml), 85% (0.08 mg/ml) and 87% (1.28 mg/ml) respectively. In thermal stability assay the maximum percentage of inhibition for amylase, pepsin and trypsin was observed at 30% (80°C), 68% (4°C) and 5% (37°C). Enzymes inhibitory assays on ammonium sulphate precipitation elicited maximum percentage of inhibition for amylase, pepsin and trypsin as 42% (at 45% of (NH4)2SO4), 58% (at 15% of (NH4)2SO4) and 40% (at 30% of (NH4)2SO4) respectively. This research concluded that Phyllanthus amarus leave extracts are potential inhibitors of α-amylase, pepsin and trypsin enzymes. Ammonium sulphate precipitation was helpful to purify the polyphenols the active compounds to a good extend. Also, thermal stability was helpful to check the stability of these active photochemical compounds present in the extract. Thus, P. amarus is an effective inhibitor to be used as supplements in the disease management.
Amit, R. and Pushpa, P. (2016) Assessment of Mechanism of Action of Antidiabetic Activity of Calocybe indica by Enzyme Inhibitory Activity. Biosciences Biotechnology Research Asia, 13, 2117-2123. https://doi.org/10.13005/bbra/2372
[3]
Rai, V. and Kumar, P. (2012) Epidemiology Study of Glucose-6-Phosphate Dehydrogenase Deficiency in Scheduled Caste Population of India. Journal of Anthropology, 2012, Article ID: 984180. https://doi.org/10.1155/2012/984180
[4]
Nagral, A. (2012) Gaucher Disease. Journal of Clinical and Experimental Hepatology, 5, 113-121.
[5]
Mattar, R., Mazo, D.F.C. and Carrilho, J. (2012) Lactose Intolerance: Diagnosis, Genetic and Clinical Factors. Clinical and Experimental Gastroenterology, 5, 113-121. https://doi.org/10.2147/CEG.S32368
[6]
Abdel-Fattah, Y.R., Soliman, N.A., El-Toukhy, N.M., El-Gendi, H. and Ahmed, R.S. (2013) Production, Purification and Characterization of Thermostable α-Amylase Produced by Bascillus licheniformis Isolate AI20. Journal of Chemistry, 2013, Article ID: 673173. https://doi.org/10.1155/2013/673173
[7]
Afrapoli, F.M., Asghari, B., Saeidnia, S., Anjani, Y., Mirjani, M., Malmir, M., Bazaz, R.D., Hojakhoondi, A., Salehi, P., Hamburger, M. and Yassa, N. (2012) In Vitro α-Glucosidase Inhibitory Activity of Phenolic Constituents from Aerial Parts of Polygonum hyrcarnium. DARU Journal of Pharmaceutical Sciences, 20, Article No. 37. https://doi.org/10.1186/2008-2231-20-37
[8]
Charter, P.I., Wicox, M., Cherry, P., Herford, A., Mustar, S., Wheater, H., Brownlee, I., Seal, C. and Pearson, J. (2016) Inhibitory Activity of Extracts of Hebridean Brown Seaweeds on Lipase Activity. Journal of Applied Psychology, 28, 1303-1313. https://doi.org/10.1007/s10811-015-0619-0
[9]
Uddin, M.A., Uddin, M., Lira, D.N. and Rouf, A.S.S. (2014) Assessment of Cytotoxic Activities of Phyllanthus amarus and Monestera deliciosa. Journal of Applied Pharmaceutical Science, 4, 110-113.
[10]
Bongu, S.B.R., Sagree, S., Bellamkonda, R. and Desireddy, S. (2016) Protective Role of Aqueous Extract of Phyllanthus amarus on Oxidative Stress in Pancreas of Streptozotocin Induced Diabetic Male Wistar Rats. Journal of Experimental and Applied Animal Sciences, 2, 23-30. https://doi.org/10.20454/jeaas.2016.1087
[11]
Umoh, E.D., Akpabio, U.D. and Udo, I.E. (2013) Photochemical Screening and Nutrient Analysis of Phyllanthus amarus. Asian Journal of Plant Sciences and Research, 3, 116-122.
[12]
Tjandrawinata, R.R., Susanto, L.W. and Nofiarny, D. (2017) The Use of Phyllanthus niruri L. as an Immune Modulator for the Treatment of Infectious Disease in Clinical Setting. Asian Pacific Journal of Tropical Disease, 7, 132-140. https://doi.org/10.12980/apjtd.7.2017D6-287
[13]
Paramayani, I., Paraton, H. and Maát, S. (2005) Comparison of Success Rate of Vaginal Candidiasis Treatment between Ketoconazole and Combination of Ketoconazole-Phyllanthus niruri Extract. Dexa Media, 18, 97-102.
[14]
Micali, S., Sighinolfi, M.C., Celia, A., Destefani, S., Grande, M., Cicero, A.F. and Bianchi, G. (2006) Can Phyllanthus niruri Effect the Efficiency of Extracorporeal Shock Wave Lithotripsy for Renal Stones? A Randomized Prospective Long Term Study. The Journal of Urology, 176, 1020-1022. https://doi.org/10.1016/j.juro.2006.04.010
[15]
Yang, Y., Xiu, J., Liu, J., Zhang, L., Li, X., Xu, Y., Quin, C. and Zhang, L. (2013) Chebulagic Acid, a Hydrolyzable Tannin, Exhibited Antiviral Activity in Vitro and in Vivo against Human Enterovirus 71. International Journal of Molecular Sciences, 14, 9618-9627. https://doi.org/10.3390/ijms14059618
[16]
Dirjomulijono, M. and Tjandrawinta, R.R. (2008) Clinical Trials Involving Phyllanthus Species. Journal of Molecular Sciences, 1, 183-313.
[17]
Kazeem, M.I., Adamson, J.O. and Ogunwande, I.A. (2013) Modes of Inhibition of α-Amylase and α-Glucosidase by Aqueous Extract of Morinda lucida Benth Leaf. BioMed Research International, 2013, Article ID: 527570. https://doi.org/10.1155/2013/527570
[18]
Sarasetyaningtyas, P.V., Hadinegoro, S.R. and Munasir, Z. (2006) Randomized Controlled Trial of Phyllanthus niruri Linn Extract. Paediatrica Indonesia, 2, 152-159.
[19]
Tamil, I.G., Dineshkumar, B., Nandhakumar, M., Senthilkumar, M. and Mitra, A. (2010) In-Vitro Study on α-Amylase Inhibitory Activity of an Indian Medical Plant Phyllanthus amarus. Indian Journal of Pharmacology, 42, 280-282.
[20]
Mcdougall, G.J. and Stewart, D. (2005) The Inhibitory Effects of Berry Polyphenols on Digestive Enzymes. BioFactors, 23, 189-195. https://doi.org/10.1002/biof.5520230403
[21]
Glenn, M.J.V., Glenn, M. and Gomer, R.H. (2013) Trypsin Potentiates Human Fibrocyte Differentiation. PLoS ONE, 8, e70795. https://doi.org/10.1371/journal.pone.0070795
[22]
Kumwenda, C., Dewey, K.G., Hemsworth, J., Ashorn, P., Maleta, K. and Haskell, M.J. (2013) Lipid-Based Nutrient Supplements Do Not Decrease Breast Milk Intake of Malawian Infants. American Journal of Clinical Nutrient, 99, 617-623. https://doi.org/10.3945/ajcn.113.076588
[23]
Wingfield, P.T. (2001) Protein Precipitation Using Ammonium Sulfate. Current Protocol in Protein Science, 2001, 1-10.
[24]
Singh, K.P., Kumar, A. and Prasad, R. (2013) Pepsin Assay of the Easiest Approach for Prescreening of HIV Protease Inhibitors. Journal of Pharmaceutical and Scientific Innovation, 2, 53-56.
[25]
Subramanian, R., Asmawi, M.Z. and Sadikun, A. (2008) In Vitro Alpha-Glucosidase Enzyme Inhibitory Effects of Andrographis paniculata Extract and Andrographolide. Acta Biochimica Polonica, 55, 391-398. https://doi.org/10.18388/abp.2008_3087
[26]
Boisen, S. and Djurtoft, R. (1981) Trypsin Inhibitor from Wheat Endosperm Purification and Characterization. Cereal Chemistry, 58, 460-463.
[27]
Manasa, N. and Ashadevi, J.S. (2015) Impact of Phyllanthus amarus Extracts on Antioxidant Enzymes in Drosophila Melanogaster. Journal of Applied Biology & Biotechnology, 3, 43-47.
[28]
Verma, S., Sharma, H. and Garg, M. (2014) Phyllanthus amarus. Journal of Pharmacology and Photochemistry, 1, 224-236.
[29]
Lawson-Evi, P., Ekul-Gadegbeku, K., Agbonan, A., Akilkokou, K., Creppy, E. and Gbeassor, M. (2011) Antidiabetic Activity of Phyllanthus amarus Schum and Thonn (Euphorbiaceae) on Alloxan Induced Diabetes in Male Wistar Rats. Journal of Applied Sciences, 11, 2968-2973. https://doi.org/10.3923/jas.2011.2968.2973
[30]
Oyewole, O., Taiwo, A.O. and Quadri, N. (2015) Evaluation of Hypoglycemic Efficacy of Methanolic Extracts of Moringa oleifera and Phyllanthus amarus in Diabetic Rats. British Biotechnology Journal, 5, 98-102. https://doi.org/10.9734/BBJ/2015/13164
[31]
Abdulla, A., Peeters, A., Courten, M. and Stoelwinder, J. (2010) The Magnitude of Association between Overweight and Obesity and the Risk of Diabetes: A Meta-Analysis of Prospective Cohort Studies. Diabetes Research and Clinical Practice, 89, 309-319. https://doi.org/10.1016/j.diabres.2010.04.012
[32]
Majeed, W., Khaliq, T., Aslam, B., Khan, J.A. and Iftikhar, A. (2015) Medicinal Plants with Gastroprotective Potential. Bangladesh Journal of Pharmacology, 10, 588-603.
[33]
Yuan, L., Wang, M., Zhang, X. and Wang, Z. (2017) Effects of Protease and Non-Starch Polysaccharide Enzyme on Performance, Digestive Function, Activity and Gene Expression of Endogenous Enzyme of Broilers. PLoS ONE, 12, e0173941. https://doi.org/10.1371/journal.pone.0173941
[34]
Buchner, N., Krumbein, A., Rohn, S. and Kroh, L. (2006) Effect of Thermal Processing on the Flavonols Rutin and Quercetin. Rapid Communications in Mass Spectrometry, 20, 3229-3235. https://doi.org/10.1002/rcm.2720
[35]
Nuruddin, M.N. and Al-Jasabi, S. (2016) Characterization of Polyphenol Oxidase from Phyllanthus amarus Leaves. The International Journal of Medical Science, 1, 29-32.
[36]
Purwanto, M.G.M. (2015) The Role and Efficiency of Ammonium Sulphate Precipitation in Purification Process of Papain Crude Extract. Procedia Chemistry, 18, 127-131. https://doi.org/10.1016/j.proche.2016.01.020
[37]
Zheng, Z.Z., Chen, L.H., Liu, S.S., Deng, Y., Zheng, G.H., Gu, Y. and Ming, Y.L. (2016) Bioguided Fraction and Isolation of the Antitumor Components from Phyllanthus niruri L.. Biomedical Research International, 2016, Article ID: 9729275. https://doi.org/10.1155/2016/9729275
[38]
Shetti, A.A., Sanakal, R.D. and Kaliwal, B.B. (2012) Antidiabetic Effect of Ethanolic Leaf Extract of Phyllanthus amarus in Alloxan Induced Diabetic Mice. Asian Journal of Plant Science and Research, 2, 11-15.
