Copyright (c) 2014 AJC
This work is licensed under a Creative Commons Attribution 4.0 International License.
Extraction and Purification of Lectin by Employing Anionic Surfactant Aerosol-OT Reversed Micellar System and Evaluation of its Antibacterial and Antioxidant Potentials
Corresponding Author(s) : Senthilkumar Rathnasamy
Asian Journal of Chemistry,
Vol. 26 No. 12 (2014): Vol 26 Issue 12
Abstract
Aloe vera is a high potential medicinal plant that contains many active biomolecules like proteins, sugars, amino acids and enzymes. In the present study, lectin from Aloe vera gel and Aloe vera leaf were extracted using non organic method. Sudden denaturation and incompatibleness in the conventional organic extraction method was overcome by biological extraction method. Lectin fractionation was done by ammonium sulphate precipitation and compared with reverse micellar extraction method. The protein content in the crude extracts was estimated to be 2.312 mg/mL and sample obtained after ammonium sulphate precipitation and reverse micellar extraction was found to be 1.77 and 2.068 mg/mL, respectively. The maximum activity fraction was collected and purified using cationic exchange chromatography on CM sepharose column. Purified fractions from the chromatographic column were collected and confirmed for the presence of lectin by native page analysis. The resulting fractions were assayed for haemagglutination activity using human erythrocytes, in vitro antibacterial activity using agar well diffusion method and in vitro antioxidant activity using reducing power assay. Results revealed that the extraction of lectin from Aloe vera leaf extracts using reverse micellar extraction is an effective method as compared to ammonium sulphate precipitation method and lectin was also found to possess high antioxidant activity with respective to the concentration of lectin and wide spectrum of antibacterial effect. A zone of inhibition against E. coli is (11 mm), of S. typhi (9 mm) is P. aeroginosa (9 mm), E. aerogenis (9 mm) and K. pneumonia (5 mm) were observed.
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References
T. Reynolds and A.C. Dweck, J. Ethnopharmacol., 68, 3 (1999); doi:10.1016/S0378-8741(99)00085-9.
M.S. Moghaddasi and S.K. Verma, Int. J. Biol. Med. Res., 2, 466 (2011).
A. Surjushe, R. Vasani and D.G. Saple, Indian J. Dermatol., 53, 163 (2008); doi:10.4103/0019-5154.44785.
S. Singh, P.K. Sharma, N. Kumar and R. Dudhe, Int. J. Pharm. Sci., 2, 1224 (2010).
M.D. Boudreau and F.A. Beland, J. Environ. Sci. Health C, 24, 103 (2006); doi:10.1080/10590500600614303.
M. Bhattacharya, S. Malik and A. Singh, J. Pharm. Res., 4, 4507 (2011).
W.D. Winters, Phytother. Res., 7, S23 (1993); doi:10.1002/ptr.2650070709.
Y.F. Hou, Y.B. Hou, Y.Y. Liu, Q. Guang and J.C. Li, J. Biomed. Biotechnol., Article ID 217342 (2010); doi:10.1155/2010/217342.
J. Shi, S.J. Xue, Y. Kakuda, S. Ilic and D. Kim, Process Biochem., 42, 1436 (2007); doi:10.1016/j.procbio.2007.07.015.
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C. Bies, C.-M. Lehr and J.F. Woodley, Adv. Drug Deliv. Rev., 56, 425 (2004); doi:10.1016/j.addr.2003.10.030.
J. Li, H. Wu, J. Hong, X. Xu, H. Yang, B. Wu, Y. Wang, J. Zhu, R. Lai, X. Jiang, D. Lin, M.C. Prescott and H.H. Rees, PLoS ONE, 3, e2381 (2008); doi:10.1371/journal.pone.0002381.
L.G. Barrientos and A.M. Gronenborn, Mini Rev. Med. Chem., 5, 21 (2005); doi:10.2174/1389557053402783.
M. Kaur, J. Singh and S.S. Kamboj, J. Pharm. Res., 4, 2441 (2011).
A.B. Hemavathi, H.U. Hebbar and K.S.M.S. Raghavarao, Sep. Purif. Technol., 71, 263 (2010); doi:10.1016/j.seppur.2009.11.026.
C.O. Nascimento, R.M.P.B. Costa, R.M.S. Araújo, M.E.C. Chaves, L.C.B.B. Coelho, P.M.G. Paiva, J.A. Teixeira, M.T.S. Correia and M.G. Carneiro-da-Cunha, Process Biochem., 43, 779 (2008); doi:10.1016/j.procbio.2008.02.021.
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A.B. Hemavathi, U.H. Hebbar and K.S.M.S. Raghavarao, J. Chem. Technol. Biotechnol., 82, 985 (2007); doi:10.1002/jctb.1769.
C. Jolivalt, M. Minier and P. Renon, Biotechnol. Prog., 9, 456 (1993); doi:10.1021/bp00023a003.
M. Dekker, R. Hilhorst and C. Laane, Anal. Biochem.., 178, 217 (1989); doi:10.1016/0003-2697(89)90628-3.
K.E. Cöklen and T.A. Hatton, Biotechnol. Prog., 1, 69 (1985); doi:10.1002/btpr.5420010113.
M.J. Pires, M.R. Aires-Barros and J.M.S. Cabral, Biotechnol. Prog., 12, 290 (1996); doi:10.1021/bp950050l.
Y.- Yu, Y. Chu and J.-Y. Ji, J. Colloid Interf. Sci., 267, 60 (2003); doi:10.1016/S0021-9797(03)00450-8.
J.R.S. Alves, L.P. Fonseca, M.T. Ramalho and J.M.S. Cabral, Biochem. Eng. J., 15, 81 (2003); doi:10.1016/S1369-703X(02)00181-X.
M.M. Bradford, Anal. Biochem., 72, 248 (1976); doi:10.1016/0003-2697(76)90527-3.
B.D. Hames and D. Rickwood, Gel Electrophoresis of Proteins-A Practical Approach, Oxford University Press, New York, edn 2 (1990).
U.K. Laemmli and M. Favre, J. Mol. Biol., 80, 575 (1973); doi:10.1016/0022-2836(73)90198-8.
I.E. Liener and E.G. Hill, J. Nutr., 49, 609 (1953).