PURIFICATION AND ANTIOXIDANT ACTIVITY OF ALOE VERA LEAF LECTIN

Year 2012, Volume: 1 Issue: 42(1), 1 - 11, 10.01.2014

N.* Ozsoy E.* Candoken N.* Akev N. Ozsoy, E. Candoken, N. Akev* N. Ozsoy, E. Candoken, N. Akev*

Abstract

Aloe vera (L.) Burm. fil. lektinin saflaştırılması için, yeni ve hızlı bir
kromatografi yöntemi olan, ovalbumin bağlı-siyanojen bromür (CNBr)
ile aktive edilmiş Sefaroz 4B, afinite kromatografisi tanıtılmaktadır.
Lektin, kromatografi sonucunda 60 kez saflaştırıldı. Poliakrilamid disk jel
elektroforezi (PAGE) ile homojen olduğu gözlemlenen lektinin molekül
ağırlığı bu yöntemle, 45 000 kD olarak saptandı. Sodyum dodesil sülfat
(SDS)-PAGE ile altbirim molekül ağırlığı 14 400 kDa olarak belirlenen
lektinin, üç alt birimden oluştuğu ve bunun da daha önce tarafımızdan
kısmen saflaştırılmış ve karakterize edilmiş olan Aloctin I ile uyum içinde
olduğu sonucuna varıldı. Saflaştırılan lektin, DPPH·
radikal süpürme tayini
ile değerlendirildiğinde antioksidan etki göstermedi.

Keywords

Aloe vera, lectin, purification, affinity chromatography, antioxidant activity

PURIFICATION AND ANTIOXIDANT ACTIVITY OF ALOE VERA LEAF LECTIN

Abstract

A new and rapid affinity chromatography method based on cyanogen
bromide (CNBr)-activated Sepharose 4B bound-ovalbumin is presented for
the purification of the main lectin present in Aloe vera (L.) Burm. fil. The
lectin was purified 60 fold to apparent homogeneity in native polyacrylamide
disc gel electrophoresis (PAGE) showing an apparent molecular weight of
45000 kDa. The fact that sodium dodecyl sulfate (SDS)-PAGE gave a subunit
molecular weight of 14 400 kDa tends to propose that the lectin is composed
of three subunits and thus is in agreement with Aloctin I previously partially
purified and characterized by us. The lectin did not exhibit antioxidant effect
as assessed by the DPPH·
radical-scavenging assay.

Keywords

Aloe vera, lectin, purification, affinity chromatography, antioxidant activity

References

• 1. Capasso, F., Borrelli, F., Capasso, R., Di Carlo, G., Izzo, A.A., Pinto, L., Mascolo, N., Castaldo, S., Longo, R., Aloe and its therapeutic use, Phytother Res., 12, S124-S127 (1998).
• 2. Boudreau, M.D., Beland, F.A., An evaluation of the biological and toxicological properties of Aloe barbadensis Miller, Aloe vera. J Environ Science Health, 24, 103-154 (2006).
• 3. Reynolds, T., Aloes, the Genus Aloe, CRC Press LLC. USA (2004). 4. Park, Y.I., Lee, S.K., New Perspectives on Aloe. Springer Science + Business Media, LLC. US (2006).
• 5. Choi, S., Chung, M.H., A review on the relationship between Aloe vera components and their biologic effects. Seminars in Integrative Medicine, 1, 53-62 (2003).
• 6. Eshun, K., He, Q., Aloe vera: A valuable ingredient for the food, pharmaceutical and cosmetic industries-a review. Crit Rev Food Sci Nutr, 44, 91-96 (2004).
• 7. Saada, H.N., Ussama, Z.S., Mahdy, A.M., Effectiveness of Aloe vera on the antioxidant status of different tissues in irradiated rats. Pharmazie, 58, 929-931 (2003).
• 8. Saito, H., Purification of active substances of Aloe arborescens Miller and their biological and pharmacolgical activity. Phytother Res, 7, S14-S19 (1993).
• 9. Koike, T., Beppu, H., Kuzuya, H., Maruta, K., Shimpo, K., Suzuki, M., Titani, K., Fujita, K., A35 kDA mannose-binding lectin with haemagglutinating and mitogenic activities from “Kidachi Aloe” Aloe arborescens Miller. var. natalensis Berger, J Biochem, 118, 1205-1210
• 10. Yagi, A., Machii, K., Nishimura, H., Shida, T., Nishioka, I., Effect of aloe lectin on deoxyribonucleic acid synthesis in baby hamster kidney cells. Experientia, 41, 669-671 (1985).
• 11. Imanishi, K., Aloctin A, an active substance of Aloe arborescens Miller as an immunomodulator, Phytother Res, 7, S20-S22 (1993).
• 12. Sharon, N., Lis, H., Lectins: cell-agglutinating and sugar-specific proteins, Science, 177, 949-958 (1972).
• 13. Lis, H., Sharon, N., Affinity chromatography for the purification of lectins (a review), J. Chromatogr, 215, 361-372 (1981).
• 14. Suzuki, I., Saito, H., Inoue, S., Migita, S., Takahachi, T., Purification and characterization of two lectins from Aloe arborescens Mill, J Biochem, 85, 163-171 (1979).
• 15. Chung, Y.J., Park, Y.I., Purification and partial characterization of lectin from Aloe vera, J. Inst. Biotechnol., Korea University, 8, 71-78 (1996).
• 16. Akev, N., Can, A., Separation and some properties of Aloe vera L. leaf pulp lectins. Phytother, Res., 13, 489-493 (1999).
• 17. Lis, H., Sharon, N., Soybean (Glycine max) agglutinin, Methods Enzymol, 28 Part B, 366-368 (1972).
• 18. Warburg, O., Christian, W., Isoluerung und Kristallisation des Gärungsferments Enolase, Biochem, Z., 310, 384-421 (1941).
• 19. Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J., Protein measurement with the Folin phenol reagent, J. Biol. Chem., 193, 265- 275 (1951).
• 20. Brand-Williams, W., Cuvelier, M. E., Berset, C., Use of a free radical method to evaluate antioxidant activity. Lebensmittel-Wissenchaft und Technologie (Food Science and Technology), 28, 25-30 (1995).
• 21. De Mejía, E.G., Prisecaru, V.I., Lectins as bioactive plant proteins: a potential in cancer treatment, Crit. Rev. Food Sci. Nutr., 45, 425-445 (2005).
• 22. Akev, N., Malkoç, S., Can, A., Further characterization of Aloe vera leaf pulp lectins, J. Fac. Pharm. Istanbul., 35-36, 1-11 (2002-2003).
• 23. Kardošová, A., Machová, E., Antioxidant activity of medicinal plant polysaccharides, Fitoterapia, 77, 367-373 (2006).
• 24. Wu, J.H., Xu, C., Shan, C.Y., Tan, R.X., Antioxidant properties and PC12 cell protective effects of APS-1, a polysaccharide from Aloe vera var. Chinensis, Life Sci., 78, 622-630 (2006).
• 25. Chun-hui, L., Chang-hai, W., Zhi-liang, X., Yi, W., Isolation, chemical characterization and antioxidant activities of two polysaccharides from the gel and the skin of Aloe barbadensis Miller irrigated with sea water, Process Biochemistry, 42, 961-970 (2007).
• 26. Hu, Y., Xu, J., Hu, Q., Evaluation of antioxidant potential of Aloe vera (Aloe barbadensis Miller) extracts, J. Agric. Food Chem., 51, 7788- 7791 (2003).
• 27. Akev, N., Turkay, G., Can, A., Gurel, A., Yildiz, F., Yardibi, H., Ekiz, E.E., Uzun, H., Tumour preventive effect of Aloe vera leaf pulp lectin (Aloctin I) on Ehrlich ascites tumours in mice, Phytother. Res., 21, 1070-1075 (2007).