Purification and Characterization of Protease Enzyme from Oleander (Nerium oleander) Flowers of Different Colors

Year 2017, Volume: 1 Issue: 1, 21 - 26, 15.12.2017

Nazan Demir , Sila Nezahat Daşdemir Zehra Can

Abstract

ollected from Mugla countryside in June-September period. Protease catalyzes the hydrolysis of proteins to peptides and amino acids. It is one of the most important enzyme groups in both industrial and biochemical applications. Using the TPP method, the protease enzyme from oleander flowers (white, pink and red) was obtained in order of high efficiency. Optimum pH and optimum temperature for enzyme, KM and Vmax values for casein substrates were determined. SDS-PAGE was used to control the purity of the protease enzyme purified from oleander flowers. The molecular weight of protease enzyme purified from oleander flowers was determined by gel filtration chromatography to be white oleander 40,537 kea pink oleander 21,386 kDa and 22,516 kDa.

Keywords

Oleander (Nerium oleander), Purification, Protease, Characterization

References

• [1] Onat T, Emerk K. Temel Biyokimya [Basic Biochemistry]. İzmir: Saray Medikal Yayıncılık (1997).
• [2] Topal Ş, Pembeci C, Borcaklı M, Batum M, Çeltik Ö. Türki-ye’nin Tarımsal Mikoflorasının Endüstriyel Öneme Sahip Bazı Enzimatik Aktivitelerinin İncelenmesi-I: Amilaz, Proteaz, Lipaz. Turkish Journal of Biology (2000) 24):79–93.
• [3] Çalkı Ş. Bazı Su Ürünlerinde Proteolitik Enzim Aktiviteleri [Proteolytic Enzyme Activities in Some Aquatic Products]. Turkish Journal of Veterinary and Animal Sciences (1999) 23):385–390.
• [4] Demir Y, Alayli A., Yıldırım S, Demir N. Identification of Protease from Euphorbia Amygdaloides Latex and It’s Using in Cheese Producing. Biochemistry and Biotechnology (2005) 35):291–299.
• [5] Barrett AJ, Rawlings ND, Woessner JF. Hand-book of Proteo-lytic Enzymes. San Diego: Academic Press (1998).
• [6] Lecaille F, Kaleta. J., Brömme D. Human and parasitic papain-like cysteine proteases: their role in physiology and pathology and recent developments in inhibitor design. Chemical Reviews (2002) 102):4459.
• [7] Rawdkuen S, Chaiwut P, Pintathong P, Benjakul S. Three-phase partitioning of protease from Calotropis procera latex. Bio-chemical Engineering Journal (2010) 50(3):145–149. doi:10.1016/j.bej.2010.04.007.
• [8] Fadıloğlu S. Immobilization and characterization of ficin. Die Nahrung [Food] (2001) 45(2):143–146. doi:10.1002/1521-3803(20010401)45:2<143:AID-FOOD143>3.0.CO;2-8.
• [9] Laemmli UK. Cleavage of structural proteins during the as-sembly of the head of bacteriophage T4. Nature (1970) 227(5259):680–685.
• [10] Bradford MM. A rapid and sensitive method for the quantita-tion of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry (1976) 72):248–254.
• [11] Lehninger AL. Principles of biochemistry. New York: Worth Publishers Inc. (2013).
• [12] Hashim MM, Mingsheng D, Iqbal MF, Xiaohong C. Ginger rhizome as a potential source of milk coagulating cysteine pro-tease. Phytochemistry (2011) 72):458–464.
• [13] Nafi A, Foo HL, Jamilah B, Ghazali HM. Properties of proteo-lytic enzyme from ginger (Zingiber officinale Roscoe). Interna-tional Food Research Journal (2013) 20(1):363–368.
• [14] Demir Y, Güngör AA, Duran ED, Demir N. Cysteine protease (capparin) from capsules of caper (Capparis spinosa). Food Technology Biotechnology (2008) 46(3):286–291.