Purification and Characterization of Glutathione Reductase Enzyme from Arum Maculatum Leaf

Anahtar Kelimeler:

Glutathione reductase, Arum maculatum, Purification, Enzyme, Characterization

Purification and Characterization of Glutathione Reductase Enzyme from Arum Maculatum Leaf

Arum species grow in temperate and Mediterranean climates and have been used for hundreds of years for food and medicinal purposes, although they are highly toxic if not cooked using proper techniques. Glutathione reductase (GR) is a member of the pyridine nucleotide disulfide oxidoreductase family of flavoenzymes that catalyzes the reduction of glutathione disulfide (GSSG) to reduced GSH using NADPH or NADH. In this study, GR enzyme was characterized by partial purification processes including homogenate preparation, ammonium sulfate precipitation and dialysis from the leaf of Arum maculatum plant. The highest enzyme activity was found at 40-60% saturation range. Optimum ionic strength, pH and substrate concentration were investigated for GR enzyme from A. maculatum leaf. As a result of the study, these values were found to be 150 mM potassium phosphate buffer, pH 7.00, and 0.18 mM, respectively. The GR enzyme was partially purified from the leaf of the A. maculatum with a specific activity of 1.640 EU mg-1 in 34.9% yield, 1.108-fold. This study is the first study in terms of purification and characterization of GR enzyme from A. maculatum leaf.

Keywords:

Glutathione reductase, Arum maculatum, Purification, Enzyme, Characterization,

PDF

___

Acan NL, Tezcan EF. 1989. Sheep brain glutathione reductase: purification and general properties. FEBS Letters, 250(1): 72-74.
Ali A. 2008. Comparison of protein values from seven wild edible plants of Iran. African J Food Sci, 2(6): 073-076.
Alscher, R. G. 1989. Biosynthesis and antioxidant function of glutathione in plants. Physiol Plantarum, 77(3): 457-464.
Baytop T. 1999. Türkiye'de bitkiler ile tedavi: geçmişte ve bugün. Nobel Tıp Kitabevleri, İstanbul, Türkiye, pp: 480.
Bilir G. 2017. Soya bitkisinden (glycine max l.) glutatyon redüktaz enziminin saflaştırılması ve bazı ağır metallerin inhibisyon kinetiğinin incelenmesi. MSc thesis, Ondokuz Mayıs Üniversitesi, Fen Bilimleri Enstitüsü, Samsun, Türkiye, pp: 61.
Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochem, 72(1-2): 248-254.
Carlberg I, Mannervik B. 1975. Purification and characterization of the flavoenzyme glutathione reductase from rat liver. J Biol Chem, 250(14): 5475-5480.
Ceylan F, Sahingoz SA. 2022. Using ethnobotanical plants in food preparation: Cuckoo pint (Arum maculatum L.). Inter J Gastronomy Food Sci, 2022: 100529.
Couto N, Wood J, Barber J. 2016. The role of glutathione reductase and related enzymes on cellular redox homoeostasis network. Free Radical Biol Med, 95: 27-42.
Çakmak R, Durdagi S, Ekinci D, Şentürk M, Topal G. 2011. Design, synthesis and biological evaluation of novel nitroaromatic compounds as potent glutathione reductase inhibitors. Bioorganic Med Chem Letters, 21(18): 5398-5402.
Çolak F, Savaroğlu F, İlhan S. 2009. Antibacterial and antifungal activities of Arum maculatum L. leaves extracts. J Applied Biol Sci, 3(3): 13-16.
Dayisoylu KS. 2010. Changes of antioxidant activity in different forms and meal of Arum maculatum in Kahramanmaras province from Turkey. Asian J Chem, 22(8): 6595.
Erat M, Demir H, Sakiroglu H. 2005. Purification of glutathione reductase from chicken liver and investigation of kinetic properties. Applied Biochem Biotechnol, 125(2): 127-138.
Erat M, Sakiroglu H, Ciftci M. 2003. Purification and characterization of glutathione reductase from bovine erythrocytes. Preparat Biochem Biotechnol, 33(4): 283-300.
Farahmandfar R, Esmaeilzadeh Kenari R, Asnaashari M, Shahrampour D, Bakhshandeh T. 2019. Bioactive compounds, antioxidant and antimicrobial activities of Arum maculatum leaves extracts as affected by various solvents and extraction methods. Food Sci Nutri, 7(2): 465-475.
Foyer CH, Noctor G. 2005. Oxidant and antioxidant signalling in plants: a re‐evaluation of the concept of oxidative stress in a physiological context. Plant Cell Environ, 28(8): 1056-1071.
Gibernau M, Macquart D, Przetak G. 2004. Pollination in the genus Arum–a review. Aroideana, 27: 148-166.
Gill SS, Anjum NA, Hasanuzzaman M, Gill R, Trivedi DK, Ahmad I, Pereira E, Tuteja N. 2013. Glutathione and glutathione reductase: a boon in disguise for plant abiotic stress defense operations. Plant Physiol Biochem, 70: 204-212.
Kaur A, Tyagi S, Singh K, Upadhyay SK. 2022. Exploration of glutathione reductase for abiotic stress response in bread wheat (Triticum aestivum L.). Plant Cell Reports, 41(3): 639-654.
Kocaoğlu E, Talaz O, Çavdar H, Şentürk M, Supuran CT, Ekinci D. 2019. Determination of the inhibitory effects of N-methylpyrrole derivatives on glutathione reductase enzyme. J Enzyme Inhib Med Chem, 34(1): 51-54.
Lascano H, Casano L, Melchiorre M, Trippi V. 2001. Biochemical and molecular characterisation of wheat chloroplastic glutathione reductase. Biol Plantarum, 44(4): 509-516.
Liu HJ, Wang X, Yang ZL, Ren LL, Qian TT. 2020. Identification and biochemical characterization of the glutathione reductase family from Populus trichocarpa. Plant Sci, 294: 110459.
Madamanchi NR, Anderson JV, Alscher RG, Cramer CL, Hess JL. 1992. Purification of multiple forms of glutathione reductase from pea (pisum sativum l.) seedlings and enzyme levels in ozone-fumigated pea leaves. Plant Physiol, 100(1): 138-145.
Mahan JR, Burke JJ. 1987. Purification and characterization of glutathione-reductase from corn mesophyll chloroplasts. Physiol Plantarum, 71(3): 352-358.
Marc EB, Nelly A, Annick DD, Frederic D. 2008. Plants used as remedies antirheumatic and antineuralgic in the traditional medicine of Lebanon. J Ethnopharmacol, 120(3): 315-334.
Meister A, Anderson ME. 1983. Glutathione. Annual Rev Biochem, 52(1): 711-760.
Ogus IH, Ozer N. 1998. Purification of NADPH-free glutathione disulfide reductase from human erythrocytes. Protein Expr Purif, 13(1): 41-44.
Raju KP, Goel K, Anandhi D, Pandit VR, Surendar R, Sasikumar M. 2018. Wild tuber poisoning: Arum maculatum–A rare case report. Inter J Crit illness Injury Sci, 8(2): 111.
Romero‐Puertas MC, Corpas FJ, Sandalio LM, Leterrier M, Rodríguez‐Serrano M, Del Río LA, Palma JM. 2006. Glutathione reductase from pea leaves: response to abiotic stress and characterization of the peroxisomal isozyme. New Phytol, 170(1): 43-52.
Senturk M, Kufrevioglu OI, Ciftci M. 2008. Effects of some antibiotics on human erythrocyte glutathione reductase: an in vitro study. J Enzyme Inhibit Med Chem, 23(1): 144-148.
Senturk M, Kufrevioglu OI, Ciftci M. 2009. Effects of some analgesic anaesthetic drugs on human erythrocyte glutathione reductase: an in vitro study. J Enzyme Inhibit Med Chem, 24(2): 420-424.
Smith IK, Vierheller TL, Thorne CA. 1988. Assay of glutathione reductase in crude tissue homogenates using 5, 5′-dithiobis (2-nitrobenzoic acid). Analytical Biochem, 175(2): 408-413.
Şentürk E, Şentürk M. 2020. Investigation of Some Corticosteroids as Glutathione Reductase Inhibitor. Inter J Secondary Metabol, 7(2): 119-125.
Şentürk M, Talaz O, Ekinci D, Çavdar H, Küfrevioğlu Öİ. 2009. In vitro inhibition of human erythrocyte glutathione reductase by some new organic nitrates. Bioorganic Med Chem Letters, 19(13): 3661-3663.
Tekman B, Ozdemir H, Senturk M, Ciftci M. 2008. Purification and characterization of glutathione reductase from rainbow trout (Oncorhynchus mykiss) liver and inhibition effects of metal ions on enzyme activity. Comp Biochem Physiol C Toxicol Pharmacol, 148(2): 117-121.
Tetik F, Civelek S, Cakilcioglu U. 2013. Traditional uses of some medicinal plants in Malatya (Turkey). J Ethnopharmacol, 146(1): 331-346.
Townsend DM, Tew KD, Tapiero H. 2003. The importance of glutathione in human disease. Biomed Pharmacotherap, 57(3): 145-155.
Ulusu G, Erat M, Ciftci M, Sakiroglu H, Bakan E. 2005. Purification and characterization of glutathione reductase from sheep liver. Turkish J Vet Anim Sci, 29(5): 1109-1117.
Uzun E, Sariyar G, Adsersen A, Karakoc B, Ötük G, Oktayoglu E, Pirildar S. 2004. Traditional medicine in Sakarya province (Turkey) and antimicrobial activities of selected species. J Ethnopharmacol, 95(2-3): 287-296.
Wang X, Qian TT, Zeng QY. 2013. Biochemical characterization of larix kaempferi glutathione reductase. Forest Res, 26(S1): 25-32.
Willmore WG, Storey KB. 2007. Purification and properties of glutathione reductase from liver of the anoxia-tolerant turtle, Trachemys scripta elegans. Molec Cellular Biochem, 297(1-2): 139-149.
Wingsle G. 1989. Purification and characterization of glutathione reductase from Scots pine needles. Physiol Plantarum, 76(1): 24-30.
Wu TM, Lin WR, Kao YT, Hsu YT, Yeh CH, Hong CY, Kao CH. 2013. Identification and characterization of a novel chloroplast/mitochondria co-localized glutathione reductase 3 involved in salt stress response in rice. Plant Molec Biol, 83(4-5): 379-390.
Yurt NŞ, Eda T, Çubukçu M. 2019. Nivik otu zehirlenmesi: Arum maculatum-Bir olgu sunumu. Ankara Med J, 19(4): 796-799.