Bazı makrohalkalı tiyocrown eterlerin polifenol oksidaz enzimi üzerindeki inhibisyon etkilerinin araştırılması

Yıl 2018, , 539 - 546, 28.05.2018

Adem Ergün , Baki Çiçek

https://doi.org/10.25092/baunfbed.427644

Cited By: 1

Öz

Polifenol oksidaz (PPO) enzimi, meyvelerde, sebzelerde ve bazı hayvansal dokularda bolca bulunabilen, aktif bölgesinde bakır olan bir metalo enzimdir.  Enzimatik esmerleşme reaksiyonlarını katalizleyen bu enzimin, ticari değere sahip ürünlerde bolca bulunmasından dolayı inhibisyonu oldukça önem kazanmıştır.  Bunun için literatürde birçok çalışma mevcuttur.  Bu çalışmada, PPO enzimi, amonyum sülfat çöktürmesi ve diyaliz gibi ön saflaştırma işlemlerinden sonra, Sepharose 4B-l-tirozin -p-aminobenzoik asit afinite kromatografi jeli kullanılarak, Musa sapientum var. Cavendishii (Muz) meyvesinden saflaştırılmıştır.  Enzimin saflık kontrolü de SDS – PAGE ile yapılmıştır.  Daha sonra, bizim tarafımızdan esterleşme – halka kapama reaksiyonu sonucunda sentezlenen AE1, AE2 ve AE3 kodlu makrohalkalı tiyocrown eterlerin, saf olarak elde ettiğimiz polifenoloksidaz enzimi üzerindeki inhibisyon etkileri araştırılmıştır.  Tüm maddelerin PPO enzimini inhibe ettiği saptanmıştır.  Sonuçlara göre en etkili inhibitör AE3 olarak tespit edilmiştir. 

Anahtar Kelimeler

Tiyocrown eter, polifenol oksidaz (PPO), inhibisyon, Afinite kromatografisi

Investigation of the ınhibitory effects of some macrogenetic thiocrown ethers on polyphenol oxidase enzyme

Öz

The polyphenol oxidase (PPO) enzyme is a metallo enzyme that is copper in its active site, which can be found abundantly in fruits, vegetables and some animal tissues. Inhibition of this enzyme, which catalyzes the enzymatic browning reactions, is gaining in importance due to its abundance in commercial value products.  There are many studies in the literature for this.  In this study, PPO enzyme was purified using a Sepharose 4B-1-tyrosine-p-aminobenzoic acid affinity chromatography gel, after preliminary purification, such as ammonium sulfate precipitation and dialysis, from Musa sapientum var. Cavendishii (Banana).  Purity control of the enzyme was also performed by SDS - PAGE. We then investigated the inhibitory effects of the macrophilic thiocrown ethers AE1, AE2 and AE3 synthesized as a result of the esterification-ring closure reaction on the pure polyphenoloxidase enzyme.  All substances were found to inhibit PPO enzyme.  According to the results, the most effective inhibitor was identified as AE3.

Anahtar Kelimeler

Thiocrown ether, polyphenol oxidase (PPO), inhibition, affinity chromatography

Kaynakça

• Mayer, A.M., and Harel, E., Polyphenol Oxidases in Plants, Phytochemistry, 18, 2, 193-215, (1979).
• Friedman, M., Chemistry, Biochemistry, and dietary role of potato polyphenols, Journal of Agricultural and Food Chemistry, 45, 1 523-1540, (1997).
• Sekme, S., Çeşitli Mantarlarda Polifenol Oksidaz İndüksiyonunun İncelenmesi, Yüksek Lisans Tezi, Yıldız Teknik Universitesi Fen Bilimleri Enstitüsü, İstanbul, (2011).
• Gedikli, S., Çeşitli Makrofungus İzolatlarının Lakkaz Uretim Yetenekleri Açısından Değerlendirilmesi ve Dekolorizasyon Uygulamalarında Kullanılabilirliği, Yüksek Lisans Tezi, Eskisehir Osmangazi Universitesi Fen Bilimleri Enstitüsü, Eskişehir, (2008).
• Whitaker. J.R., Principles of Enzymology for the Food Sciences, Marcel 201, New York, Dekker, 22-24, (1972).
• Sarkar, J.M., Leonowicz, A. and Bollog. J.M., Immobilization of enzymes on clays and soils, Soil Biology and Biochemistry, 21 (2), 223-230, (1989).
• Harel, E., Mayer, A.M. and Shain, Y., Catechol oxidases from apples, their properties, subcellular location and inhibition, Plant Physiologoy, 17, 921, (1964).
• Tolbert, N.E., Activation of polyphenol oxidase of kloroplasts, Plant Pathology, 51, 234, (1973).
• Stephens, G.J. and Wood, R.K.S., Release of enzymes from cell walls by an endopectate-trans-eliminase, Nature, 251, 358, (1974).
• Padron, M.P., Lorano, J.A. and Gonsales, A.G., Properties of o- diphenol oxidoreductase from Musa cavendsha, Phytochemistry, 14, 1959, (1975).
• Arslan, O., Erzengin, M., Sinan, S. and Ozensoy, O., Purification of mulberry (Morus alba L.) polyphenol oxidase by affinity chromatography and investigation of its kinetic and electrophoretic properties, Food Chemistry, 88, 479-484, (2004).
• Spille, G.A., The Plant and Its Manufacture; Chemistry and Consumption of the Beverage, 1-38, (1997).
• Yue-Ming, J., Zauberman, G. and Fuchs, Y., Partial purification and some properties of polyphenol oxidase extracted from litchi fruit pericarp, Postharvest Biology and Technology, 10, 221-228, (1997).
• Mazzafera, P. and Robinson, S.P., Characterization of polyphenol oxidase in coffee, Phytochemistry, 55, 285-296, (2000).
• Shi, C., Dai, Y., Xu, X., Xie, Y. and Liu, Q., The purification of polyphenol oxidase from tobacco, Protein Expression and Purification, 24, 51-55, (2002).
• Aydemir, T., Partial purification and characterization of polyphenol oxidase from artichoke (Cynarascolymus L.) heads, Food Chemistry, 87, 59–67, (2004).
• Vaidya, B.K., Suthar, H.K., Kasture, S. and Nene, S., Purification of potatopolyphenol oxidase (PPO) by partitioning in aqueoustwo-phasesystem, Biochemical Engineering Journal, 28, 161–166, (2006).
• Waliszewski, K.N., Márquez, O. and Pardio, V.T., Quantification and characterisation of polyphenol oxidase from vanilla bean, Food Chemistry,117, 196–203, (2009).
• Guo, L., Ma, Y., Shi, J. and Xue, S., The purification and characterisation of polyphenoloxidase from gren bean (Phaseolusvulgaris L.), Food Chemistry, 117, 143–151, (2009).
• Franck C., Lammertyn, J., Ho, Q.T., Verbohen, P., Verlinden, B. and Nicolai, B. M., Browning disorders in pear fruit, Postharvest Biology and Technology, 43, 1–13, (2007).
• Alvarez-Parrilla, E., Rosa, L.A., Rodrigo-Garcia, J., Escobedo-Gonzalez, R., Mercado-Mercado, G., Moyers-Montoya, E., Vazquez-Flores, A. and Gonzalez-Aguilar, G.A., Dual effect of β-cyclodextrin (β-CD) on theinhibition of apple polyphenoloxidase by 4-hexylresorcinol (HR) and methyl jasmonate (MJ), Food Chemistry, 101, 1346–1356, (2007).
• Ünal, M.Ü. and Şener A., Two-year comparison of the biochemical properties of polyphenol oxidase from Turkish Alyanak apricot (Prunus armenica L.), Food Chemistry, 190, 741-747, (2016).
• Arslan, O., Temur, A. and Tozlu, I., Polyphenol oxidase from Malatya apricot (Prunus armeniaca L.), Journal of Agricultural and Food Chemistry, 46, 1239–1241, (1998).
• Doğan, S., Ayyıldız, Y., Doğan, M., Alan, Ü. and Diken, M.E., Characterisation of polyphenol oxidase from Melissa officinalis L. subsp. officinalis (lemon balm), Czech Journal of Food Science, 31, 2, 156-165, (2013).
• Doğan, S., Diken, M..E., Turhan, Y., Alan, Ü., Doğan, M. and Alkan, M., Characterisation and inhibition of Rosmarinus officinalis L. polyphenoloxidase., Eur Food Res Technol, 233, 293-301, (2011).
• Doğan, S., Diken, M.E., Alan, Ü., Yılmaz, B., Alkan, M. and Doğan, M., Some kinetic and inhibition properties of deepwater pink shrimp from aegean sea: ph, temperature, kinetic., inhibition, Advences in Food Sciences, 38, 155, (2016).
• Vamos-Vigyazo, L., Polyphenoloxidase and peroxidase in fruits and vegeatables, CRC Critcal Rewievs in Food Science and Nutrition, 15, 49-127, (1981).
• Frensch, V.K and Vogtle, F., Neuartige kronenether-lactone und -thiolactone. und ihre alkali- und erdalkaliion-komplexe, Tetrahedron Letters, 30, 2573, (1977).
• Alberts, A.H. and Cram, D.F., Syntheses and binding characteristics of macrocyclic systems containing one to three β-diketone units, Journal of Chemical Society, Chemical Communications, 28, 958, (1976).
• Pelissard, D. and Louis, R., Ligands macrocycliques pentadendates, Tetrahedron Letters., 45, 4589, (1972).
• Chiu, J.J., Grewal, R.S., Hart, H. and Ward, D.L., Cyclic-ketones via the reaction of dithiols with 1,3-dichloroacetone - an unexpected base-catalyzed rearrangement of alpha,alpha'-dithia ketones, Journal of Organic Chemistry, 58, 1553, (1993).
• Bradshow, J.S., Maas, G.E., Izatt, R.M. and Christensen, J.J., Synthetic macrocylic di- and tetraester compounds, Chemical Reviews, 79, 37, (1979).
• Izatt, R.M., Lab, J.D., Maas, G.E., Asay, R.E., Bradshaw, J.S. and Christensen, J.J., Asymmetric hydrogenation of α,β-dehydroamino acid residue in cyclic dipeptides, Journal of American Chemical Society, 99, 2365, (1977).
• Bradshaw, J.S., Hansen, L.D., Nielsen, S.F., Tompson, M.D., Reeder, R.N., Izatt, R.M. and Christensen, J.J., A new class of macrocyclic ether-ester ligands, Journal of Chemical Society, Chemical Communications, 21, 874-875, (1975).
• Bradshaw, J.S., Bishop, C.T., Nielsen, S.F., Asay, R.E., Mashidas, D.R., Flanders, E.D., Hansen, L.D., Izatt, R.M. and Christensen, J.J., Preparation of macrocyclic ether-esters, thioether-esters, and ether-thiolesters, Journal of Chemical Society Perkin Transactions 1, 23, 2505-2508, (1976).
• Asay, R.E., Bradshaw, J.S., Nielsen, S.F., Tompson, M.D., Snow, J.W., Mashidas, D.R.K., Izatt, R.M. and Christensen, J.J., The synthesis of novel macrocyclic multidentate compounds from dioxodioic acids, Journal of Heterocyclic Chemistry, 14, 85, (1977).
• Maas, G.M., Bradshaw, J.S., Izatt, R.M. and Christensen, J.J., Synthesis of a new series of macrocyclic polyether-diester ligands, Journal of Organic Chemistry, 42, 3937, (1977).
• Tompson, M.D., Bradshaw, J.S., Nielsen, S.F., Bishop, C.T., Cox, F.T., Fore, P.E., Maas, G.E., Izatt, R.M. and Christensen, J.J., The synthesis of some substituted macrocyclic ether-ester compounds, Tetrahedron, 33, 3317, (1977).
• Fore, P.E., Bradshaw, J.S. and Nielsen, S.F., The synthesis of macrocyclic ether esters, thioetherbesters, and ether thiolesters with the oxalyl moiety, Journal of Heterocyclic Chemistry, 15, 269, (1978).
• Bradshaw J.S. and Tompson, M.D., Synthesis of macrocyclic polyether-diester compounds with an aromatic subcyclic unit, Journal of Organic Chemistry, 43, 2456, (1978).
• Izatt, R.M., Lamb, J.D., Asay, R.E., Maas, G.E., Bradshaw, J.S., Christensen, J.J. and Moore, S.S., Unusual stability characteristics in methanol of the complexes of a new pyridibe-substituted cyclic polyether-ester compound with Na+, K+, Ag+ and Ba2+ comparison with oxygen, sulfur, nitrogen analogues, Journal of American Chemical Society, 99, 6134, (1977).
• Matsushima, K., Synthesis of novel macrocyclic ether-ester compounds via the intramolecular cyclization of oligoethylene glycol monocarboxymethyl ethers, Tetrahedron Lettets, 20, 3445, (1979).
• Edema, J.J., Buter, J., Kellogg, R.M., Spek, A.L. and Bolhuis, F.V., Intra- versus inter-molecular azine formation in thiocrown ether chemistry, Journal of the Chemical Society, Chemical Communications, 21, 1558, (1992).
• Çiçek, B., Ergün, A. and Gençer, N., Synthesis and evaluation in vitro effects of some macrocyclic thiacrown ethers on erythrocyte carbonic anhydrase I and II, Asian Journal of Chemistry, 24, 7, 3729-3731, (2012).
• Laemelli, D.K., Cleavege of structural ptoteins during in assembly of thehead of bacteriophage, Nature, 227-680, (1970).
• Bradford, M., A rapid and sensitive method for the quantitaion of microgram quantites of protein utilizing the principle of protein-dye binding, Analytical Biochemistry, 72, 248, (1976).
• Espin J.C., Morales M., Varon R., Tudela J. and Garcia-Canovas F., Acontinuous spectrophotometric method for determining the monophenolase and diphenolase activities of apple polyphenol oxidase, Analytical Biochemistry, 43, 2807–2812, (1995).
• Sayaverde-Soto L.A. and Montgomery M.W., Inhibition of polyphenol oxidase by sülfte, Journal of Food Science, 51, 1531–1535, (1986).
• Embs R.J. and Markakis P.T., Mechanism of sulphite inhibition of browning caused by polyphenol oxidase. Journal of Food Science., 30, 753–758, (1965).
• Schwimmer S. and Schwimmer S., Source Book of Food Enzymology. AVI Publishing, Westport, 274, 267, (1981).
• Chilaka F.C., Eze S., Anyadiegwu C. and Uvere P.O., Browning in processed yams: peroxidase or polyphenol oxidase, Journal of the Science of Food and Agriculture, 82, 899–903, (2002).