TY - JOUR T1 - Alıç (crataegus monogyna) meyvesinden polifenol oksidaz enziminin kısmi saflaştırılması ve karakterizasyonu TT - The partial purification and characterization of polyphenol oxidase from hawthorn (crataegus monogyna) AU - Türkhan, Ayşe AU - Kaya, Elif Duygu AU - Yılmaz, İbrahim PY - 2018 DA - June Y2 - 2018 DO - 10.16984/saufenbilder.334811 JF - Sakarya University Journal of Science JO - SAUJS PB - Sakarya University WT - DergiPark SN - 2147-835X SP - 1048 EP - 1054 VL - 22 IS - 3 LA - tr AB - Buçalışmada, Alıç (crataegus monogyna) meyvesinden polifenol oksidaz (PFO) enzimi,soğuk aseton çöktürmesi metodu kullanılarak kısmi olarak saflaştırılmış vekarakterize edilmiştir. Enzimin optimum pH ve optimum sıcaklık değerlerisırasıyla 5.0 ve 30°C olarak belirlenmiştir. Katekol substratı için Lineweaver-Burkeğrisi ile maksimum reaksiyon hızı (Vmaks) ve Michaelis–Mentensabiti (Km) değerleri sırası ile 5507 U/mL.dak ve 12.72 mM olarakhesaplanmıştır. İnhibisyon çalışmaları polifenol oksidazın yaygın inhibitörleriolan askorbik asit, sodyum metabisülfit ve benzoik asit ile yapılmış ve IC50 değerleri sırasıyla 0.012mM, 0.099 mM, 2.21 mM olarak tespit edilmiştir. Farklı fenolik substratlarlayapılan substrat spesifikliği çalışmasında enzimin katekol substratına duyarlılığının4-metil katekol ve L-tirosin substratlarına göre daha fazla olduğu tespitedilmiştir. KW - crataegus monogyna KW - polifenol oksidaz KW - soğuk aseton çöktürmesi KW - karakterizasyon KW - inhibisyon N2 - Inthis study, polyphenol oxidase from crataegusmonogyna fruit was partiallypurified using cold aceton precipitation and characterized. The optimum pH andoptimum temperature of the enzyme were found to be 5.0 and 30°C, respectively,in the presence of catechol as a substrate. The maximum reaction rate (Vmax) and theMichaelis-Menten constant (Km)values for the catechol substrate were calculated to be 12.72 mM and 5507U/mL.dak, respectively, using the Lineweaver-Burk plot. Inhibition studies wereperformed for ascorbic acid, sodium metabisulphite and benzoic acid, which arecommon inhibitors of the enzyme, and the IC50values for each inhibitor were determined to be 0.012 mM, 0.099 mM, 2.21mM, respectively. The PPO were tested on different phenolic substrates,including 4-methylcatechol, catechol, L-tyrosine. The highest activity wasobserved in the presence of catechol for enzyme. CR - [1] R. Yoruk and M. R. Marshall, “Physicochemical properties and function of plant polyphenoloxidase:a review,” Journal of Food Biochemistry, 27, 361–422, 2003. CR - [2] Y. Kolcuoğlu, “Purification and comparative characterization of monophenolase and diphenolase activities from a wild edible mushroom (macrolepiota gracilenta),” Process Biochemistry, 47, 2449–2454, 2012. 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UR - https://doi.org/10.16984/saufenbilder.334811 L1 - https://dergipark.org.tr/en/download/article-file/484465 ER -