Alıç (crataegus monogyna) meyvesinden polifenol oksidaz enziminin kısmi saflaştırılması ve karakterizasyonu

Yıl 2018, Cilt: 22 Sayı: 3, 1048 - 1054, 01.06.2018

Ayşe Türkhan , Elif Duygu Kaya İbrahim Yılmaz

https://doi.org/10.16984/saufenbilder.334811

Cited By: 1

Öz

Bu
çalışmada, Alıç (crataegus monogyna) meyvesinden polifenol oksidaz (PFO) enzimi,
soğuk aseton çöktürmesi metodu kullanılarak kısmi olarak saflaştırılmış ve
karakterize edilmiştir. Enzimin optimum pH ve optimum sıcaklık değerleri
sırasıyla 5.0 ve 30°C olarak belirlenmiştir. Katekol substratı için Lineweaver-Burk
eğrisi ile maksimum reaksiyon hızı (V_maks) ve Michaelis–Menten
sabiti (K_m) değerleri sırası ile 5507 U/mL.dak ve 12.72 mM olarak
hesaplanmıştır. İnhibisyon çalışmaları polifenol oksidazın yaygın inhibitörleri
olan askorbik asit, sodyum metabisülfit ve benzoik asit ile yapılmış ve IC₅₀ değerleri sırasıyla 0.012
mM, 0.099 mM, 2.21 mM olarak tespit edilmiştir. Farklı fenolik substratlarla
yapılan substrat spesifikliği çalışmasında enzimin katekol substratına duyarlılığının
4-metil katekol ve L-tirosin substratlarına göre daha fazla olduğu tespit
edilmiştir. 

Anahtar Kelimeler

crataegus monogyna, polifenol oksidaz, soğuk aseton çöktürmesi, karakterizasyon, inhibisyon

The partial purification and characterization of polyphenol oxidase from hawthorn (crataegus monogyna)

Öz

In
this study, polyphenol oxidase from crataegus
monogyna fruit was partially
purified using cold aceton precipitation and characterized. The optimum pH and
optimum 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 (V_max) and the
Michaelis-Menten constant (K_m)
values for the catechol substrate were calculated to be 12.72 mM and 5507
U/mL.dak, respectively, using the Lineweaver-Burk plot. Inhibition studies were
performed for ascorbic acid, sodium metabisulphite and benzoic acid, which are
common inhibitors of the enzyme, and the IC₅₀
values for each inhibitor were determined to be 0.012 mM, 0.099 mM, 2.21
mM, respectively. The PPO were tested on different phenolic substrates,
including 4-methylcatechol, catechol, L-tyrosine. The highest activity was
observed in the presence of catechol for enzyme. 

Anahtar Kelimeler

crataegus monogyna, polyphenol oxidase, cold acetone precipitation, characterization, inhibition

Kaynakça

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