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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

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 (Vmaks) ve Michaelis–Menten
sabiti (Km) 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

Kaynakça

  • [1] R. Yoruk and M. R. Marshall, “Physicochemical properties and function of plant polyphenoloxidase:a review,” Journal of Food Biochemistry, 27, 361–422, 2003.
  • [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.
  • [3] H. S. Mason, “Chemistry of melanin Mechanism of oxidation of 3,4-dihydroxyphenylalanine by tyrosinase,” Journal of Biological Chemistry, 72, 83–99, 1948.
  • [4] G. Prota, “Progress in the chemistry of melanins and related metabolites,” Medicinal Research Review, 8, 525–556, 1988.
  • [5] P. R. Mathewson, “Enzymes,” Eagen Press Handbook Series, 37-38, 2000.
  • [6] D. J. Mcweeny, “The chemistry of non-enzymatic browning in foods and its control by sulphites,” Journal of the Science of Food and Agriculture, 25, 735, 1974.
  • [7] T. P. Labuza, J. H. Lillemo, and P. S. Taoukis, “Inhibition of polyphenol oxidase by proteolytic enzymes,” Fruit Processing, 2, 9-13, 1992.
  • [8] A. Batu, A. Çağlar, Ö. Emrem, and B. Çeliker, “Alıç Pekmezi Üretimi,” Gıda Teknolojileri Dergisi, 2, 45-51, 2007.
  • [9] E. Middleton, “ The flavonoids,” Trends in Pharmaceutical Sciences, 5, 335-338, 1984.
  • [10] N. C. Cook and S. Samman, “Flavonoids-chemistry, metabolism, cardio protective effects, and dietary sources,” The Journal of Nutritional Biochemistry, 7, 66–761, 996.
  • [11] T. Aydemir, “Partial purification and characterization of polyphenol oxidase from artichoke (cynara scolymus L.) heads,” Food Chemistry, 87, 59–67, 2004.
  • [12] O. H. Lowry, N. J. Rosebrough, A. L. Farr, and R. J. Randall, “Protein Measurement With The Folin Phenol Reagent,” Journal of Biological Chemistry, 193, 265-275, 1951.
  • [13] J. C. Espin, M. Morales, R. Varon, J. Tudela, and F. Garcia-Canovas, “A continuous spectrophotometric method for determining the monophenolase and diphenolase activities of apple polyphenol oxidase,” Analytical Biochemistry, 43, 2807-2812, 1995.
  • [14] M. A. M. Galeazzi and V. C. J. Sgarbieri, “Substrate Specificity and İnhibition of Polyphenoloxidase from a Dwarf Variety of Banana (musa cavendishii, L.),” Journal Of Food Science, 46, 1404-1406, 1981.
  • [15] A. Colak, A. Özen, B. Dincer, S. Güner, and A. F. Ayaz, “Diphenolases from Two Cultivars of Cherry Laurel (laurocerasus officinalis roem.) Fruits at Early Stage of Maturation,” Food Chemistry, 90, 801-807, 2005.
  • [16] B. Dincer, A. Colak, N. Aydin, A. Kadioglu, and S. Güner, “Characterization of Polyphenoloxidase from Medlar Fruits (mespilus germanica L. rosaceae),” Food Chemistry, 77, 1-7, 2002.
  • [17] H. Lineweaver and D. Burk, “The Determination of Enzyme Dissociation Constant,” Journal of American Chemical Society, 56, 658-661, 1934.
  • [18] İ. Kuyumcu, “Yabani Ve Yenilebilir Bir Mantar Olan Lactarıus Eucalyptı O. K. Mıll & R. N. Hılton’ dan Polifenol Oksidazın Saflaştırılması Ve Karakterizasyonu,” Karadeniz Teknik Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, 2014.
  • [19] F. Öz, A. Colak, A. Özel, N. Sağlam-Ertunga, and E. Sesli, “Purification and Characterization of A Mushroom Polyphenol Oxidase and Its Activity in Organic Solvents,” Journal of Food Biochemistry, 37, 36–44, 2013.
  • [20] A. A. Khan, S. Akhtar, and Q. Husain, “Direct immobilization of polyphenol oxidases on celite 545 from ammonium sulphate fractionated proteins of potato (solanum tuberosum),” Journal of Molecular Catalysis B: Enzymatic, 40, 58-63, 2006.
  • [21] K. Bravo and E. Osorio, “Characterization of polyphenol oxidase from Cape gooseberry (physalis peruviana L.) fruit,” Food Chemistry, 197, 185–190, 2016.
  • [22] M. Erat, Y. N. Şahin, G. Aksoy, and A. Demirkol, “Partial characterization of polyphenoloxidase from a hybridized wheat (triticum aestivum L.),” European Food Research and Technology, 231, 899–905, 2010.
  • [23] M. Oktay, I. Küfrevioğlu, I. Kocaçalışkan, and H. Şakırolu, “Polyphenol oxidase from Amasya apple,” Journal of Food Science, 60, 494–496, 1995.
  • [24] S. Doğan and Ü. Salman, “Partial characterization of lettuce (lactuca sativa L.) polyphenol oxidase,” European Food Research and Technology, 226, 93–103, 2007.
  • [25] H. Şakiroğlu, A. E. Öztürk, A. E. Pepe, and M. Erat, “Some kinetic properties of polyphenol oxidase obtained from dill (anethum graveolens),” Journal of Enzyme Inhibition and Medicinal Chemistry, 23, 380–385, 2008.
  • [26] F. Ülker-Yerlitürk, O. Arslan, S. Sınan, N. Gencer and Ö. Özensoy, “Characterızatıon Of Polyphenoloxıdase From Wıld Pear (pyrus elaegrıfolıa),” Journal of Food Biochemistry, 32, 368–383, 2008.
  • [27] S. Doğan, Y. Turan, H. Ertürk, and O. Arslan, “Characterization and Purification of Polyphenol Oxidase from Artichoke (cynara scolymus L.),” Journal of Agricultural and Food Chemistry, 53, 776-785, 2005.
  • [28] A. W. R. Ng and C. W. Wong, “Partial purification and characterization of polyphenol oxidase from round brinjal (S. melongena var. depressum),” International Food Research Journal, 22, 826-831, 2015.

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

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

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 (Vmax) and the
Michaelis-Menten constant (Km)
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

  • [1] R. Yoruk and M. R. Marshall, “Physicochemical properties and function of plant polyphenoloxidase:a review,” Journal of Food Biochemistry, 27, 361–422, 2003.
  • [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.
  • [3] H. S. Mason, “Chemistry of melanin Mechanism of oxidation of 3,4-dihydroxyphenylalanine by tyrosinase,” Journal of Biological Chemistry, 72, 83–99, 1948.
  • [4] G. Prota, “Progress in the chemistry of melanins and related metabolites,” Medicinal Research Review, 8, 525–556, 1988.
  • [5] P. R. Mathewson, “Enzymes,” Eagen Press Handbook Series, 37-38, 2000.
  • [6] D. J. Mcweeny, “The chemistry of non-enzymatic browning in foods and its control by sulphites,” Journal of the Science of Food and Agriculture, 25, 735, 1974.
  • [7] T. P. Labuza, J. H. Lillemo, and P. S. Taoukis, “Inhibition of polyphenol oxidase by proteolytic enzymes,” Fruit Processing, 2, 9-13, 1992.
  • [8] A. Batu, A. Çağlar, Ö. Emrem, and B. Çeliker, “Alıç Pekmezi Üretimi,” Gıda Teknolojileri Dergisi, 2, 45-51, 2007.
  • [9] E. Middleton, “ The flavonoids,” Trends in Pharmaceutical Sciences, 5, 335-338, 1984.
  • [10] N. C. Cook and S. Samman, “Flavonoids-chemistry, metabolism, cardio protective effects, and dietary sources,” The Journal of Nutritional Biochemistry, 7, 66–761, 996.
  • [11] T. Aydemir, “Partial purification and characterization of polyphenol oxidase from artichoke (cynara scolymus L.) heads,” Food Chemistry, 87, 59–67, 2004.
  • [12] O. H. Lowry, N. J. Rosebrough, A. L. Farr, and R. J. Randall, “Protein Measurement With The Folin Phenol Reagent,” Journal of Biological Chemistry, 193, 265-275, 1951.
  • [13] J. C. Espin, M. Morales, R. Varon, J. Tudela, and F. Garcia-Canovas, “A continuous spectrophotometric method for determining the monophenolase and diphenolase activities of apple polyphenol oxidase,” Analytical Biochemistry, 43, 2807-2812, 1995.
  • [14] M. A. M. Galeazzi and V. C. J. Sgarbieri, “Substrate Specificity and İnhibition of Polyphenoloxidase from a Dwarf Variety of Banana (musa cavendishii, L.),” Journal Of Food Science, 46, 1404-1406, 1981.
  • [15] A. Colak, A. Özen, B. Dincer, S. Güner, and A. F. Ayaz, “Diphenolases from Two Cultivars of Cherry Laurel (laurocerasus officinalis roem.) Fruits at Early Stage of Maturation,” Food Chemistry, 90, 801-807, 2005.
  • [16] B. Dincer, A. Colak, N. Aydin, A. Kadioglu, and S. Güner, “Characterization of Polyphenoloxidase from Medlar Fruits (mespilus germanica L. rosaceae),” Food Chemistry, 77, 1-7, 2002.
  • [17] H. Lineweaver and D. Burk, “The Determination of Enzyme Dissociation Constant,” Journal of American Chemical Society, 56, 658-661, 1934.
  • [18] İ. Kuyumcu, “Yabani Ve Yenilebilir Bir Mantar Olan Lactarıus Eucalyptı O. K. Mıll & R. N. Hılton’ dan Polifenol Oksidazın Saflaştırılması Ve Karakterizasyonu,” Karadeniz Teknik Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, 2014.
  • [19] F. Öz, A. Colak, A. Özel, N. Sağlam-Ertunga, and E. Sesli, “Purification and Characterization of A Mushroom Polyphenol Oxidase and Its Activity in Organic Solvents,” Journal of Food Biochemistry, 37, 36–44, 2013.
  • [20] A. A. Khan, S. Akhtar, and Q. Husain, “Direct immobilization of polyphenol oxidases on celite 545 from ammonium sulphate fractionated proteins of potato (solanum tuberosum),” Journal of Molecular Catalysis B: Enzymatic, 40, 58-63, 2006.
  • [21] K. Bravo and E. Osorio, “Characterization of polyphenol oxidase from Cape gooseberry (physalis peruviana L.) fruit,” Food Chemistry, 197, 185–190, 2016.
  • [22] M. Erat, Y. N. Şahin, G. Aksoy, and A. Demirkol, “Partial characterization of polyphenoloxidase from a hybridized wheat (triticum aestivum L.),” European Food Research and Technology, 231, 899–905, 2010.
  • [23] M. Oktay, I. Küfrevioğlu, I. Kocaçalışkan, and H. Şakırolu, “Polyphenol oxidase from Amasya apple,” Journal of Food Science, 60, 494–496, 1995.
  • [24] S. Doğan and Ü. Salman, “Partial characterization of lettuce (lactuca sativa L.) polyphenol oxidase,” European Food Research and Technology, 226, 93–103, 2007.
  • [25] H. Şakiroğlu, A. E. Öztürk, A. E. Pepe, and M. Erat, “Some kinetic properties of polyphenol oxidase obtained from dill (anethum graveolens),” Journal of Enzyme Inhibition and Medicinal Chemistry, 23, 380–385, 2008.
  • [26] F. Ülker-Yerlitürk, O. Arslan, S. Sınan, N. Gencer and Ö. Özensoy, “Characterızatıon Of Polyphenoloxıdase From Wıld Pear (pyrus elaegrıfolıa),” Journal of Food Biochemistry, 32, 368–383, 2008.
  • [27] S. Doğan, Y. Turan, H. Ertürk, and O. Arslan, “Characterization and Purification of Polyphenol Oxidase from Artichoke (cynara scolymus L.),” Journal of Agricultural and Food Chemistry, 53, 776-785, 2005.
  • [28] A. W. R. Ng and C. W. Wong, “Partial purification and characterization of polyphenol oxidase from round brinjal (S. melongena var. depressum),” International Food Research Journal, 22, 826-831, 2015.
Toplam 28 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Kimya Mühendisliği
Bölüm Araştırma Makalesi
Yazarlar

Ayşe Türkhan

Elif Duygu Kaya Bu kişi benim

İbrahim Yılmaz Bu kişi benim

Yayımlanma Tarihi 1 Haziran 2018
Gönderilme Tarihi 21 Ağustos 2017
Kabul Tarihi 28 Mayıs 2018
Yayımlandığı Sayı Yıl 2018 Cilt: 22 Sayı: 3

Kaynak Göster

APA Türkhan, A., Kaya, E. D., & Yılmaz, İ. (2018). Alıç (crataegus monogyna) meyvesinden polifenol oksidaz enziminin kısmi saflaştırılması ve karakterizasyonu. Sakarya University Journal of Science, 22(3), 1048-1054. https://doi.org/10.16984/saufenbilder.334811
AMA Türkhan A, Kaya ED, Yılmaz İ. Alıç (crataegus monogyna) meyvesinden polifenol oksidaz enziminin kısmi saflaştırılması ve karakterizasyonu. SAUJS. Haziran 2018;22(3):1048-1054. doi:10.16984/saufenbilder.334811
Chicago Türkhan, Ayşe, Elif Duygu Kaya, ve İbrahim Yılmaz. “Alıç (crataegus Monogyna) Meyvesinden Polifenol Oksidaz Enziminin kısmi saflaştırılması Ve Karakterizasyonu”. Sakarya University Journal of Science 22, sy. 3 (Haziran 2018): 1048-54. https://doi.org/10.16984/saufenbilder.334811.
EndNote Türkhan A, Kaya ED, Yılmaz İ (01 Haziran 2018) Alıç (crataegus monogyna) meyvesinden polifenol oksidaz enziminin kısmi saflaştırılması ve karakterizasyonu. Sakarya University Journal of Science 22 3 1048–1054.
IEEE A. Türkhan, E. D. Kaya, ve İ. Yılmaz, “Alıç (crataegus monogyna) meyvesinden polifenol oksidaz enziminin kısmi saflaştırılması ve karakterizasyonu”, SAUJS, c. 22, sy. 3, ss. 1048–1054, 2018, doi: 10.16984/saufenbilder.334811.
ISNAD Türkhan, Ayşe vd. “Alıç (crataegus Monogyna) Meyvesinden Polifenol Oksidaz Enziminin kısmi saflaştırılması Ve Karakterizasyonu”. Sakarya University Journal of Science 22/3 (Haziran 2018), 1048-1054. https://doi.org/10.16984/saufenbilder.334811.
JAMA Türkhan A, Kaya ED, Yılmaz İ. Alıç (crataegus monogyna) meyvesinden polifenol oksidaz enziminin kısmi saflaştırılması ve karakterizasyonu. SAUJS. 2018;22:1048–1054.
MLA Türkhan, Ayşe vd. “Alıç (crataegus Monogyna) Meyvesinden Polifenol Oksidaz Enziminin kısmi saflaştırılması Ve Karakterizasyonu”. Sakarya University Journal of Science, c. 22, sy. 3, 2018, ss. 1048-54, doi:10.16984/saufenbilder.334811.
Vancouver Türkhan A, Kaya ED, Yılmaz İ. Alıç (crataegus monogyna) meyvesinden polifenol oksidaz enziminin kısmi saflaştırılması ve karakterizasyonu. SAUJS. 2018;22(3):1048-54.
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