ISIL İŞLEMİN ELMA SUYUNDA ENZİM AKTİVİTESİ VE FENOLİK BİLEŞİKLERE ETKİSİ: FTIR VE HPLC ÇALIŞMASI

Öz

Bu çalışmada farklı sıcaklık (40, 50, 60, 70 ve 80 °C) ve sürelerde (5, 10, 15, 20, 25, 30 dakika) uygulanan ısıl işlemin elma suyunda Polifenol oksidaz (PPO) ve Peroksidaz (POD) enzim aktivitesi ile fenolik bileşikler üzerine etkisi incelenmiş ve fenolik bileşiklerdeki değişim FTIR spektroskopisi ve HPLC kullanılarak belirlenmiştir. Enzim inaktivasyonu genellikle artan sıcaklık ve zamanla artmıştır. PPO enziminde 80 °C’de 20 dakika sonunda % 99 inaktivasyon sağlanırken, POD enziminde 80 °C’de 20 dakika sonunda % 93,29 inaktivasyon sağlanmıştır. Genellikle 70 ve 80 °C sıcaklıkta toplam fenolik ve antioksidan aktivitede artış belirlenmiştir. Bu artış, enzim inaktivasyonu ile ilişkilendirilmiştir. 80 °C’ de 20 dakika ısıl işlemden sonra toplam fenolik madde miktarı 668,41 ± 12,70 mg GAE/kg yaş ağırlık, EC50 değeri 24,28 ± 0,69 mg/ml olarak belirlenmiştir. HPLC ile elma suyunda belirlenen fenolik bileşikler kateşin, klorojenik asit, epikateşin ve kamferol olmuştur. 80 °C’ de 20 dakikada gerçekleştirilen ısıl işlem sonunda fenolik bileşiklerin miktarında artış belirlenmiştir. FTIR spektrumları incelendiğinde spesifik bantların epikateşin ve klorojenik asitle uyumlu olduğu ve bulunan sonuçların HPLC ile paralellik gösterdiği belirlenmiştir. Ayrıca FTIR spektroskopisi ile fenolik bileşiklerde bir değişim gözlenmemiştir.

Anahtar Kelimeler

• Elma suyu
• Isıl işlem
• Fenolik Bileşik
• FTIR
• HPLC

EFFECT OF THERMAL TREATMENT ON ENZYME ACTIVITY AND PHENOLIC COMPOUNDS IN APPLE JUICE: FTIR AND HPLC STUDY

Öz

In this study, the effect of thermal treatment with different temperatures (40, 50, 60, 70, 80 ° C) and times (5, 10, 15, 20, 25, 30 minutes) on Polyphenol oxidase(PPO) and Peroxidase (POD) and phenolic compounds in apple juice was investigated and the change in phenolic compounds was determined by FTIR spectroscopy and HPLC. Enzyme inactivation generally increased with increasing temperature and time. In PPO 99 % and in POD 93.29 % inactivation was achieved after 20 minutes at 80 °C. Generally, an increase was determined at 70 and 80 °C in total phenolic content (TPC) and antioxidant activity. This increase has been associated with enzyme inactivation. After 20 minutes at 80 °C, TPC was determined as 668.41 ± 12.70 mg GAE/kg fresh weight, EC50 value was 24.28 ± 0.69 mg/ml. The phenolic compounds identified in apple juice by HPLC were catechin, chlorogenic acid, epicatechin and kaempferol. An increase in the amount of phenolic compounds was determined for 20 minutes at 80 °C. When FTIR spectra were examined, it was determined that the specific bands were compatible with epicatechin and chlorogenic acid and these results were parallel to HPLC. In addition, no change in phenolic compounds was observed with FTIR spectroscopy.

Anahtar Kelimeler

• Apple Juice
• Thermal Treatment
• Phenolic Compound
• FTIR
• HPLC

Kaynakça

1. Abbas, O., Compèreb, G., Larondelleb, Y., Pompeuc, D., Rogezd, H., Baetena, V., 2017. Phenolic Compound Explorer: A Mid-Infrared Spectroscopy Database. Vibrational Spectroscopy, 92, 111-118.
2. Abid, M., Jabbar, S., Hu, B., Hashim, M.M., Wu, T., Lei, S., Khan, M.K., Zeng, X., 2014. Thermosonication as a Potential Quality Enhancement Technique of Apple Juice. Ultrasonics Sonochemistry, 21, 984-990.
3. Aguilar-Rosas, S.F., Ballinas-Casarrubias, M.L., Nevarez-Moorillon, G.V., Martin-Belloso, O., Ortega-Rivas, E., 2007. Thermal and Pulsed Electric Fields Pasteurization of Apple Juice: Effects on Physicochemical Properties and Flavour Compounds. Journal of Food Engineering, 83, 41-46.
4. Anonim, 2011, “Türkiye meyve suyu v.b. ürünler sanayi raporu”, https://www.meyed.org.tr (Erişim tarihi: 15 Mart 2015). Bahukhandı, A., Dhyanı, P., Bhatt, I.D., Rawal, R.S., 2018. Variation in Polyphenolics and Antioxidant Activity of traditional Apple Cultivars from West Himalaya, Uttarakhand. Horticultural Plant Journal, 4(4), 151-157.
5. Baltacıoğlu, H., Bayındırlı, A., Severcan, M., Severcan, F., 2015. Effect of Thermal Treatment on Secondary Structure and Conformational Change of Mushroom Polyphenol Oxidase (PPO) as Food Quality Related Enzyme: A FTIR Study. Food Chemistry, 187, 263-269.
6. Başlar, M., Ertugay, M.F., 2013. The Effect of Ultrasound and Photosonication Treatment on Polyphenoloxidase (PPO) Activity, Total Phenolic Component and Colour of Apple Juice. International Journal of Food Science and Technology, 48, 886-892.
7. Blois, M.S., 1958. Antioxidant Determinations by the Use of Stable Free Radical. Nature, 181(4617) 1199-1200.
8. Cemeroğlu, B., 2010. Gıda Analizleri. Gıda Teknolojileri Derneği Yayınları, 34.

9. Cemeroğlu, B., Yemenicioğlu, A., Özkan, M., 2001. Meyve ve Sebzelerin Bileşimi Soğukta Depolanmaları. Gıda Teknolojisi Derneği Yayınları, 24.
10. Ćetković, G., Čanadanović-Brunet, J., Djilas, S., Savatović, S., Mandić, A., Tumbas, V., 2008. Assessment of Polyphenolic Content and in vitro Antiradical Characteristics of Apple Pomace. Food Chemistry, 109(2), 340-347.
11. Dubey, A., Diwakar, S.K., Rawat, S.K., Kumar, P., Batra, N., Joshi, A., Singh, J., 2007. Characterization of Ionically Bound Peroxidases from Apple (Mallus pumilus) Fruits. Preparative Biochemistry and Biotechnology, 37(1), 47-58.
12. Erdoğan, S.S., Demirci, M., 2014. Elmanın Fenolik Bileşen ve Lif İçeriği Atatürk Bahçe Kültürleri Merkez Araştırma Enstitüsü. Gıda Teknolojisi Bölümü, 43(1-2), 41-52.
13. He, Z., Tao, Y., Zeng, M., Zhang, S., Tao, G., Qin, F., Chen, J., 2016. High Pressure Homogenization Processing, Thermal Treatment and Milk Matrix Affect in vitro Bioaccessibility of Phenolics in Apple, Grape and Orange Juice to Different Extents. Food Chemistry, 200, 107-116.
14. Karadeniz, F., Ekşi, A., 2001. Elma Suyunda Fenolik Madde Dağılımı Üzerine Araştırma. Tarım Bilimleri Dergisi, 7(3), 135-141.
15. Karaman, Ş., 2008. Türkiye’de Yetiştirilen Bazı Elma Çeşitlerinin Toplam Antioksidan Kapasitelerinin ve Antioksidan Özellik Gösteren Başlıca Bileşenlerinin Karşılaştırılması. Yüksek Lisans Tezi, İstanbul Üniversitesi Fen Bilimleri Enstitüsü, İstanbul.
16. Krapfenbauer, G., Kinner, M., Gössinger, M., Schönlechner, R., Berghofer, E., 2006. Effect of Thermal Treatment on the Quality of Cloudy Apple Juice. Journal of Agricultural and Food Chemistry, 54(15), 5453-5460.
17. Kuşçu, A., Bulantekin, Ö., 2016. The Effects of Production Methods and Storage on the Chemical Constituents of Apple Pekmez. Journal of Food Science and Technology, 53(7), 3038-3092.
18. Liu, F., Han, Q., Ni, Y., 2017. Comparison of Biochemical Properties and Thermal Inactivation of Membrane-Bound Polyphenol Oxidase from Three Apple Cultivars (Malus domestica Borkh). International Journal of Food Science and Technology, 17, 1-8.
19. Lo Scalzo, R., Iannoccari, T., Summa, C., Morelli, R., Rapisarda, P., 2004. Effect of Thermal Treatments on Antioxidant and Antiradical Activity of Blood Orange Juice. Food Chemistry, 85(1), 41–47.
20. Lu, X., Wang, J., Al-Qadiri, H.M., Ross, C.F., Powers, J.R., Tang, J., Rasco, B.A., 2011. Determination of Total Phenolic Content and Antioxidant Capacity of Onion (Allium cepa) and Shallot (Allium oschaninii) Using Infrared Spectroscopy. Food Chemistry, 129, 637-644.
21. Markowski, J., Płocharski W., 2006. Determination of Phenolic Compounds in Apples and Processed Apple Products. Journal of Fruit and Ornamental Plant Research, 14(2), 133-142.
22. Meral, R., 2016. Farklı Isıl İşlem Uygulamalarının Fenolik Bileşenler Üzerine Etkisi. Yüzüncü Yıl Üniversitesi Fen Bilimleri Enstitüsü Dergisi/ Journal of The Institute of Natural and Applied Sciences, 21(1), 55-67.
23. Nayak, P.K., Rayaguru, K., Krishnan, K.R., 2017. Quality Comparison of Elephant Apple Juices After High-Pressure Processing and Thermal Treatment. Journal Science Food Agricultural 97, 1404-1411.
24. Niu, S., Xu, Z., Fang, Y., Zhang, L., Yang, Y., Liao, X., Hu, X., 2010. Comparative Study on Cloudy Apple Juice Qualities from Apple Slices Treated by High Pressure Carbon Dioxide and Mild Heat. Innovative Food Science and Emerging Technologies, 11(1), 91-97.
25. Okur, İ., Baltacıoğlu C., Baltacıoğlu H., Alpas H., Ağçam E., 2019. Evaluation of the Effect of Different Extraction Techniques on Sour Cherry Pomace Phenolic Content and Antioxidant Activity and Determination of Phenolic Compounds by FTIR and HPLC. Waste and Biomass Valorization.
26. Önez, Z., 2006. Üzümden (Vitis Vinifera L.) İzole Edilen Polifenol Oksidaz Enziminin Özelliklerinin Belirlenmesi. Yüksek Lisans Tezi, Ankara Üniversitesi Fen Bilimleri Enstitüsü, Ankara.
27. Pearson, D.A., Tan, C.H., German, J.B., Davis, P.A., Gershwin, M.E., 1999. Apple Juice Inhibits Human Low Density Lipoprotein Oxidation. Life Sciences, 64(21), 1913-1920.
28. Riener, J., Noci, F. Cronin, D.A., Morgan, D.J., Lyng, J.G., 2008. Combined Effect of Temperature and Pulsed Electric Fields on Apple Juice Peroxidase and Polyphenoloxidase Inactivation. Food Chemistry, 109(2), 402-407.
29. Singleton, V.L., Rossi, J.A., 1965. Colorimetry of Total Phenolics with Phosphomolybdic-Phosphotungstic Acid Reagents. American Journal of Enology and Viticulture, 16, 144-158.
30. Sulaiman, A., Farid, M., Silva, F.V., 2016. Quality Stability and Sensory Attributes of Apple Juice Processed by Thermosonication, Pulsed Electric Field and Thermal Processing. Food Science and Technology International, 23(3), 265-276.
31. Tahir, H.E., Xiaobo, Z., Zhihua, L., Jiyong, S., Zhai, X., Wang, S., Mariod, A.A., 2017. Rapid Prediction of Phenolic Compounds and Antioxidant Activity of Sudanese Honey Using Raman and Fourier Transform Infrared (FT-IR) Spectroscopy. Food Chemistry, 226, 202-211.
32. Terefe, N.S., Buckow, R., Versteeg, C., 2014. Quality-Related Enzymes in Fruit and Vegetable Products: Effects of Novel Food Processing technologies Part 1: High-Pressure Processing. Critical Reviews in Food Science and Nutrition, 54(1), 24-63.
33. Yemenicioğlu, A., Özkan, M., Cemeroğlu, B., 1997. Heat Inactivation Kinetics of Apple Polyphenoloxidase and Activation of its Latent Form. Journal of Food Science, 62(3), 508-510.
34. Yıldırım, M., Benzer, F., Çimen, M., Barış, D., Yıldırım H., Sanyürek, N.K., Karakavuk, E., 2019. Isparta’da Yetişen Bazı Elma Çeşitlerinin Meyve Eti, Kabuk ve Çekirdek Yuvasındaki Antioksidan Kapasitesinin Belirlenmesi. International Journal of Pure and Applied Sciences, 5(1), 31-36.
35. Yi, J., Kebede, B.T., Hai Dang, D.N., Buvé, C., Grauwet, T., Van Loey, A., Hu, X., Hendrickx, M., 2017. Quality Change During High Pressure Processing and Thermal Processing of Cloudy Apple Juice. LWT- Food Science and Technology, 75, 85-92.