Investigation of the Polyphenol Oxidase Enzyme Thin Film as a Copper Sulfate (CuSO4) Sensor

Abstract

In this study, thin films were produced with the polyphenol oxidase (PPO) enzyme on chitosan-gelatine bio-composite via the spin coating technique. Bio-composite material was used for the PPO enzyme layer and for stable thin film transfer on the substrate. In the thin films produced, the PPO enzyme layer forms the active site and is the part where the thin film is in direct contact with the external environment. In this way, any external environmental change to the thin film directly affects the PPO enzyme layer. A copper sulfate (CuSO4) solution in pure water solvent was used to observe the external changes in the environment of the PPO enzyme layer. The copper sulfate in this solution is a pollutant and is the harmful chemical to be detected. Therefore, its usability in sensor applications was investigated by measuring the response of the PPO enzyme to copper sulfate. Production stages of thin films were followed by electrical and Quartz crystal microbalance (QCM) methods. In addition, electrical measurements and the QCM system have been used for liquid media sensor measurements of thin films. By recording the time-dependent contaminant response of the PPO enzyme thin film, the usability of enzymes as an active layer in sensor applications was studied.

Keywords

• Polyphenol oxidase enzyme
• bio-sensor
• quartz crystal microbalance

Polifenol Oksidaz Enzim İnce Filminin Bakır Sülfat (CuSO4) Sensörü Olarak Kullanılabilirliğinin İncelenmesi

Abstract

Bu çalışmada spin kaplama tekniği kullanılarak çitosan-jelatin biyokompozit alt-tabakası üzerine polifenol oksidaz (PPO) enzimi ile ince filmler üretilmiştir. Biyo-kompozit malzeme, PPO enzim tabakasının alttaş üzerine kararlı ince film transferi amacıyla kullanılmıştır. Üretilen ince filmlerde PPO enzim tabakası aktif bölgeyi oluşturarak, ince filmin dış ortam ile direkt temas halinde olduğu kısımdır. Bu sayede ince filmin herhangi bir dış ortam değişimi, PPO enzim tabakası üzerine doğrudan etki etmektedir. PPO enzim tabakasının dış ortam değişimlerini duyarlılığını gözlemlemek amacıyla saf su çözücüsünde bakır sülfat (CuSO4) çözeltisi kullanılmıştır. Bu çözeltideki bakır sülfat kirletici olup tespit edilmek istenilen zararlı kimyasal görevindedir. Dolayısıyla, PPO enziminin bakır sülfata karşı tepkisi ölçülerek sensör uygulamalarındaki kullanılabilirliği araştırılmıştır. İnce filmlerin üretim aşamaları elektriksel ve Kuartz kristal mikrobalans (QCM) yöntemleriyle takip edilmiştir. Ayrıca elektriksel ölçümler ve QCM sistemi, ince filmlerin sıvı ortam sensör ölçümleri için kullanılmıştır. PPO enzim ince filminin zamana bağlı kirletici tepkisi kaydedilerek enzimlerin sensör uygulamalarında aktif tabaka olarak kullanılabilirlikleri araştırılmıştır.

Keywords

• Polifenol oksidaz enzimi
• biyo-sensör
• kuartz kristal mikrobalans

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