Physicochemical Properties of Invertase Partitioned in an Aqueous Two-Phase System of Poly Ethylene Glycol /Sodium Sulfate

Abstract

Biochemical properties of an enzyme are very important for its applications in various fields. In the present work, we have investigated some parameters affecting to the invertase activity and stability which was partitioned in poly ethylene glycol /sodium sulfate aqueous two-phase system ATPS . The optimum temperature and pH of the invertase were found as 50ºC and pH 5.0, respectively. The enzyme was very stable at the temperature ranged from 25 to 50ºC and also in a pH range of 4.0-7.5. The enzyme was stored at 4ºC and lost about only 47% of its initial activity at the end of two months. Kinetic parameters; Km and Vmax were determined as 4.65 mM and 30 U, respectively. The effect of various effectors on the activity of the invertase was also searched. Especially, Mn+2 ions showed an activator effect for invertase. Invertases are still efficient tools in various biotechnological applications especially in the food and beverage industry. The results obtained in this study indicate that, characteristic properties of invertase partitioned by PEG/Na2SO4 aqueous two-phase system is very suitable for its industrial applications. The relatively wide range of pH and temperature for activity and stability exhibited by invertase could be very interesting for the food processes.

Keywords

• Aqueous two-phase system,PEG/salt systems,Partitioning,Invertase

Poli Etilen Glikol /Sodyum Sülfat Sulu İkili-Faz Sisteminde Ayrılan İnvertazın Fizikokimyasal Özellikleri

Abstract

E nzimlerin biyokimyasal özellikleri çeşitli alanlardaki uygulamaları için oldukça önemlidir. Bu çalşmada, poli etilen glikol /sodyum sülfat sulu ikili-faz sistemi ATPS ile ayrılan invertaz enziminin aktivitesi ve kararlılığına etki eden bazı parametreler incelendi. İnvertaz enziminin optimum pH’sı 5.0 ve optimum sıcaklığı 50oC olarak belirlendi. Enzim 25-50oC sıcaklık ve 4.0-7.5 pH aralığında oldukça kararlıdır. Enzim 4oC’de depolandı ve iki ay sonunda başlangıç aktivitesinin sadece % 47’sini kaybetti. Kinetik parametrelerden K ve Vmax sırasıyla 4.65 mM ve 30 U olarak belirlendi. İnvertaz aktivitesine çeşitli efektörlerin etkisi de incelendi. Özellikle, Mn+2 iyonları invertaz için aktivatör etkisi gösterdi. İnvertazlar özellikle gıda ve içeçek sanayindeki çeşitli biyoteknolojik uygulamalarda hala etkin araçlardır. Bu çalışmadan elde edilen sonuçlar, PEG/Na2SO4 sulu ikili-faz sisteminde ayrılan invertazın karakteristik özelliklerinin endüstriyel uygulamaları için oldukça uygun olduğunu gösterdi. İnvertaz aktivitesi ve kararlığı için sıcaklık ve pH’nın geniş aralıkta değişmesi özellikle gıda prosesleri için ilgi çekici olacaktır

Keywords

• Sulu ikilifaz sistemi,PEG/tuz sistemi,Ayrılma,invertaz

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