Use of poly HEMA-MAH -Cu2+ Microbeads for α-Amylase Immobilization

Yıl 2014, Cilt: 42 Sayı: 3, 323 - 330, 01.09.2014

Ayşegül Ayla Zübeyde Baysal

Öz

In this article poly 2-hydroxyethyl methacrylate-N-methacryloyl- l -histidin methylester [P HEMA-MAH ] microbeads were prepared by suspension polymerization of 2-hydroxyethyl methacrylate HEMA and N-methacryloyl- l -histidine methyl ester MAH . Some properties of the p HEMA-MAH microbeads were determined by using scanning electron microscopy SEM and swelling tests. Cu2+ ions were chelated on the p HEMA-MAH microbeads, then these microbeads were used in the immobilization of α-amylase in batch system. The maximum α-amylase adsorption capacity of the p HEMA-MAH -Cu2+ beads was found to be 14.88 mg/g at pH 6.0. The optimum temperatures for the immobilized and free enzyme were determined to be 30oC and 25 oC, respectively. Kinetic parameters Km, Vmax of the immobilized enzyme were also determined and compared with those of free enzyme. Reuse studies showed that the immobilized enzyme could reuse six times while retaining 76% of its activity.

Anahtar Kelimeler

α-amylase, P HEMA-MAH, microbead, IMAC

Poli HEMA-MAH -Cu2+ Mikrokürelerin α-Amilaz İmmobilizasyonu için Kullanılması

Öz

S Bu çalışmada poli 2-hidroksietil metakrilat-N-metakriloil- L -histidin metilester [P HEMA-MAH ] mikroküreler süspansiyon polimerizasyon yöntemiyle 2-hidroksietil metakrilat HEMA ve N-metakriloil- L -histidin metil ester MAH ile hazırlandı. P HEMA-MAH mikrokürelerin bazı özellikleri taramalı elektron mikroskobu SEM ve şişme testleriyle belirlendi. P HEMA-MAH mikrokürelere Cu2+ bağlandıktan sonra bu mikroküreler α-amilaz immobilizasyonu için kullanıldı. P HEMA-MAH -Cu2+ mikrokürelerin maksimum α-amilaz adsorpsiyon kapasitesi pH 6.0’da 14.88 mg/g olarak bulundu. İmmobilize ve serbest enzim için optimum sıcaklık sırasıyla 30ºC ve 25ºC olarak belirlendi. İmmobilize enzim için kinetik parametreler de Km, Vmax belirlenerek serbest enzim ile karşılaştırıldı. Tekrarlılık çalışmaları, immobilize enzimin orijinal aktivitesinin %76’sını kaybetmeden altı kez tekrar kulanılabilirliğini gösterdi

Anahtar Kelimeler

α-amilaz, P HEMA-MAH, mikroküre, İMAK

Kaynakça

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