Amin fonksiyonel silika nanotaneciklere kovalent bağlanma ile lakkaz enzim immobilizasyonu ve enzimatik renk giderimi
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Tarih
2019
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Kırıkkale Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
Bu çalışmada lakkaz enziminin karbodimit (EDC) ve hidroksisüksinimid (NHS) ile aktifleştirilmiş amin fonksiyonel silika nanotaneciklere kovalent bağlanma yöntemi ile immobilizasyonu gerçekleştirildi. İmmobilize enzimlerin özellikleri ve enzimatik renk giderimi araştırıldı ve serbest lakkaz enzim ile karşılaştırıldı. Michaelis-Menten sabiti (Km) ve maksimum reaksiyon hızı (Vmax) değerleri sırasıyla serbest enzim için 5,14 mM ve 1,217 mM.dakika-1 olarak bulundu. EDC ile aktifleştirilmiş immobilize enzim için Km ve Vmax değerleri de sırasıyla 21,28 mM ve 0,611 mM.dakika-1, NHS ile aktifleştirilmiş immobilize enzim için 21,12 mM ve 0,686 mM.dakika-1 olarak bulundu. Optimum pH değerleri serbest lakkaz için 5,0 ve immobilize enzimler için 6,0 olarak bulundu. Optimum sıcaklık serbest lakkaz ve immobilize enzimler için 40 oC olarak bulundu. + 4 °C' da 60 günlük depolama süresinin sonrasında serbest enzimin bağıl aktivitesindeki düşüş % 93 iken, EDC ve NHS ile immobilize enzimlerin aktivitelerindeki düşüş ise sırasıyla % 58 ve % 62,5 idi. EDC ve NHS kullanılarak hazırlanan immobilize enzimlerin 15 kez tekrar kullanım sonrasında orijinal aktivitelerini sırasıyla % 89 ve % 87 koruduğu gözlenmiştir. Metil oranjın renginin giderilmesinde kullanılan serbest ve EDC ve NHS ile immobilize enzimlerin renk giderme yüzdeleri sırasıyla % 59,4, % 69, % 67 olarak bulundu.
In this study, the laccase enzyme was immobilized covalent bonding method to amin funtionalized silica nanoparticles preactivated with carbodimide (EDC) and hydroxysuccinimide (NHS). Properties of immobilized enzyme and enzymatic discoloration were investigated and compared with free laccase enzyme. The Michaelis-Mentenconstant (Km) and maximum reaction rate (Vmax) values were found to be 5.14 Mm and 1.217 mM.min-1 for the free enzyme, respectively. Km and Vmax values were of 21.28 mM and 0.611 mM.min-1, for immobilized enzyme with preactivated EDC, 21.12 mM and 0.686 mM.min-1, for immobilized enzyme with preactivated NHS, respectively. Optimum pH values were determined as 5,0 for free laccase and 6,0 for immobilized enzymes. Optimum temperature determined as 40 oC for free laccase and immobilized enzymes, respectively. After 60-days of storage period at + 4 oC, while the decreasing of the activity of free enzyme was of 93 %, the decreasing of the activities of immobilized enzyme with EDC and NHS were of were 58 % and 62,5 %, respectively. It was observed that after 15 reusing of the immobilized enzyme susing EDC and NHS, which retained their original activities of 89 % and 87 %, respectively. The decolorization percentages of the free and immobilized enzymes with EDC and NHS, which were used to decolorize the methyl orange, were determined as 59,4 %, 69 % and 67 %, respectively.
In this study, the laccase enzyme was immobilized covalent bonding method to amin funtionalized silica nanoparticles preactivated with carbodimide (EDC) and hydroxysuccinimide (NHS). Properties of immobilized enzyme and enzymatic discoloration were investigated and compared with free laccase enzyme. The Michaelis-Mentenconstant (Km) and maximum reaction rate (Vmax) values were found to be 5.14 Mm and 1.217 mM.min-1 for the free enzyme, respectively. Km and Vmax values were of 21.28 mM and 0.611 mM.min-1, for immobilized enzyme with preactivated EDC, 21.12 mM and 0.686 mM.min-1, for immobilized enzyme with preactivated NHS, respectively. Optimum pH values were determined as 5,0 for free laccase and 6,0 for immobilized enzymes. Optimum temperature determined as 40 oC for free laccase and immobilized enzymes, respectively. After 60-days of storage period at + 4 oC, while the decreasing of the activity of free enzyme was of 93 %, the decreasing of the activities of immobilized enzyme with EDC and NHS were of were 58 % and 62,5 %, respectively. It was observed that after 15 reusing of the immobilized enzyme susing EDC and NHS, which retained their original activities of 89 % and 87 %, respectively. The decolorization percentages of the free and immobilized enzymes with EDC and NHS, which were used to decolorize the methyl orange, were determined as 59,4 %, 69 % and 67 %, respectively.
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Kimya, Chemistry, , , , , , , , , ,
