İmmobilize ve Serbest Lakkaz Enziminin Optimum Koşullarının Araştırılması ve Renk Giderme İşleminde Kullanımı

Year 2019, Volume: 11 Issue: 1, 153 - 162, 31.01.2019

Haydar Altınok

https://doi.org/10.29137/umagd.508095

Abstract

Bu çalışmada Tramates versicolor dan elde edilen Laccase
enzimi (L), poli(N-izopropilakrilamit)-kalsiyum aljinat (P(NIPA)-CaAlj) taneciklerine
hapsetme yöntemi ile immobilize edildi. İmmobilize enzimin özellikleri ve enzimatik
renk giderme işlemlerinde kullanımı araştırıldı ve serbest enzim ile
karşılaştırıldı.  Michaelis-Menten sabiti
(Km) ve maksimum reaksiyon hızı (Vmax) değerleri sırasıyla serbest enzim için
1,70x10^-2 mM ve 2,08x10^-3 mM.dak^-1 olarak
bulundu. İmmobilize enzim için Km ve Vmax değerleri de sırasıyla 4,80x10^-2
mM ve 8,70 x 10^-3 mM.dak^-1 olarak bulundu. Optimum
pH değerleri serbest enzim için 5,0 ve immobilize enzim için 6,0 olarak
belirlendi. Optimum sıcaklık sırasıyla serbest lakkaz ve immobilize lakkaz için
40^oC ve 45^oC olarak belirlendi. 4 ^oC tutulan
serbest lakkazın 30 günlük depolama sonrasında orijinal aktivitesinin % 60'ı
koruduğu bulunurken aynı koşullarda tutulan immobilize enzimin ise orijinal
aktivitesinin% 83’ ünü koruduğu bulundu. İmmobilize enzimin 10 kez tekrar
kullanım sonrasında orijinal aktivitesinin % 77'sini koruduğu bulunmuştur. Metil
oranjın renginin giderilmesinde, serbest lakkaz ve immobilize lakkaz için renk
giderme yüzdeleri sırasıyla %73 ve %70 olarak bulundu

Keywords

Lakkaz, immobilizasyon, hapsetme, kalsiyum-aljinat, N-izopropilakrilamit, renk giderme

Investigation of the optimum conditions of immobilized and free laccase enzyme and its use in decolorization process

Abstract

In this study, Laccase enzyme (L) from Tramatesversicolor was entrapped into poly(N-isopropylacryl amide)-calcium alginate (P(NIPA)-CaAlj)  beads The properties of the immobilized enzyme and use of enzymatic decolorization processes were investigated and compared with those of the free enzyme. Michaelis-Menten constant (Km) and maximum reaction rate (Vmax) values were found to be 1.70x10^-2 mM and 2.08x10^-3 mM.min^-1 for free enzyme respectively. Km and Vmax values were found as 4.80x10^-2 mM and 8.70 x 10^-3 mM.min^-1 for entrapped enzymes respectively. Optimum pH was determined as 5.0 and 6.0 and optimum temperature determined as 40^oC and 45^oC for free laccase and entrapped laccase respectively. After 30 days of storage at 4 ^oC free laccase retained 60 % of its original activity. Also after 30 days of storage at 4 ^oC, entrapped enzymes were retained 83 % its original activity. Immobilized enzyme was used repeatedly 10 times, were retained 77 % of its original activities. Percent decolorization of methyl orange by free enyzme and entrapped enyzmes were found to be 73% and 70%, respectively. 

Keywords

Laccase, immobilization, entrapment, calcium-alginate, N-isopropylacyrylamide, decolorization

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