Mantar Lakkazı’nın alginat jellerde tutuklanması ve atık sularda boyar madde giderilmesinde kullanılmas
Yükleniyor...
Dosyalar
Tarih
2013
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Trakya Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışma kapsamında, Boletus edulis mantarından izole edilen lakkaz enzimi alginat/karragenan boncuklara immobilize edildi, serbest ve immobilize enzimlerin bazı özellikleri belirlendi ve bazı tekstil boyaları kullanılarak boya giderme aktiviteleri incelendi. Boletus edulis mantarından izole edilerek % 40-60 (NH4)2SO4 tuz çöktürmesi ve diyaliz ile elde edilen lakkaz enzimi alginat/karragenan boncuklarda tutuklandı. Lakkaz aktivitesi tayini substrat olarak 2,2'-azino-bis(3-etilbenzotiyazolin-6-sülfonik asit) (ABTS) kullanılarak gerçekleştirildi. Protein tayini Bradford yöntemi ile yapıldı. Serbest ve immobilize enzimin lakkaz aktiviteleri sırasıyla; 281.896 U/mg ve 41.818 U/mg olarak belirlendi. İmmobilizasyon ve çalışma koşullarının optimizasyon çalışması sonucunda; damlatma çözeltisi olarak % 2’lik CuCl2, Destek:Yüklenen enzim miktarı oranı 1:5 (w:v), boncuk boyutu 1.8 mm, aktivite tayininde kullanılacak boncuk miktarı 0.25 g olarak belirlendi. Serbest enzim için optimum pH 2.0, optimum sıcaklık 25 °C; Km ve Vmax değerleri sırasıyla 0.333 mM ve 196.078 U/mg olarak belirlendi. İmmobilize enzim için ise optimum pH 3.5, optimum sıcaklık 37 °C, Km ve Vmax değerleri sırasıyla 0.642 mM ve 33.445 U/mg olarak belirlendi. Tekrar kullanılabilirlik çalışmasında immobilize enzimin 14. döngü sonunda aktivitesinin % 69’unu koruduğu gözlendi. Depo kararlılığı çalışmasında, +4 °C’de, serbest enzimin 28 günün sonunda başlangıç aktivitesini % 77 oranında, immobilize enzimin ise yaklaşık % 92 oranında koruduğu gözlendi. Termal kararlılık çalışmasında, serbest enzim 70 °C’de aktivitesini büyük ölçüde korurken, immobilize enzimin 70 °C’de aktivitesinin yaklaşık % 55’ini kaybettiği gözlendi. Renk giderme çalışmasında, Reactive Blue 19, Reactive Red 239, Reactive Orange ve Reactive Violet boyar maddeleriyle serbest enzim kullanılarak % 85-90 arasında renk giderme aktivitesi elde edildi. En yüksek renk giderme aktivitesi,% 88.9 ile Reactive Orange boyar maddesinde belirlendi. Anahtar Kelimeler : Lakkaz, Boletus edulis, immobilizasyon, alginat, karragenan, renk giderme aktivitesi
Abstract
In this study, laccase isolated from Boletus edulis mushroom was immobilized alginate+carrageenan gel beads, some characteristics of free and immobilized laccases were determined, and their decolorisation activities evaluated by using same textile dyes. Laccase isolated from fresh Boletus edulis mushroom was applied 40-60 % (NH4)2SO4 precipitation and dialysis, and then was immobilized alginate+carrageenan gel beads. Laccase activity was carried out by using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonicacid) (ABTS) as substrate. Protein determinations were done with Bradford method. Laccase activities were determined as 281.896 U/mg and 41.818 U/mg for free and immobilized enzyme, respectively. Dropping solution was determined as 2 % CuCl2. Support:Loading enzyme ratio was 1:5 (w:v). Optimum bead diameter and optimum bead amount were determined to be 1.8 mm and 0.25 g, respectively. For free laccase, optimum pH was 2.0, optimum temperature 25 °C, Km and Vmax values were 0.333 mM and 196.078 U/mg protein, respectively. For immobilized laccase, these parameters were pH 3.5, 37 °C, 0.642 mM and 33.445 U/mg, respectively. In reusability assay, immobilized laccase saved 69 % of its initial activity at the end of 14 cycles. In the storage stability assay at 4 °C, free and immobilized enzyme saved 77 % and about 92 % of their initial activities at the end of 28 days, respectively. In thermal stability assay, free enzyme saved largely its activity at 70 °Cwhile immobilized enzyme lost 55 % of its initial activity. In decoulorisation assay, using Reactive Blue 19, Reactive Red 239, Reactive Orange and Reactive Violet dyes, 85-90 % decolorization was obtained for the free enzyme. The most effective decolorization activity was determined to be 88.9 % with Reactive Orange dye. Keywords : Laccase, Boletus edulis, immobilization, alginate, carrageenan, decolorization activity
Abstract
In this study, laccase isolated from Boletus edulis mushroom was immobilized alginate+carrageenan gel beads, some characteristics of free and immobilized laccases were determined, and their decolorisation activities evaluated by using same textile dyes. Laccase isolated from fresh Boletus edulis mushroom was applied 40-60 % (NH4)2SO4 precipitation and dialysis, and then was immobilized alginate+carrageenan gel beads. Laccase activity was carried out by using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonicacid) (ABTS) as substrate. Protein determinations were done with Bradford method. Laccase activities were determined as 281.896 U/mg and 41.818 U/mg for free and immobilized enzyme, respectively. Dropping solution was determined as 2 % CuCl2. Support:Loading enzyme ratio was 1:5 (w:v). Optimum bead diameter and optimum bead amount were determined to be 1.8 mm and 0.25 g, respectively. For free laccase, optimum pH was 2.0, optimum temperature 25 °C, Km and Vmax values were 0.333 mM and 196.078 U/mg protein, respectively. For immobilized laccase, these parameters were pH 3.5, 37 °C, 0.642 mM and 33.445 U/mg, respectively. In reusability assay, immobilized laccase saved 69 % of its initial activity at the end of 14 cycles. In the storage stability assay at 4 °C, free and immobilized enzyme saved 77 % and about 92 % of their initial activities at the end of 28 days, respectively. In thermal stability assay, free enzyme saved largely its activity at 70 °Cwhile immobilized enzyme lost 55 % of its initial activity. In decoulorisation assay, using Reactive Blue 19, Reactive Red 239, Reactive Orange and Reactive Violet dyes, 85-90 % decolorization was obtained for the free enzyme. The most effective decolorization activity was determined to be 88.9 % with Reactive Orange dye. Keywords : Laccase, Boletus edulis, immobilization, alginate, carrageenan, decolorization activity
Açıklama
Yüksek Lisans Tezi
Anahtar Kelimeler
Lakkaz, Boletus Eduli, İmmobilizasyon, Algina, karragenaN, Renk Giderme Aktivitesi, Laccase, Boletus Edulis, İmmobilization, Alginate, Carrageena, Decolorization Activity
