Termofilik Anoxybacillus gonensis glukoz izomerazının DEAE-Sefaroz üzerine iyonik ve kovalent immobilizasyonu

Züleyha Akpınar; Merve Kızaklı Yıldırım; Hakan Karaoğlu

doi:10.17714/gumusfenbil.1028883

TR EN

Termofilik Anoxybacillus gonensis glukoz izomerazının DEAE-Sefaroz üzerine iyonik ve kovalent immobilizasyonu

Öz

Bir şekerin diğerine (glukozun fruktoza) dönüştürülmesiyle üretilen yüksek fruktozlu mısır şurubu (HFCS), pazarlama değerine sahiptir. Bu nedenle günümüze kadar farklı kaynaklardan (makro ve mikroorganizmalar) izole edilen farklı glukoz izomerazlar [(GI) (D-ksiloz ketol izomeraz, EC 5.3.1.5)] araştırılmıştır. Ayrıca endüstriyel uygulamalar için GI üretiminin maliyetinin düşürülmesi araştırılmış ve bunun için farklı teknikler uygulanmıştır. Enzim immobilizasyon yaklaşımları, enzimlerin tekrar tekrar kullanılmasına izin verdiği için öne çıkan özelliklere sahiptir. Bu çalışmada Anoxybacillus gönensis G2T (AgoGI) yabani tip enzimlerinin DEAE-sefaroz matriksi üzerinde immobilizasyonu (iyonik ve kovalent) gerçekleştirildi. Çalışmanın bir sonraki aşamasında elde edilen enzimlerin kinetik ve biyokimyasal özellikleri belirlendi. İmmobilize enzimler için optimum sıcaklık ve pH değerleri sırasıyla 85 °C ve 6.50 olarak belirlendi. DEAE-sefaroz üzerinde iyonik bağlı AgoGI'nin kinetik verileri (Vmax ve Km) 4.85±2.09 μmol/dk/mg protein ve 130,57±5,42 mM, aynı matris üzerinde kovalent immobilize AgoGI 40.51± 0.81 μmol/dk/mg protein ve 127,28±2,96 mM’dır. Sonuç olarak, DEAE-sefarozun hem kovalent hem iyonik immobilizasyon için immobilizasyon matriksi olarak kullanılması, glukoz izomerazın biyokimyasal ve kinetik parametreleri üzerinde herhangi bir olumsuz etki göstermedi. Bu nedenle, DEAE-sefaroz üzerinde immobilize edilmiş AgoGI HFCS üretimi için mükemmel ve umut verici bir araçtır.

Anahtar Kelimeler

Anoxybacillus gönensis , DEAE-sefaroz , Glukoz izomeraz , HFCS , İmmobilizasyon

Ionic and covalent immobilization of glucose isomerase of thermophilic Anoxybacillus gonensis on DEAE-sepharose

Abstract

High fructose corn syrup (HFCS), which is produced by the conversion of one sugar into another (glucose to fructose), has a marketing value. Hence, different glucose isomerases [(GI) (D-xylose ketol isomerase, EC 5.3.1.5)] isolated from different sources (macro-and microorganisms) were researched until today. In addition, the cost reduction of GI production for industrial applications has been investigated and applied with different techniques. Enzyme immobilization approaches have prominent features because they allow enzymes to be used repeatedly. In the current study, Anoxybacillus gonensis G2T glucose isomerase (AgoGI) (wild type) were immobilized with ionic and covalent binding on DEAE-sepharose matrix. Afterward, kinetic and biochemical parameters of the immobilized enzymes were evaluated. The pH and temperature parameters, in which the ionic and covalent immobilized enzymes showed the best activity, were determined as 6.50 and 85 °C, respectively. The kinetic data (Vmax and Km) of ionic bound AgoGI on DEAE-sepharose were 4.85±2.09 μmol/min/mg protein and 130,57±5,42 mM, as covalent immobilized AgoGI on the same matrix were 40.51± 0.81 μmol/min/mg protein µmol/min and 127,28±2,96 mM, respectively. Consequently, the usage of DEAE-sepharose for both covalent and ionic immobilization as immobilization matrix did not exhibit any negative effects on biochemical and kinetic parameters of glucose isomerase. Therefore, immobilized AgoGI on DEAE-sepharose was an excellent and promising tool for HFCS production.

Keywords

Anoxybacillus gonensis , DEAE-sepharose , Glucose isomerase , HFCS , Immobilization

Kaynakça

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Ayrıntılar

Birincil Dil

Türkçe

Konular

Mühendislik

Bölüm

Araştırma Makalesi

Yazarlar

Züleyha Akpınar
0000-0003-0102-6651
Türkiye

Merve Kızaklı Yıldırım
0000-0002-2040-3881
Türkiye

Hakan Karaoğlu ^*
0000-0003-4615-1157
Türkiye

Yayımlanma Tarihi

15 Temmuz 2022

Gönderilme Tarihi

26 Kasım 2021

Kabul Tarihi

19 Nisan 2022

Yayımlandığı Sayı

Yıl 2022 Cilt: 12 Sayı: 3

DOI

https://doi.org/10.17714/gumusfenbil.1028883

IZ

https://izlik.org/JA68NT82MT

APA

Akpınar, Z., Kızaklı Yıldırım, M., & Karaoğlu, H. (2022). Termofilik Anoxybacillus gonensis glukoz izomerazının DEAE-Sefaroz üzerine iyonik ve kovalent immobilizasyonu. Gümüşhane Üniversitesi Fen Bilimleri Dergisi, 12(3), 793-802. https://doi.org/10.17714/gumusfenbil.1028883

Cited By

The use of immobilized enzyme in starch bioconversion: An update review

BIO Web of Conferences

https://doi.org/10.1051/bioconf/20249601028

Xylose isomerase: From fundamental research to applied enzyme technology

Journal of Biotechnology

https://doi.org/10.1016/j.jbiotec.2025.04.002

e-ISSN: 2146-538X Yayın Modeli: Sürekli Yayın Atıf Dizinleri: TR Dizin , SCOPUS

Termofilik Anoxybacillus gonensis glukoz izomerazının DEAE-Sefaroz üzerine iyonik ve kovalent immobilizasyonu

Öz

Anahtar Kelimeler

Ionic and covalent immobilization of glucose isomerase of thermophilic Anoxybacillus gonensis on DEAE-sepharose

Abstract

Keywords

Kaynakça

Ayrıntılar

Birincil Dil

Konular

Bölüm

Yazarlar

Yayımlanma Tarihi

Gönderilme Tarihi

Kabul Tarihi

Yayımlandığı Sayı

DOI

IZ

Kaynak Göster

Cited By

The use of immobilized enzyme in starch bioconversion: An update review

Xylose isomerase: From fundamental research to applied enzyme technology