Yıl 2018, Cilt 21, Sayı 3, Sayfalar 286 - 291 2018-06-15

Understanding of the Contribution of Fetuin O-glycans for the Release of New Bioactive Compounds by a Novel Endo-β-N-acetylglucosaminidase
Fetuin O-glikanlarının, Bioaktif N-Glikanların Yeni Endo-B-N-asetilglukozaminidaz Tarafından İzole Edilmesindeki Katkısının Belirlenmesi

Sercan KARAV [1]

44 115

Bovine fetuin is a model protein to study the activity of various glycosidases since it contains both N- and O- glycans attached to the polypeptide chain. We recently showed a novel glycosidase, endo-β-N-acetylglucosaminidase isolated from an infant gut microbe, Bifidobacterium infantis. This enzyme is capable of cleaving the N-N’-diacetyl chitobiose moiety found in the N-glycan core of a wide variety of proteins. It is considered a promising approach to release N-glycans from complex substrates such as whey proteins due to its high activity and wide substrate specificity. Moreover, it also maintains its activity at high temperatures enabling the use of this enzyme in thermal dairy processes such as during the pasteurization. Bovine whey is a potential source of glycans providing million tons of glycoproteins annually. Application of EndoBI-1 on bovine whey is challenging due to the complexity of the whey proteins and their O-glycosylation pattern. O-glycans are considered to be a protective agent for N-deglycosylation that hinders the isolation of these recently found novel compounds. In this study, O-glycans were removed from fetuin (both O- and N- glycosylated model glycoprotein) and the contribution of O-glycans to the accessibility of EndoBI-1 to bovine fetuin N-glycans were tested. Released glycans were characterized by advanced mass spectrometry and 22 different N-glycans (including isomers) were monitored. According to the results, it was shown that removing O-glycans from Fetuin increases the Kcat/Km value 0.52 to 1.54 ml/mg x min-1and the affinity of EndoBI-1 (Km value from 0.32 to 0.22 mg/ml) to target N-glycans enabling more feasible application of this enzyme in dairy streams.

İnek fetuini N- ve O- glikanları içerdiğinden dolayı farklı glikosidazların aktivitesini test etmek için kullanılan model bir proteindir. Yeni bir glikosidaz olan endo-B-N-asetilglukozaminidaz enzimi (EndoBI-1) bebek bağırsaklarında bulunan Bifidobakteri infantis’ten tarafımızca daha önceden izole edilmiştir. Bu enzim farklı protein yapılarında bulunan N-glikan merkezlerini kesebilmektedir. Enzimin yüksek aktivite ve geniş substrat aralığından dolayı, peynir altı suyu proteinleri gibi karmaşık yapılarda aktivite gösterebilmektedir. Ayrıca, bu enzim aktivitesini yüksek sıcaklıklarda koruyabildiği için, pastörizasyon gibi ısıl işlem gerektiren süreçlerde de kullanılabilmektedir. İnek peynir altı suyu, yıllık milyonlarca ton üretilen bir glikoprotein kaynağıdır. Fakat, EndobI-1 enziminin bu substrata uygulanması, peynir altı proteinlerinin O-glikanları tarafından bloke edilip aktiviteyi düşürdüğü düşünülmemektedir. O-glikanların, proteinleri bir  korucuyu görevi ile koruyarak, yeni bir prebiyotik kaynağı olarak kabul edilen N-glikanların izolasyonunu zorlaştırmakta olduğu sanılmaktadır. Bu çalışmada, O-glikanlar fetuinden ayrılarak, bunun N-glikanların enzimatik olarak EndoBI-1 tarafından ayrılmasına olan katkısı incelenmiştir. Ayrılan glikanlar kütle spektrometresi ile analiz edilmiş ve izomerler dahil 22 farklı yapı gözlemlenmiştir. Sonuçlara göre, O-glikanların fetuinden ayrılması, N-glikan izolasyon etkinliğini (Kcat/Km) 0.52 ‘den 1.54 ml/mg x min-1’a çıkarırken, Km değerini 0.32 ‘den 0.22 mg/ml’ye düşürmüştür. Bu sonuçlara göre, EndobI-1 süt endüstrisinde çok daha etkili bir şekilde kullanabilmesinin yolu açılmıştır.

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Orcid: 0000-0003-4056-1673
Yazar: Sercan KARAV (Sorumlu Yazar)

Bibtex @araştırma makalesi { ksutarimdoga428544, journal = {Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi}, issn = {}, eissn = {2619-9149}, address = {Kahramanmaraş Sütçü İmam Üniversitesi}, year = {2018}, volume = {21}, pages = {286 - 291}, doi = {10.18016/ksudobil.335396}, title = {Understanding of the Contribution of Fetuin O-glycans for the Release of New Bioactive Compounds by a Novel Endo-β-N-acetylglucosaminidase}, key = {cite}, author = {KARAV, Sercan} }
APA KARAV, S . (2018). Understanding of the Contribution of Fetuin O-glycans for the Release of New Bioactive Compounds by a Novel Endo-β-N-acetylglucosaminidase. Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 21 (3), 286-291. DOI: 10.18016/ksudobil.335396
MLA KARAV, S . "Understanding of the Contribution of Fetuin O-glycans for the Release of New Bioactive Compounds by a Novel Endo-β-N-acetylglucosaminidase". Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi 21 (2018): 286-291 <http://dogadergi.ksu.edu.tr/issue/37038/428544>
Chicago KARAV, S . "Understanding of the Contribution of Fetuin O-glycans for the Release of New Bioactive Compounds by a Novel Endo-β-N-acetylglucosaminidase". Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi 21 (2018): 286-291
RIS TY - JOUR T1 - Understanding of the Contribution of Fetuin O-glycans for the Release of New Bioactive Compounds by a Novel Endo-β-N-acetylglucosaminidase AU - Sercan KARAV Y1 - 2018 PY - 2018 N1 - doi: 10.18016/ksudobil.335396 DO - 10.18016/ksudobil.335396 T2 - Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi JF - Journal JO - JOR SP - 286 EP - 291 VL - 21 IS - 3 SN - -2619-9149 M3 - doi: 10.18016/ksudobil.335396 UR - http://dx.doi.org/10.18016/ksudobil.335396 Y2 - 2017 ER -
EndNote %0 Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi Understanding of the Contribution of Fetuin O-glycans for the Release of New Bioactive Compounds by a Novel Endo-β-N-acetylglucosaminidase %A Sercan KARAV %T Understanding of the Contribution of Fetuin O-glycans for the Release of New Bioactive Compounds by a Novel Endo-β-N-acetylglucosaminidase %D 2018 %J Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi %P -2619-9149 %V 21 %N 3 %R doi: 10.18016/ksudobil.335396 %U 10.18016/ksudobil.335396
ISNAD KARAV, Sercan . "Fetuin O-glikanlarının, Bioaktif N-Glikanların Yeni Endo-B-N-asetilglukozaminidaz Tarafından İzole Edilmesindeki Katkısının Belirlenmesi". Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi 21 / 3 (Haziran 2018): 286-291. http://dx.doi.org/10.18016/ksudobil.335396