Immobilization of a Bifidobacterial Endo-ss-N-Acetylglucosaminidase to Generate Bioactive Compounds for Food Industry

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dc.authoridKaplan, Merve/0000-0002-3322-0988
dc.authoridKARAV, SERCAN/0000-0003-4056-1673
dc.authoridKaryelioglu, Melda/0000-0003-1918-0964
dc.contributor.authorPekdemir, Burcu
dc.contributor.authorDuman, Hatice
dc.contributor.authorArslan, Aysenur
dc.contributor.authorKaplan, Merve
dc.contributor.authorKaryelioglu, Melda
dc.contributor.authorozer, Tolgahan
dc.contributor.authorKayili, Haci Mehmet
dc.date.accessioned2025-01-27T20:29:07Z
dc.date.available2025-01-27T20:29:07Z
dc.date.issued2022
dc.departmentÇanakkale Onsekiz Mart Üniversitesi
dc.description.abstractConjugated N-glycans are considered next-generation bioactive prebiotic compounds due to their selective stimulation of beneficial microbes. These compounds are glycosidically attached to proteins through N-acetylglucosamines via specific asparagine residue (AsN-X-Ser/Thr). Certain bacteria such as Bifidobacterium longum subspecies infantis (B. infantis) have been shown to be capable of utilizing conjugated N-glycans, owing to their specialized genomic abilities. B. infantis possess a unique enzyme, Endo-ss-N-acetylglucosaminidase (EndoBI-1), which cleaves all types of conjugated N-glycans from glycoproteins. In this study, recombinantly cloned EndoBI-1 enzyme activity was investigated using various immobilization methods: 1) adsorption, 2) entrapment-based alginate immobilization, 3) SulfoLink-, and 4) AminoLink-based covalent bonding immobilization techniques were compared to develop the optimum application of EndoBI-1 to food processes. The yield of enzyme immobilization and the activity of each immobilized enzyme by different approaches were investigated. The N-glycans released from lactoperoxidase (LPO) using different immobilized enzyme forms were characterized using MALDI-TOF mass spectrometry (MS). As expected, regardless of the techniques, the enzyme activity decreased after the immobilization methods. The enzyme activity of adsorption and entrapment-based alginate immobilization was found to be 71.55% +/- 0.6 and 20.32% +/- 3.18, respectively, whereas the activity of AminoLink- and SulfoLink-based covalent bonding immobilization was found to be 58.05 +/- 1.98 and 47.49% +/- 0.30 compared to the free form of the enzyme, respectively. However, extended incubation time recovery achieved activity similar to that of the free form. More importantly, each immobilization method resulted in the same glycan profile containing 11 different N-glycan structures from a model glycoprotein LPO based on MALDI-TOF MS analysis. The glycan data analysis suggests that immobilization of EndoBI-1 is not affecting the enzyme specificity, which enables full glycan release without a limitation. Hence, different immobilization methods investigated in this study can be chosen for effective enzyme immobilization to obtain bioactive glycans. These findings highlight that further optimization of these methods can be a promising approach for future processing scale-up and commercialization of EndoBI-1 and similar enzymes.
dc.description.sponsorshipTUBITAK [117z132]; Evolve Biosystem Inc.
dc.description.sponsorshipFunding This study funded by TUBITAK #117z132 and Evolve Biosystem Inc.
dc.identifier.doi10.3389/fbioe.2022.922423
dc.identifier.issn2296-4185
dc.identifier.pmid35935492
dc.identifier.scopus2-s2.0-85135488933
dc.identifier.scopusqualityQ1
dc.identifier.urihttps://doi.org/10.3389/fbioe.2022.922423
dc.identifier.urihttps://hdl.handle.net/20.500.12428/22831
dc.identifier.volume10
dc.identifier.wosWOS:000836968600001
dc.identifier.wosqualityQ1
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.publisherFrontiers Media Sa
dc.relation.ispartofFrontiers in Bioengineering and Biotechnology
dc.relation.publicationcategoryinfo:eu-repo/semantics/openAccess
dc.rightsinfo:eu-repo/semantics/openAccess
dc.snmzKA_WoS_20250125
dc.subjectB
dc.subjectinfantis
dc.subjectEndo-ss-N-acetylglucosaminidase
dc.subjectN-glycans
dc.subjectimmobilization
dc.subjectbioactive compounds
dc.titleImmobilization of a Bifidobacterial Endo-ss-N-Acetylglucosaminidase to Generate Bioactive Compounds for Food Industry
dc.typeArticle

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