Structural insights of two novel N-acetyl-glucosaminidase enzymes through in silico methods

Şahutoğlu, Arif Sercan; Duman, Hatice; Frese, Steven A.; Karav, Sercan

doi:10.3906/kim-2006-19

Cited by 6 publications

(2 citation statements)

References 42 publications

(36 reference statements)

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“…To date, only EndoBI-1 has been shown to be active in the gut of healthy breastfed infants colonized by B. infantis EVC001 (92), but both are likely to be expressed in vivo. Interestingly, EndoBI-1 and EndoBI-2 have different distributions among strains of Bifidobacterium found in infants compared to adults, which may further suggest the importance of these enzymes in healthy gut microbiome formation in both adults and infants (114).…”

Section: Release Of N-glycans From Glycoproteinsmentioning

confidence: 99%

Potential Applications of Endo-β-N-Acetylglucosaminidases From Bifidobacterium longum Subspecies infantis in Designing Value-Added, Next-Generation Infant Formulas

et al. 2021

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Human milk is the optimal source of infant nutrition. Among many other health benefits, human milk can stimulate the development of a Bifidobacterium-rich microbiome through human milk oligosaccharides (HMOs). In recent years, the development of novel formulas has placed particular focus on incorporating some of the beneficial functional properties of human milk. These include adding specific glycans aimed to selectively stimulate the growth of Bifidobacterium. However, the bifidogenicity of human milk remains unparalleled. Dietary N-glycans are carbohydrate structures conjugated to a wide variety of glycoproteins. These glycans have a remarkable structural similarity to HMOs and, when released, show a strong bifidogenic effect. This review discusses the biocatalytic potential of the endo-β-N-acetylglucosaminidase enzyme (EndoBI-1) from Bifidobacterium longum subspecies infantis (B. infantis), in releasing N-glycans inherently present in infant formula as means to increase the bifidogenicity of infant formula. Finally, the potential implications for protein deglycosylation with EndoBI-1 in the development of value added, next-generation formulas are discussed from a technical perspective.

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Section: Release Of N-glycans From Glycoproteinsmentioning

confidence: 99%

Potential Applications of Endo-β-N-Acetylglucosaminidases From Bifidobacterium longum Subspecies infantis in Designing Value-Added, Next-Generation Infant Formulas

et al. 2021

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“…It is also considered as heat resistant and shows activity during 95 °C heat treatments (Karav et al, 2015a;Karav et al, 2015b;Parc et al, 2015). These properties make EndoBI-1 ideal for several applications, especially for the food industry (Karav et al, 2015b;Parc et al, 2015;Şahutoğlu et al, 2020;Duman et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

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

Pekdemir

Duman

Arslan

et al. 2022

Front. Bioeng. Biotechnol.

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Conjugated 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.

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Finding novel enzymes by in silico bioprospecting approach

Kamble

Singh

2022

Value-Addition in Food Products and Processing Through Enzyme Technology

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Structural insights of two novel N-acetyl-glucosaminidase enzymes through in silico methods

Cited by 6 publications

References 42 publications

Potential Applications of Endo-β-N-Acetylglucosaminidases From Bifidobacterium longum Subspecies infantis in Designing Value-Added, Next-Generation Infant Formulas

Potential Applications of Endo-β-N-Acetylglucosaminidases From Bifidobacterium longum Subspecies infantis in Designing Value-Added, Next-Generation Infant Formulas

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

Finding novel enzymes by in silico bioprospecting approach

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