Impact of orientation of carbohydrate binding modules family 22 and 6 on the catalytic activity of Thermotoga maritima xylanase XynB

Tajwar, Razia; Shahid, Saleem Ullah; Zafar, Rehan; Akhtar, Muhammad

doi:10.1016/j.enzmictec.2017.07.004

Cited by 19 publications

(3 citation statements)

References 41 publications

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“…Our results suggest that the mechanism of EndoS2 relies on an interplay between its GH domain and CBM. Many endoglycosidases have CBMs appended to them; however, they are usually appended in seemingly random orientations relative to the GH domain, and only a few studies have investigated how the orientation of the CBM affects the activity of the GH domain. − It is tempting to speculate that the orientation of the CBM affects EndoS2 activity for several reasons. First, the GH and CBM are located on opposite tips of the “V-shaped” enzyme, with their glycan-binding surfaces pointing inward.…”

Section: Discussionmentioning

confidence: 99%

Molecular Basis of Broad Spectrum N-Glycan Specificity and Processing of Therapeutic IgG Monoclonal Antibodies by Endoglycosidase S2

et al. 2019

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Immunoglobulin G (IgG) glycosylation critically modulates antibody effector functions. Streptococcus pyogenes secretes a unique endo-β- N -acetylglucosaminidase, EndoS2, which deglycosylates the conserved N -linked glycan at Asn297 on IgG Fc to eliminate its effector functions and evade the immune system. EndoS2 and specific point mutants have been used to chemoenzymatically synthesize antibodies with customizable glycosylation for gain of functions. EndoS2 is useful in these schemes because it accommodates a broad range of N -glycans, including high-mannose, complex, and hybrid types; however, its mechanism of substrate recognition is poorly understood. We present crystal structures of EndoS2 alone and bound to complex and high-mannose glycans; the broad N -glycan specificity is governed by critical loops that shape the binding site of EndoS2. Furthermore, hydrolytic experiments, domain-swap chimeras, and hydrogen–deuterium exchange mass spectrometry reveal the importance of the carbohydrate-binding module in the mechanism of IgG recognition by EndoS2, providing insights into engineering enzymes to catalyze customizable glycosylation reactions.

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Section: Discussionmentioning

confidence: 99%

Molecular Basis of Broad Spectrum N-Glycan Specificity and Processing of Therapeutic IgG Monoclonal Antibodies by Endoglycosidase S2

et al. 2019

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“…The designed linker peptide will interfere with the thermostability [ 27 , 28 , 29 ] and enzymatic activity [ 17 , 27 ] of adjacent domains. As previously reported, the flexibility of the linker region is of great significance for enzyme activity [ 30 , 31 , 32 ], so we used a natural 22-residue linker peptide (PEVLPPLPKESRISEGEAVVVG) to connect these domains. The linker we selected contains four Pro and two Ser residues, which can form flexible hinge regions and aid in creating an active spatial conformation between different domains [ 1 , 25 ].…”

Section: Discussionmentioning

confidence: 99%

Improving the Thermostability of a Fungal GH11 Xylanase via Fusion of a Submodule (C2) from Hyperthermophilic CBM9_1-2

Miao

Ying-jie

Zhe

et al. 2021

IJMS

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Xylanases have been applied in many industrial fields. To improve the activity and thermostability of the xylanase CDBFV from Neocallimastix patriciarum (GenBank accession no. KP691331), submodule C2 from hyperthermophilic CBM9_1-2 was inserted into the N- and/or C-terminal regions of the CDBFV protein (producing C2-CDBFV, CDBFV-C2, and C2-CDBFV-C2) by genetic engineering. CDBFV and the hybrid proteins were successfully expressed in Escherichia coli BL21 (DE3). Enzymatic property analysis indicates that the C2 submodule had a significant effect on enhancing the thermostability of the CDBFV. At the optimal temperature (60.0 °C), the half-lives of the three chimeras C2-CDBFV, CDBFV-C2, and C2-CDBFV-C2 are 1.5 times (37.5 min), 4.9 times (122.2 min), and 3.8 times (93.1 min) longer than that of wild-type CDBFV (24.8 min), respectively. More importantly, structural analysis and molecular dynamics (MD) simulation revealed that the improved thermal stability of the chimera CDBFV-C2 was on account of the formation of four relatively stable additional hydrogen bonds (S42-S462, T59-E277, S41-K463, and S44-G371), which increased the protein structure’s stability. The thermostability characteristics of CDBFV-C2 make it a viable enzyme for industrial applications.

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“…However, it has been reported that fusion with CBMs may have different effects on different enzymes and the effects may also vary with different CBMs because of their divergent structures and properties. For instance, an increase in activity of Thermotoga maritima xylanase XynB was reported with the addition of CBM6 and CBM22 (Tajwar et al, 2017). In contrast, the addition of CBM6 to xylanase XynZ from Clostridium thermocellum resulted in decreased activity (Khan et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Expansin assisted bio-affinity immobilization of endoxylanase from Bacillus subtilis onto corncob residue: Characterization and efficient production of xylooligosaccharides

Chang

et al. 2019

Food Chemistry

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A one-step method to immobilize xylanase onto cellulosic material by fusion of expansin from Bacillus subtilis to xylanase LC9 without the requirement of prior purification of enzyme has been developed. Fusion enzyme EXLX-R2-XYN was specifically adsorbed onto corncob residue with high loading capacity due to bioaffinity adsorption of expansin onto cellulose. The immobilization yield was close to 100%, with a recovered activity of 82.4%. The immobilized EXLX-R2-XYN retained 45.3% of its activity after incubation at 70 °C for 3 h, whereas only 16.3% of the activity was left in free form under the same conditions. The conversion yield of XOS by using immobilized EXLX-R2-XYN reached up to 515 mg/g xylan from 2% corncob extracted xylan, which was higher than that of the free enzyme. The hydrolysis products were mainly xylobiose (57.5%) and xylotriose (38.4%), without undesirable xylose production. After five cycles of hydrolysis, more than 70% of conversion was obtained.

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Impact of orientation of carbohydrate binding modules family 22 and 6 on the catalytic activity of Thermotoga maritima xylanase XynB

Cited by 19 publications

References 41 publications

Molecular Basis of Broad Spectrum N-Glycan Specificity and Processing of Therapeutic IgG Monoclonal Antibodies by Endoglycosidase S2

Molecular Basis of Broad Spectrum N-Glycan Specificity and Processing of Therapeutic IgG Monoclonal Antibodies by Endoglycosidase S2

Improving the Thermostability of a Fungal GH11 Xylanase via Fusion of a Submodule (C2) from Hyperthermophilic CBM9_1-2

Expansin assisted bio-affinity immobilization of endoxylanase from Bacillus subtilis onto corncob residue: Characterization and efficient production of xylooligosaccharides

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