Selective Oxidation of <i>N</i>-Glycolylneuraminic Acid Using an Engineered Galactose Oxidase Variant

Mattey, Ashley P.; Birmingham, William R.; Both, Peter; Kreß, Nico; Huang, Kun; Munster, Jolanda M. van; Gs, Bulmer; Parmeggiani, Fabio; Voglmeir, Josef; Martínez, Juana Elizabeth Reyes; Turner, Nicholas J.; Flitsch, Sabine L.

doi:10.1021/acscatal.9b02873

Cited by 17 publications

(13 citation statements)

References 32 publications

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“…Galactose Although the molecular basis for the specificity difference between Fgr AAO and Fox AAO and the archetypal Fgr GalOx is unclear in the absence of any three-dimensional substrate or product complexes of AA5 members [ 17 – 19 , 41 , 42 , 47 , 57 , 79 , 80 ], sequence analysis and molecular modeling provides some insight. In particular, there is literature precedent for docking ligand structures into models of AA5_2 variants for activity analysis [ 40 , 81 ].…”

Section: Resultsmentioning

confidence: 99%

Two Fusarium copper radical oxidases with high activity on aryl alcohols

Cleveland

Lafond

Xia

et al. 2021

Biotechnol Biofuels

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Background Biomass valorization has been suggested as a sustainable alternative to petroleum-based energy and commodities. In this context, the copper radical oxidases (CROs) from Auxiliary Activity Family 5/Subfamily 2 (AA5_2) are attractive biocatalysts for the selective oxidation of primary alcohols to aldehydes. Originally defined by the archetypal galactose 6-oxidase from Fusarium graminearum, fungal AA5_2 members have recently been shown to comprise a wide range of specificities for aromatic, aliphatic and furan-based alcohols. This suggests a broader substrate scope of native CROs for applications. However, only 10% of the annotated AA5_2 members have been characterized to date. Results Here, we define two homologues from the filamentous fungi Fusarium graminearum and F. oxysporum as predominant aryl alcohol oxidases (AAOs) through recombinant production in Pichia pastoris, detailed kinetic characterization, and enzyme product analysis. Despite possessing generally similar active-site architectures to the archetypal FgrGalOx, FgrAAO and FoxAAO have weak activity on carbohydrates, but instead efficiently oxidize specific aryl alcohols. Notably, both FgrAAO and FoxAAO oxidize hydroxymethyl furfural (HMF) directly to 5-formyl-2-furoic acid (FFCA), and desymmetrize the bioproduct glycerol to the uncommon L-isomer of glyceraldehyde. Conclusions This work expands understanding of the catalytic diversity of CRO from AA5_2 to include unique representatives from Fusarium species that depart from the well-known galactose 6-oxidase activity of this family. Detailed enzymological analysis highlights the potential biotechnological applications of these orthologs in the production of renewable plastic polymer precursors and other chemicals.

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Section: Resultsmentioning

confidence: 99%

Two Fusarium copper radical oxidases with high activity on aryl alcohols

Cleveland

Lafond

Xia

et al. 2021

Biotechnol Biofuels

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“…114 It could also be immobilized on Purolite for the oxidation of benzyl alcohols and recycled, 115,116 exploited in the chemoenzymatic synthesis of 3-deoxy-3-fluorofucose, 117 or used in the generation of DNA-encoded libraries. 118 Other developments involve glycoprotein labeling, 114 oxidation of Neu5Gc-containing glycans, 119 or even the labeling of polysaccharides. 120 Christophe Biot (University of Lille) then reported on the use of the Chemical Reporter Strategy to image sialylation in live cells.…”

Section: ■ Daymentioning

confidence: 99%

ICBS 2021: Looking Toward the Next Decade of Chemical Biology

2022

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“…As for PAL, oxidation is followed by an oxime ligation step that is accelerated in the presence of an aniline catalyst [ 24 ]. Specificity for other types of monosaccharides has been achieved by the generation of engineered variants of galactose oxidase that target, for example, N -glycolylneuraminic acid (Neu5Gc) [ 25 ].…”

Section: Oxidation Of Cell Surface Glycansmentioning

confidence: 99%

Chemical reporters to study mammalian O-glycosylation

Huxley

Willems

2021

Biochemical Society Transactions

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Glycans play essential roles in a range of cellular processes and have been shown to contribute to various pathologies. The diversity and dynamic nature of glycan structures and the complexities of glycan biosynthetic pathways make it challenging to study the roles of specific glycans in normal cellular function and disease. Chemical reporters have emerged as powerful tools to characterise glycan structures and monitor dynamic changes in glycan levels in a native context. A variety of tags can be introduced onto specific monosaccharides via the chemical modification of endogenous glycan structures or by metabolic or enzymatic incorporation of unnatural monosaccharides into cellular glycans. These chemical reporter strategies offer unique opportunities to study and manipulate glycan functions in living cells or whole organisms. In this review, we discuss recent advances in metabolic oligosaccharide engineering and chemoenzymatic glycan labelling, focusing on their application to the study of mammalian O-linked glycans. We describe current barriers to achieving glycan labelling specificity and highlight innovations that have started to pave the way to overcome these challenges.

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Selective Oxidation of N-Glycolylneuraminic Acid Using an Engineered Galactose Oxidase Variant

Cited by 17 publications

References 32 publications

Two Fusarium copper radical oxidases with high activity on aryl alcohols

Two Fusarium copper radical oxidases with high activity on aryl alcohols

ICBS 2021: Looking Toward the Next Decade of Chemical Biology

Chemical reporters to study mammalian O-glycosylation

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