Engineering of cellobiose phosphorylase for glycoside synthesis

Groeve, Manu R.M. De; Desmet, Tom; Soetaert, Wim

doi:10.1016/j.jbiotec.2011.07.006

Cited by 16 publications

(12 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…De Groeve et al [147] engineered both the donor and acceptor specificities of a cellobiose phosphorylase (CP), first discovered in extracts from the bacterium Clostridium thermocellum and belonging to the GH94 family. This enzyme catalyses the reversible phosphorolysis of cellobiose into a-D-glucosyl 1-phosphate (a-D-Glc-1P).…”

Section: Production Of Oligosaccharidesmentioning

confidence: 99%

See 1 more Smart Citation

Harnessing glycoenzyme engineering for synthesis of bioactive oligosaccharides

et al. 2019

View full text Add to dashboard Cite

Combined with chemical synthesis, the use of glycoenzyme biocatalysts has shown great synthetic potential over recent decades owing to their remarkable versatility in terms of substrates and regio- and stereoselectivity that allow structurally controlled synthesis of carbohydrates and glycoconjugates. Nonetheless, the lack of appropriate enzymatic tools with requisite properties in the natural diversity has hampered extensive exploration of enzyme-based synthetic routes to access relevant bioactive oligosaccharides, such as cell-surface glycans or prebiotics. With the remarkable progress in enzyme engineering, it has become possible to improve catalytic efficiency and physico-chemical properties of enzymes but also considerably extend the repertoire of accessible catalytic reactions and tailor novel substrate specificities. In this review, we intend to give a brief overview of the advantageous use of engineered glycoenzymes, sometimes in combination with chemical steps, for the synthesis of natural bioactive oligosaccharides or their precursors. The focus will be on examples resulting from the three main classes of glycoenzymes specialized in carbohydrate synthesis: glycosyltransferases, glycoside hydrolases and glycoside phosphorylases.

show abstract

Section: Production Of Oligosaccharidesmentioning

confidence: 99%

“…. which is particularly valuable for glycoside synthesis [147]. To further expand the acceptor specificity, De Groeve et al [149] identified a residue (E649) in the structure of CP from C. uda, whose side chain has hydrogen bonding with the D-Glc acceptor.…”

Section: Production Of Oligosaccharidesmentioning

confidence: 99%

Harnessing glycoenzyme engineering for synthesis of bioactive oligosaccharides

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Structure‐guided site‐directed mutagenesis has been performed extensively on CBP from Cellvibrio gilvus ( Cg CBP), including its conversion into a lactose phosphorylase . In addition, a single mutation (E649C) in Cg CBP created an enzyme variant capable of using methyl β‐glucoside, ethyl β‐glucoside, and phenyl β‐glucoside as acceptors . Another Cg CBP variant was created by mutation of five amino acids within and around the entrance to the enzyme active site, which broadened the acceptor range to include both β‐ and α‐glucosides .…”

Section: Introductionmentioning

confidence: 99%

Unravelling the Specificity of Laminaribiose Phosphorylase from Paenibacillus sp. YM‐1 towards Donor Substrates Glucose/Mannose 1‐Phosphate by Using X‐ray Crystallography and Saturation Transfer Difference NMR Spectroscopy

et al. 2018

View full text Add to dashboard Cite

Glycoside phosphorylases (GPs) carry out a reversible phosphorolysis of carbohydrates into oligosaccharide acceptors and the corresponding sugar 1-phosphates. The reversibility of the reaction enables the use of GPs as biocatalysts for carbohydrate synthesis. Glycosyl hydrolase family 94 (GH94), which only comprises GPs, is one of the most studied GP families that have been used as biocatalysts for carbohydrate synthesis, in academic research and in industrial production. Understanding the mechanism of GH94 enzymes is a crucial step towards enzyme engineering to improve and expand the applications of these enzymes in synthesis. In this work with a GH94 laminaribiose phosphorylase from Paenibacillus sp. YM-1 (PsLBP), we have demonstrated an enzymatic synthesis of disaccharide 1 (β-d-mannopyranosyl-(1→3)-d-glucopyranose) by using a natural acceptor glucose and noncognate donor substrate α-mannose 1-phosphate (Man1P). To investigate how the enzyme recognises different sugar 1-phosphates, the X-ray crystal structures of PsLBP in complex with Glc1P and Man1P have been solved, providing the first molecular detail of the recognition of a noncognate donor substrate by GPs, which revealed the importance of hydrogen bonding between the active site residues and hydroxy groups at C2, C4, and C6 of sugar 1-phosphates. Furthermore, we used saturation transfer difference NMR spectroscopy to support crystallographic studies on the sugar 1-phosphates, as well as to provide further insights into the PsLBP recognition of the acceptors and disaccharide products.

show abstract

“…Relatively narrow substrate specificity prevents wide-range applications of disaccharide phosphorylases for glycoside synthesis (De Groeve et al 2010a ). Nonetheless, cellobiose and laminaribiose phosphorylase can be used for β-glucosylation or β-galactosylation of various molecules (Table 2 ) (De Groeve et al 2010a ; De Winter et al 2015b ). Lower affinity, acceptor and donor specificity, or specific activity of these enzymes related to non-natural substrates could be improved via enzyme engineering.…”

Section: Engineering Of β-Glucobiose and β-Glucan Phosphorylasesmentioning

confidence: 99%

β-Glucan phosphorylases in carbohydrate synthesis

Ubiparip

Doncker

Beerens

et al. 2021

Appl Microbiol Biotechnol

Self Cite

View full text Add to dashboard Cite

β-Glucan phosphorylases are carbohydrate-active enzymes that catalyze the reversible degradation of β-linked glucose polymers, with outstanding potential for the biocatalytic bottom-up synthesis of β-glucans as major bioactive compounds. Their preference for sugar phosphates (rather than nucleotide sugars) as donor substrates further underlines their significance for the carbohydrate industry. Presently, they are classified in the glycoside hydrolase families 94, 149, and 161 (www.cazy.org). Since the discovery of β-1,3-oligoglucan phosphorylase in 1963, several other specificities have been reported that differ in linkage type and/or degree of polymerization. Here, we present an overview of the progress that has been made in our understanding of β-glucan and associated β-glucobiose phosphorylases, with a special focus on their application in the synthesis of carbohydrates and related molecules. Key points • Discovery, characteristics, and applications of β-glucan phosphorylases. • β-Glucan phosphorylases in the production of functional carbohydrates.

show abstract

Engineering of cellobiose phosphorylase for glycoside synthesis

Cited by 16 publications

References 42 publications

Harnessing glycoenzyme engineering for synthesis of bioactive oligosaccharides

Harnessing glycoenzyme engineering for synthesis of bioactive oligosaccharides

Unravelling the Specificity of Laminaribiose Phosphorylase from Paenibacillus sp. YM‐1 towards Donor Substrates Glucose/Mannose 1‐Phosphate by Using X‐ray Crystallography and Saturation Transfer Difference NMR Spectroscopy

β-Glucan phosphorylases in carbohydrate synthesis

Contact Info

Product

Resources

About