Glycoside phosphorylases: Structure, catalytic properties and biotechnological potential

Puchart, Vladimı́r

doi:10.1016/j.biotechadv.2015.02.002

Cited by 85 publications

(76 citation statements)

References 157 publications

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“…Recent review articles (O'Neill and Field 2015;Puchart 2015) have described most of the uses of phosphorylases. Here, we describe the recent reported studies on the use of phosphorylases for preparing sugars.…”

Section: Preparation Of Glycosides By Phosphorylases (Recent Publicatmentioning

confidence: 99%

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Diversity of phosphorylases in glycoside hydrolase families

Kitaoka

2015

Appl Microbiol Biotechnol

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Phosphorylases are useful catalysts for the practical preparation of various sugars. The number of known specificities was 13 in 2002 and is now 30. The drastic increase in available genome sequences has facilitated the discovery of novel activities. Most of these novel phosphorylase activities have been identified through the investigations of glycoside hydrolase families containing known phosphorylases. Here, the diversity of phosphorylases in each family is described in detail.

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Section: Preparation Of Glycosides By Phosphorylases (Recent Publicatmentioning

confidence: 99%

“…O' Neill and Field (2015) tabulated the uses of phosphorylases for the syntheses of glycosides. Most recently, Puchart (2015) provided a good overview on phosphorylase research. In the present minireview, we focus on the diversity of phosphorylase activities.…”

Section: Introductionmentioning

confidence: 99%

Diversity of phosphorylases in glycoside hydrolase families

Kitaoka

2015

Appl Microbiol Biotechnol

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“…A three‐enzyme cascade reaction converts sucrose and glucose into soluble cellodextrins. The enzymes used are glucoside phosphorylases . A common feature of their reactions is the reversible glucosyl transfer to phosphate, giving α‐ d ‐glucose 1‐phosphate (αGlc1‐ P ) as the product.…”

Section: Introductionmentioning

confidence: 99%

Three‐Enzyme Phosphorylase Cascade for Integrated Production of Short‐Chain Cellodextrins

Zhong

Nidetzky

2019

Biotechnology Journal

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Cellodextrins are linear β‐1,4‐gluco‐oligosaccharides that are soluble in water up to a degree of polymerization (DP) of ≈6. Soluble cellodextrins have promising applications as nutritional ingredients. A DP‐controlled, bottom‐up synthesis from expedient substrates is desired for their bulk production. Here, a three‐enzyme glycoside phosphorylase cascade is developed for the conversion of sucrose and glucose into short‐chain (soluble) cellodextrins (DP range 3–6). The cascade reaction involves iterative β‐1,4‐glucosylation of glucose from α‐glucose 1‐phosphate (αGlc1‐P) donor that is formed in situ from sucrose and phosphate. With final concentration and yield of the soluble cellodextrins set as targets for biocatalytic synthesis, three major factors of reaction efficiency are identified and partly optimized: the ratio of enzyme activity, the ratio of sucrose and glucose, and the phosphate concentration used. The efficient use of the phosphate/αGlc1‐P shuttle for cellodextrin production is demonstrated and the soluble product at 40 g L−1 is obtained under near‐complete utilization of the donor substrate offered (88 mol% from 200 mm sucrose). The productivity is 16 g (L h)−1. Through a simple two‐step route, the soluble cellodextrins are recovered from the reaction mixture in ≥95% purity and ≈92% yield. Overall, this study provides the basis for their integrated production.

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“…Glycoside phosphorylases (GPs) are a group of carbohydrate‐active enzymes that catalyse the reversible cleavage of glycosidic linkages in di‐ or oligosaccharides by transferring the non‐reducing end glycosyl residue to inorganic phosphate . The reverse reaction (synthetic reaction) of GPs is of practical importance because it can be used as an alternative method of enzymatic glycosylation by utilising sugar 1‐phosphates as donor substrates.…”

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

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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.

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Glycoside phosphorylases: Structure, catalytic properties and biotechnological potential

Cited by 85 publications

References 157 publications

Diversity of phosphorylases in glycoside hydrolase families

Diversity of phosphorylases in glycoside hydrolase families

Three‐Enzyme Phosphorylase Cascade for Integrated Production of Short‐Chain Cellodextrins

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

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