Enzymatic production of α-d-galactose 1-phosphate by lactose phosphorolysis

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

doi:10.1007/s10529-009-0087-1

Cited by 22 publications

(7 citation statements)

References 20 publications

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“…Using permeabilized E. coli cells, the engineered "lactose phosphorylase" was employed in the synthesis of expensive α-D-galactose 1-phosphate from low-cost substrates such as lactose and phosphate, lactose + phosphate → α-D-galactose 1-phosphate + glucose. Highly pure α-D-galactose 1-phosphate (9.5 g) was obtained from a 1-L reaction mixture (500 mM lactose, 50 mM phosphate at 50°C for 48 h) after anion exchange chromatography and ethanol precipitation [92].…”

Section: Cellobiose Phosphorylasementioning

confidence: 99%

Carbohydrate synthesis by disaccharide phosphorylases: Reactions, catalytic mechanisms and application in the glycosciences

Luley‐Goedl

Nidetzky

2010

Biotechnology Journal

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Disaccharide phosphorylases are glycosyltransferases (EC 2.4.1.α) of specialized carbohydrate metabolism in microorganisms. They catalyze glycosyl transfer to phosphate using a disaccharide as donor substrate. Phosphorylases for the conversion of naturally abundant disaccharides including sucrose, maltose, α,α‐trehalose, cellobiose, chitobiose, and laminaribiose have been described. Structurally, these disaccharide phosphorylases are often closely related to glycoside hydrolases and transglycosidases. Mechanistically, they are categorized according the stereochemical course of the reaction catalyzed, whereby the anomeric configuration of the disaccharide donor substrate may be retained or inverted in the sugar 1‐phosphate product. Glycosyl transfer with inversion is thought to occur through a single displacement‐like catalytic mechanism, exemplified by the reaction coordinate of cellobiose/chitobiose phosphorylase. Reaction via configurational retention takes place through the double displacement‐like mechanism employed by sucrose phosphorylase. Retaining α,α‐trehalose phosphorylase (from fungi) utilizes a different catalytic strategy, perhaps best described by a direct displacement mechanism, to achieve stereochemical control in an overall retentive transformation. Disaccharide phosphorylases have recently attracted renewed interest as catalysts for synthesis of glycosides to be applied as food additives and cosmetic ingredients. Relevant examples are lacto‐N‐biose and glucosylglycerol whose enzymatic production was achieved on multikilogram scale. Protein engineering of phosphorylases is currently pursued in different laboratories with the aim of broadening the donor and acceptor substrate specificities of naturally existing enzyme forms, to eventually generate a toolbox of new catalysts for glycoside synthesis.

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Section: Cellobiose Phosphorylasementioning

confidence: 99%

Carbohydrate synthesis by disaccharide phosphorylases: Reactions, catalytic mechanisms and application in the glycosciences

Luley‐Goedl

Nidetzky

2010

Biotechnology Journal

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“…It has been described in literature that E. coli phosphatase activity could be lowered significantly by using an agp-negative strain like CGSC8974 and growing it in LB medium, supplemented with glucose and phosphate [13,27,28]. This effect was quantified and the specificity of E. coli phosphatases for αGlc1P and βGlc1P was evaluated (Fig.…”

Section: Production Of the Biocatalystsmentioning

confidence: 99%

“…Glycoside phosphorylases, in contrast, only need inorganic phosphate to produce a glycosylphosphate. In literature, the synthesis of α-glucose-1-phosphate (αGlc1P) from starch [10,11] or sucrose [12] and that of α-galactose-1-phosphate (αGal1P) from lactose [13,14] have previously been reported. A production process for βGlc1P has not yet been described in detail, but is similarly feasible using GH65 disaccharide phosphorylases from cheap substrates [15] briefly mention the production of βGlc1P from trehalose, using typical reaction conditions employed for αGlc1P [16].…”

Section: Introductionmentioning

confidence: 99%

Enzymatic production of β‐D‐glucose‐1‐phosphate from trehalose

Borght¹,

Desmet²,

Soetaert³

2010

Biotechnology Journal

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beta-D-Glucose-1-phosphate (beta Glc1P) is an efficient glucosyl donor for both enzymatic and chemical glycosylation reactions but is currently very costly and not available in large amounts. This article provides an efficient production method of beta Glc1P from trehalose and phosphate using the thermostable trehalose phosphorylase from Thermoanaerobacter brockii. At the process temperature of 60 C, Escherichia coli expression host cells are lysed and cell treatment prior to the reaction is, therefore, not required. In this way, the theoretical maximum yield of 26% could be easily achieved. Two different purification strategies have been compared, anion exchange chromatography or carbohydrate removal by treatment with trehalase and yeast, followed by chemical phosphate precipitation. In a next step, beta Glc1P was precipitated with ethanol but this did not induce crystallization, in contrast to what ist observed with other glycosylphosphates. After conversion of the product to its cyclohexylammonium salt, however, crystals could be readily obtained. Although both purification methods were quantitative (>99% recovery), a large amount of product (50%) was lost during crystallization. Nevertheless, a production process for crystalline beta Glc1P is now available from the cheap substrates trehalose and inorganic phosphate

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“…1 H NMR signals were assigned with the aid of gDQCOSY. 13 C NMR signals were assigned with the aid of gHSQCAD. Coupling constants are reported in Hertz.…”

Section: Generalmentioning

confidence: 99%

“…They serve as precursors to sugar-nucleotides [6,7], the sugar-donor components utilised by glycosyltransferases in the assembly of oligosaccharides and glycans and have played a key role in the development of glycosylated natural-product-based therapeutics [8]. Additionally, glycosyl 1-phosphates have been used as substrates for glycoside phosphorylases (a rapidly expanding family of CAZy enzymes) for the synthesis of oligosaccharide targets [9,10] and also play important technological roles in the food and detergent sectors [11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

6R/S-deutero-α-d-mannopyranoside 1-phosphate

Ahmadipour

Miller

2019

Molbank

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6R/S-deutero-α-d-mannopyranoside 1-phosphate was synthesised from a C6 aldehydic mannose thioglycoside donor in four steps. Using NaBD4 as the reductant, isotopic enrichment at C6 was achieved and the resultant C6-deuterated material was converted through to the glycosyl 1-phosphate using a protection/glycosylation/deprotection sequence. The product was fully characterised by 1H, 13C, 31P and 2D NMR, alongside MS analysis.

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Enzymatic production of α-d-galactose 1-phosphate by lactose phosphorolysis

Cited by 22 publications

References 20 publications

Carbohydrate synthesis by disaccharide phosphorylases: Reactions, catalytic mechanisms and application in the glycosciences

Carbohydrate synthesis by disaccharide phosphorylases: Reactions, catalytic mechanisms and application in the glycosciences

Enzymatic production of β‐D‐glucose‐1‐phosphate from trehalose

6R/S-deutero-α-d-mannopyranoside 1-phosphate

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