Engineering the specificity of trehalose phosphorylase as a general strategy for the production of glycosyl phosphates

Chen, Chao; Borght, Jef Van der; Vreese, Rob De; D’hooghe, Matthias; Soetaert, Wim; Desmet, Tom

doi:10.1039/c4cc02202e

Cited by 11 publications

(7 citation statements)

References 36 publications

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“…Interestingly, the Desmet group is also developing enzymatic approaches to synthesize glycosyl phosphates such as β-D-glucose-1-phosphate, which, if efficient, could alleviate this problem. 72,73 On the other hand, retaining TreP enzymes, which can produce trehalose from glucose and much cheaper α-D-glucose-1-phosphate ($25/g), may provide a more efficient route to trehalose analogues. Unfortunately, to date all retaining TrePs that have been evaluated have shown poor stability and expression yields, and furthermore exhibited low substrate promiscuity.…”

Section: Approaches To Trehalose Analogue Synthesismentioning

confidence: 99%

Tailoring trehalose for biomedical and biotechnological applications

O'Neill

Piligian

Olson

et al. 2017

Pure and Applied Chemistry

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Trehalose is a non-reducing sugar whose ability to stabilize biomolecules has brought about its widespread use in biological preservation applications. Trehalose is also an essential metabolite in a number of pathogens, most significantly the global pathogen Mycobacterium tuberculosis, though it is absent in humans and other mammals. Recently, there has been a surge of interest in modifying the structure of trehalose to generate analogues that have applications in biomedical research and biotechnology. Non-degradable trehalose analogues could have a number of advantages as bioprotectants and food additives. Trehalose-based imaging probes and inhibitors are already useful as research tools and may have future value in the diagnosis and treatment of tuberculosis, among other uses. Underlying the advancements made in these areas are novel synthetic methods that facilitate access to and evaluation of trehalose analogues. In this review, we focus on both aspects of the development of this class of molecules. First, we consider the chemical and chemoenzymatic methods that have been used to prepare trehalose analogues and discuss their prospects for synthesis on commercially relevant scales. Second, we describe ongoing efforts to develop and deploy detectable trehalose analogues, trehalose-based inhibitors, and non-digestible trehalose analogues. The current and potential future uses of these compounds are discussed, with an emphasis on their roles in understanding and combatting mycobacterial infection.

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Section: Approaches To Trehalose Analogue Synthesismentioning

confidence: 99%

Tailoring trehalose for biomedical and biotechnological applications

O'Neill

Piligian

Olson

et al. 2017

Pure and Applied Chemistry

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“…In the C. subterraneus TP active site, the three mutagenized positions (L694G/A693Q/W371Y) causing a switch in donor specificity are indicated in green with the corresponding wild-type residues shown in yellow. The lactotrehalose substrate is indicated in orange [ 66 , 67 ].…”

Section: Biocatalytic Synthesis Routes For Trehalose Analoguesmentioning

confidence: 99%

“…In addition to focusing on the acceptor flexibility, engineering the donor specificity of an inverted TP could further extend the pool of trehalose analogues that could be produced with these enzymes. To give an idea of the potential, the donor specificity of the C. subterraneus TP could be modified towards β-Gal-1P by introducing three mutations (L694G/A693Q/W371Y), identified through iterative saturation ( Figure 6 ) [ 67 ]. The triple mutant displayed a 10-fold improved catalytic efficiency on the alternative donor substrate, and could thus be used for the synthesis of lactotrehalose, with d -glucose as acceptor, and even galactotrehalose, with d -galactose as acceptor.…”

Section: Biocatalytic Synthesis Routes For Trehalose Analoguesmentioning

confidence: 99%

“…The triple mutant displayed a 10-fold improved catalytic efficiency on the alternative donor substrate, and could thus be used for the synthesis of lactotrehalose, with d -glucose as acceptor, and even galactotrehalose, with d -galactose as acceptor. Interestingly, the combined action of the T. brockii and the C. subterraneus variants can also be used for the net phosphorylation of d -galactose, which then enters such a two-step process from the acceptor side, but leaves from the donor side with lactotrehalose as intermediate ( Figure 6 ) [ 67 ].…”

Section: Biocatalytic Synthesis Routes For Trehalose Analoguesmentioning

confidence: 99%

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Trehalose Analogues: Latest Insights in Properties and Biocatalytic Production

Walmagh

Zhao

Desmet

2015

IJMS

Self Cite

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Trehalose (α-d-glucopyranosyl α-d-glucopyranoside) is a non-reducing sugar with unique stabilizing properties due to its symmetrical, low energy structure consisting of two 1,1-anomerically bound glucose moieties. Many applications of this beneficial sugar have been reported in the novel food (nutricals), medical, pharmaceutical and cosmetic industries. Trehalose analogues, like lactotrehalose (α-d-glucopyranosyl α-d-galactopyranoside) or galactotrehalose (α-d-galactopyranosyl α-d-galactopyranoside), offer similar benefits as trehalose, but show additional features such as prebiotic or low-calorie sweetener due to their resistance against hydrolysis during digestion. Unfortunately, large-scale chemical production processes for trehalose analogues are not readily available at the moment due to the lack of efficient synthesis methods. Most of the procedures reported in literature suffer from low yields, elevated costs and are far from environmentally friendly. “Greener” alternatives found in the biocatalysis field, including galactosidases, trehalose phosphorylases and TreT-type trehalose synthases are suggested as primary candidates for trehalose analogue production instead. Significant progress has been made in the last decade to turn these into highly efficient biocatalysts and to broaden the variety of useful donor and acceptor sugars. In this review, we aim to provide an overview of the latest insights and future perspectives in trehalose analogue chemistry, applications and production pathways with emphasis on biocatalysis.

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“…PG's glycosidic bond is easily hydrolysed at high acid levels, and tetrapeptides can be weakly denaturated under high temperatures. In recent years, phosphate, which is inexpensive, can be conveniently obtained and can be used under mild reaction conditions and has been developed into a new generation of optimal reagents to perform phosphorylation (Chen et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Phosphorylation of peptidoglycan from Lactobacillus acidophilus and its immunoregulatory function

Pan

Sun

et al. 2016

Int J of Food Sci Tech

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Summary Peptidoglycan (PG) is available from a wide variety of lactic acid bacteria (LAB) and is the main structure of cell wall components. Phosphorylated modification would bring new properties such as the potential antioxidant activities and antiviral capability to an organic molecule. In the present work, small molecular fragments of PG (derived from Lactobacillus acidophilus) hydrolysed by mutanolysin were phosphorylated under optimal conditions. P‐PG had a monomer molecular structure of GlcNAC[PO3]–MurNAC–Ala–Glu–Lys–Ala, with a molecular mass of 884 Da and a phosphorus content of 8.9%. P‐PG displayed some immunoregulatory activity in lipopolysaccharides (LPS) stimulated RAW 264.7 macrophages. Compared with the LPS‐stimulated group, the addition of P‐PG inhibited the secretion of GM‐CSF, TNF‐α and IL‐1 in a dose‐dependent manner. The effect of 50 μg mL−1 of P‐PG was more significant than 50 μg mL−1 of PG. Lower fluorescence of lysosomes was observed in P‐PG‐treated RAW 264.7 cells may also reveal some immune defence function in the LPS‐induced macrophages.

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Engineering the specificity of trehalose phosphorylase as a general strategy for the production of glycosyl phosphates

Cited by 11 publications

References 36 publications

Tailoring trehalose for biomedical and biotechnological applications

Tailoring trehalose for biomedical and biotechnological applications

Trehalose Analogues: Latest Insights in Properties and Biocatalytic Production

Phosphorylation of peptidoglycan from Lactobacillus acidophilus and its immunoregulatory function

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