Towards the synthesis of glycosylated dihydrochalcone natural products using glycosyltransferase-catalysed cascade reactions

Gutmann, Alexander; Bungaruang, Linda; Weber, Hedda K.; Leypold, Mario; Breinbauer, Rolf; Nidetzky, Bernd

doi:10.1039/c4gc00960f

Cited by 52 publications

(44 citation statements)

References 60 publications

(116 reference statements)

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“…Blue box: seemingly futile cycle of Suc metabolism. Gene abbreviations: sps=sucrose-phosphate synthase, spp=sucrose-phosphate phosphatase, susy=sucrose synthase, sp=sucrose phosphorylase, frk=fructokinase, pfkb=PfkB family of carbohydrate kinase, amsA= amylosucrase abundant and cheap substrate sucrose and for the costeffective glycosylation of small molecules by coupling to a GT (Zervosen et al 1998;Masada et al 2007;Son et al 2009;Terasaka et al 2012;Bungaruang et al 2013;Gutmann et al 2014). Key requirements for successful application in industry predominantly consist of highly active, (thermo) stable enzymes, and a high number of regeneration cycles for UDP (De Bruyn et al 2015).…”

Section: Industrial Applicationsmentioning

confidence: 98%

“…In 1993, Elling and coworkers demonstrated the production of expensive nucleotide sugars (NDP-Glc) starting from the abundant and cheap substrate Suc. Moreover, SuSy can also be coupled with a glycosyltransferase (GT), which has resulted in a cost-effective method for the glycosylation of small molecules (Brinkmann et al 2001;Masada et al 2007;Son et al 2009;Terasaka et al 2012;Bungaruang et al 2013;Gutmann et al 2014). However, low activities and poor stability of the reported SuSy enzymes have impeded their commercial exploitation so far.…”

Section: Introductionmentioning

confidence: 97%

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Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Diricks

Bruyn

Daele

et al. 2015

Appl Microbiol Biotechnol

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Sucrose synthase (SuSy) catalyzes the reversible conversion of sucrose and a nucleoside diphosphate into fructose and nucleotide (NDP)-glucose. To date, only SuSy's from plants and cyanobacteria, both photosynthetic organisms, have been characterized. Here, four prokaryotic SuSy enzymes from the nonphotosynthetic organisms Nitrosomonas Europaea (SuSyNe), Acidithiobacillus caldus (SuSyAc), Denitrovibrio acetiphilus (SusyDa), and Melioribacter roseus (SuSyMr) were recombinantly expressed in Escherichia coli and thoroughly characterized. The purified enzymes were found to display high-temperature optima (up to 80 °C), high activities (up to 125 U/mg), and high thermostability (up to 15 min at 60 °C). Furthermore, SuSyAc, SuSyNe, and SuSyDa showed a clear preference for ADP as nucleotide, as opposed to plant SuSy's which prefer UDP. A structural and mutational analysis was performed to elucidate the difference in NDP preference between eukaryotic and prokaryotic SuSy's. Finally, the physiological relevance of this enzyme specificity is discussed in the context of metabolic pathways and genomic organization.

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Section: Industrial Applicationsmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 97%

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Diricks

Bruyn

Daele

et al. 2015

Appl Microbiol Biotechnol

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“…The obtained maltosyl-neohesperidin dihydrochalcone was 700 times more soluble in water but was, however, less sweet than the substrate, whereas Eichenberger et al [62] proposed to employ S. cerevisiae as a microbial cell factory for de novo production of various dihydrochalcones of commercial interest. In turn, Gutmann et al [63] described an efficient synthesis of glycosylated dihydrochalcones like phlorizin (2), davidigenin, and confusoside, using glycosyltransferase-catalysed cascade reactions.…”

Section: Chemical Synthesis Of Dihydrochalconesmentioning

confidence: 99%

Dihydrochalcones: Methods of Acquisition and Pharmacological Properties—A First Systematic Review

2019

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Dihydrochalcones are a class of secondary metabolites, for which demand in biological and pharmacological applications is still growing. They posses several health-endorsing properties and, therefore, are promising candidates for further research and development. However, low content of dihydrochalcones in plants along with their low solubility and bioavailability restrict the development of these compounds as clinical therapeutics. Therefore, chemomicrobial and enzymatic modifications are required to expand their application. This review aims at analyzing and summarizing the methods of obtaining dihydrochalcones and of presenting their pharmacological actions that have been described in the literature to support potential future development of this group of compounds as novel therapeutic drugs. We have also performed an evaluation of the available literature on beneficial effects of dihydrochalcones with potent antioxidant activity and multifactorial pharmacological effects, including antidiabetic, antitumor, lipometabolism regulating, antioxidant, anti-inflammatory, antibacterial, antiviral, and immunomodulatory ones. In addition, we provide useful information on their properties, sources, and usefulness in medicinal chemistry.

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“…The specificity of SuSy for NDP is relatively broad, thus enabling synthesis of the corresponding NDP-glucoses [12,14]. However, because the majority of the known glucosyltransferases requires UDP-glucose, this was also the clear focus of past efforts of using SuSy for donor substrate recycling [15][16][17][18][19][20]. To expand the scope of SuSy-GT cascade reactions beyond glucosyl transfer, additional NDP-glucose-modifying enzymes were added to enable regeneration of UDP-galactose [21][22][23][24][25], UDP-glucuronic acid [26] and dTDP-rhamnose [27].…”

Section: Introductionmentioning

confidence: 99%

Glycosyltransferase cascades for natural product glycosylation: Use of plant instead of bacterial sucrose synthases improves the UDP‐glucose recycling from sucrose and UDP

Gutmann

Lepak

Diricks

et al. 2017

Biotechnology Journal

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Natural product glycosylations by Leloir glycosyltransferases (GTs) require expensive nucleotide-activated sugars as substrates. Sucrose synthase (SuSy) converts sucrose and uridine 5'-diphosphate (UDP) into UDP-glucose. Coupling of SuSy and GT reactions in one-pot cascade transformations creates a UDP cycle, which regenerates the UDP-glucose continuously and so makes it an expedient donor for glucoside production. Here we compare SuSys with divergent kinetic characteristics for UDP-glucose recycling in the synthesis of the natural C-glucoside nothofagin. Development of a fast reversed-phase ion-pairing HPLC method, quantifying all relevant reactants from the coupled conversion in a single run, was key to dissect the main factors of recycling efficiency. Limitations due to high K , both for UDP and sucrose, were revealed for the bacterial SuSy from Acidithiobacillus caldus. The L637M-T640V double mutant of this SuSy with a 60-fold reduced KM for UDP substantially improved UDP-glucose recycling. The SuSy from Glycine max (soybean) was nevertheless the most active enzyme at the UDP (≤ 0.5 mM) and sucrose (≤ 1 M) concentrations used. It was also unexpectedly stable at up to 50°C where spontaneous decomposition of UDP-glucose started to become problematic. The herein gained in-depth understanding of requirements for UDP-glucose regeneration supports development of efficient GT-SuSy cascades.

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Towards the synthesis of glycosylated dihydrochalcone natural products using glycosyltransferase-catalysed cascade reactions

Abstract: Regioselective 2′-O- and 4′-O-β-d-glucosylation of dihydrochalcones was achieved through glycosyltransferase cascade reactions using glucosyl donor substrate supply from sucrose.

Cited by 52 publications

References 60 publications

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Dihydrochalcones: Methods of Acquisition and Pharmacological Properties—A First Systematic Review

Glycosyltransferase cascades for natural product glycosylation: Use of plant instead of bacterial sucrose synthases improves the UDP‐glucose recycling from sucrose and UDP

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