An Enzyme Module System for the Synthesis of dTDP‐activated Deoxysugars from dTMP and Sucrose

Elling, Lothar; Rupprath, Carsten; Günther, Nicole; Römer, Ulrike; Verseck, Stefan; Weingarten, P.; Dräger, Gerald; Kirschning, Andreas; Piepersberg, Wolfgang

doi:10.1002/cbic.200500037

Cited by 41 publications

(37 citation statements)

References 72 publications

(70 reference statements)

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“…The synthesis of 34 from sucrose and dTMP (108) was realized by the combination of the SuSy module and the deoxysugar module containing the RmlB-D enzymes (Fig. (15), [230]. A two-step strategy avoiding feed back inhibition by the product resulted in 19.1 mg of 34 with an overall yield of 46.6% (Tab.…”

Section: Chemo-enzymatic Synthesismentioning

confidence: 99%

“…A preparative synthesis of 14 was carried out by Oh et al [99]. The synthesis was started from dTMP (108) by the The described enzyme system has recently been further developed towards a flexible enzyme module system facilitating the synthesis of dTDP-deoxyhexoses [230]. The combination of recombinant enzymes in the SuSy and the deoxysugar module was utilized for the synthesis of 14 from dTMP (0.31 mmol, 183.5 mg, 72.2% overall yield) (Fig.…”

Section: Chemo-enzymatic Synthesismentioning

confidence: 99%

“…Starting from 41 and dTMP (108), 13 was synthesized in an overall yield of 62% (with reference to dTMP) which corresponds to 430 µmol (253 mg) [230].…”

Section: Chemo-enzymatic Synthesismentioning

confidence: 99%

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Nucleotide Deoxysugars: Essential Tools for the Glycosylation Engineering of Novel Bioactive Compounds

Rupprath¹,

Schumacher²,

Elling

2005

CMC

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The irreversible spread of new resistance mechanisms against existing therapeutical antibiotics has led to the development of technologies and strategies for the glycosylation engineering of novel antibiotics. Amino-, C-branched and O-methylated 6-deoxyhexoses play a favourite role in the biosynthesis of clinically important antibiotics like tylosin, erythromycin or oleandomycin and are essential for the antibiotic activity. They are transferred onto the aglycon by glycosyltransferases using dTDP-activated deoxyhexoses. The in vitro biochemical characterization of the biosynthetic enzymes and the glycosyltransferases are, however, hampered due to the poor synthetic access to dTDP-activated deoxysugars and their biosynthetic intermediates. The overcoming of the poor availability of dTDP-activated sugars was the target of several researchers to fulfil their distinct aims with these sugars which were mostly involved in the synthesis of different biological active compounds. Several completely different strategies were used in the past years to improve the availability of dTDP-activated deoxysugars, varying from complete enzymatic synthesis via syntheses using reaction technology for yield optimization to full organic synthesis or shortcuts like the decomposition of commercially available antibiotics and later chemical activation of the sugar moieties. This review gives a survey of the synthesis of dTDP-activated sugars by chemical and chemoenzymatic approaches and discusses the promiscuity of glycosyltransferases to evaluate the chances for applying them for the production of new bioactive compounds. It summarizes the most important enzymes in the field of synthesis using biosynthetic pathway enzymes and describes solutions for occurring challenges during application. Finally, this review will give a survey about the availability of dTDP-activated sugars in sufficient scale and will also point at important sugars which are still bottlenecks and difficult to synthesize and therefore should become a target for enhanced research efforts.

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Section: Chemo-enzymatic Synthesismentioning

confidence: 99%

Section: Chemo-enzymatic Synthesismentioning

confidence: 99%

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Nucleotide Deoxysugars: Essential Tools for the Glycosylation Engineering of Novel Bioactive Compounds

Rupprath¹,

Schumacher²,

Elling

2005

CMC

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“…Some examples include the efficient in vitro enzymatic syntheses of TDP-4-keto-6-deoxy-Dglucose (17) [164][165][166], TDP-L-mycarose (34) of the tylosin pathway [62], TDP-L-eremosamine (29) of the chloroeremomycin pathway [52], TDP-D-forosamine (43) of the spinosyn pathway [81], and TDP-L-digitoxose (36) of the kijanimicin pathway [66]. These syntheses were carried out in one-or two-pot reactions.…”

Section: Glycodiversification Via Enzyme Engineeringmentioning

confidence: 99%

Pathway and Enzyme Engineering and Applications for Glycodiversification

Zhao

Liu²

2010

Enzyme Technologies

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“…Due to such versatility, the ready availability of this sugar is important for the investigation of many interesting deoxysugar biosynthetic enzymes. Not surprisingly, several useful methods for the synthesis of 6 have been developed (Elling et al, 2005; Oh et al, 2003; Rupprath et al, 2005; Stein et al, 1998). …”

Section: Introductionmentioning

confidence: 99%

Chapter 21 Enzymatic Synthesis of TDP-Deoxysugars

White-Phillip

Thibodeaux

Liu

2009

Methods in Enzymology

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Many biologically active bacterial natural products contain highly modified deoxysugar residues that are often critical for the activity of the parent compounds. Most of these deoxysugars are secondary metabolites that are biosynthesized in the form of nucleotide diphosphate (NDP) sugars prior to their transfer to natural product aglycones by glycosyltransferases. Over the past decade, many biosynthetic pathways that lead to the formation of these unusual sugars have been unraveled, and the mechanisms of many key enzymatic transformations involved in these pathways have been elucidated. However, obtaining workable quantities of NDP-deoxysugars for in vitro studies is often a difficult task. This limitation has hindered an in-depth investigation of the substrate specificity of deoxysugar biosynthetic enzymes, many of which are promiscuous with respect to their NDP-sugar substrates and are, thus, potentially useful catalysts for natural product glycoengineering. Presented in this review are procedures for the enzymatic synthesis and purification of a variety of NDP-deoxysugars, including some early intermediates in NDP-deoxysugar biosynthetic pathways, and highly modified NDP-deoxysugars that are late intermediates in their respective biosynthetic pathways. The procedures described herein could be used as general guidelines for the development of specific protocols for the synthesis of other NDP-deoxysugars.

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An Enzyme Module System for the Synthesis of dTDP‐activated Deoxysugars from dTMP and Sucrose

Cited by 41 publications

References 72 publications

Nucleotide Deoxysugars: Essential Tools for the Glycosylation Engineering of Novel Bioactive Compounds

Nucleotide Deoxysugars: Essential Tools for the Glycosylation Engineering of Novel Bioactive Compounds

Pathway and Enzyme Engineering and Applications for Glycodiversification

Chapter 21 Enzymatic Synthesis of TDP-Deoxysugars

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