Characterizing non-hydrolyzing Neisseria meningitidis serogroup A UDP-N-acetylglucosamine (UDP-GlcNAc) 2-epimerase using UDP-N-acetylmannosamine (UDP-ManNAc) and derivatives

Zhang, Lei; Muthana, Musleh M.; Yu, Hai; McArthur, John B.; Qu, Jingyao; Chen, Xi

doi:10.1016/j.carres.2015.10.016

Cited by 12 publications

(18 citation statements)

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“…Sugar nucleotides and derivatives were enzymatically or chemoenzymatically synthesized previously. 38, 40, 52–55 …”

Section: Methodsmentioning

confidence: 99%

Donor substrate promiscuity of bacterial β1–3-N-acetylglucosaminyltransferases and acceptor substrate flexibility of β1–4-galactosyltransferases

Xue

et al. 2016

Bioorganic & Medicinal Chemistry

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β1–3-N-Acetylglucosaminyltransferases (β3GlcNAcTs) and β1–4-galactosyltransferases (β4GalTs) have been broadly used in enzymatic synthesis of N-acetyllactosamine (LacNAc)-containing oligosaccharides and glycoconjugates including poly-LacNAc, and lacto-N-neotetraose (LNnT) found in the milk of human and other mammals. In order to explore oligosaccharides and derivatives that can be synthesized by the combination of β3GlcNAcTs and β4GalTs, donor substrate specificity studies of two bacterial β3GlcNAcTs from Helicobacter pylori (Hpβ3GlcNAcT) and Neisseria meningitidis (NmLgtA), respectively, using a library of 39 sugar nucleotides were carried out. The two β3GlcNAcTs have complementary donor substrate promiscuity and 13 different trisaccharides were produced. They were used to investigate the acceptor substrate specificities of three β4GalTs from Neisseria meningitidis (NmLgtB), Helicobacter pylori (Hpβ4GalT), and bovine (Bβ4GalT), respectively. Ten of the 13 trisaccharides were shown to be tolerable acceptors for at least one of these β4GalTs. The application of NmLgtA in one-pot multienzyme (OPME) synthesis of two trisaccharides including GalNAcβ1–3Galβ1–4GlcβProN3 and Galβ1–3Galβ1–4Glc was demonstrated. The study provides important information for using these glycosyltransferases as powerful catalysts in enzymatic and chemoenzymatic syntheses of oligosaccharides and derivatives which can be useful probes and reagents.

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“…Sugar nucleotides and derivatives were enzymatically or chemoenzymatically synthesized previously. 38, 40, 52–55 …”

Section: Methodsmentioning

confidence: 99%

Donor substrate promiscuity of bacterial β1–3-N-acetylglucosaminyltransferases and acceptor substrate flexibility of β1–4-galactosyltransferases

Xue

et al. 2016

Bioorganic & Medicinal Chemistry

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“…UDP-GlcNAc was not required for the activity of NmSacA in epimerizing UDP-ManNAc to UDP-GlcNAc, but UDP-GlcNAc did appear to increase the initial rate of epimerization (Zhang et al, 2016). To better understand the epimerization reaction mechanism of NmSacA and its apparent lack of a requirement for UDP-GlcNAc for reactivity, and to determine the structural basis of its activity on modified substrates, we solved two crystal structures of NmSacA: a ligand-free structure and a structure bound to the ligand UDP-GlcNAc.…”

Section: Introductionmentioning

confidence: 86%

“…1; Morgan et al, 1997;Tanner, 2002). NmSacA was shown to release 2-acetamidoglucal and UDP, which were easily observed when the reactions were carried out for an extended period of time (Zhang et al, 2016).…”

Section: Introductionmentioning

confidence: 98%

“…The cps operon of N. meningitidis serogroup A contains four open reading frames, with the first being sacA, which encodes an enzyme (NmSacA) that catalyzes the first step in the N. meningitidis serogroup A CPS biosynthetic pathway (Swartley et al, 1998). This enzyme (EC 5.1.3.14) catalyzes the interconversion of uridine 5 0 -diphosphate N-acetylglucosamine (UDP-GlcNAc) to its C2 00 -epimer UDP-N-acetylmannosamine (UDP-ManNAc) (Zhang et al, 2016;Swartley et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

“…NmSacA has been verified to be a nonhydrolyzing UDP-GlcNAc 2-epimerase (Zhang et al, 2016). The mechanism of this type of bacterial epimerase is believed to involve anti elimination of the C2 00 proton and UDP from UDP-GlcNAc, generating enzyme-bound intermediates of UDP and 2-acetamidoglucal, followed by the subsequent syn addition of a proton to C2 00 and UDP to the same face of the double bond, producing the UDP-ManNAc product ( Fig.…”

Section: Introductionmentioning

confidence: 99%

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Structural characterization of a nonhydrolyzing UDP-GlcNAc 2-epimerase fromNeisseria meningitidisserogroup A

et al. 2020

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Bacterial nonhydrolyzing UDP-N-acetylglucosamine 2-epimerases catalyze the reversible interconversion of UDP-N-acetylglucosamine (UDP-GlcNAc) and UDP-N-acetylmannosamine (UDP-ManNAc). UDP-ManNAc is an important intermediate in the biosynthesis of certain cell-surface polysaccharides, including those in some pathogenic bacteria, such as Neisseria meningitidis and Streptococcus pneumoniae. Many of these epimerases are allosterically regulated by UDP-GlcNAc, which binds adjacent to the active site and is required to initiate UDP-ManNAc epimerization. Here, two crystal structures of UDP-N-acetylglucosamine 2-epimerase from Neisseria meningitidis serogroup A (NmSacA) are presented. One crystal structure is of the substrate-free enzyme, while the other structure contains UDP-GlcNAc substrate bound to the active site. Both structures form dimers as seen in similar epimerases, and substrate binding to the active site induces a large conformational change in which two Rossmann-like domains clamp down on the substrate. Unlike other epimerases, NmSacA does not require UDP-GlcNAc to instigate the epimerization of UDP-ManNAc, although UDP-GlcNAc was found to enhance the rate of epimerization. In spite of the conservation of residues involved in binding the allosteric UDP-GlcNAc observed in similar UDP-GlcNAc 2-epimerases, the structures presented here do not contain UDP-GlcNAc bound in the allosteric site. These structural results provide additional insight into the mechanism and regulation of this critical enzyme and improve the structural understanding of the ability of NmSacA to epimerize modified substrates.

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Stereoselective Thioconjugation by Photoinduced Thiol‐ene Coupling Reactions of Hexo‐ and Pentopyranosyl d‐ and l‐Glycals at Low‐Temperature—Reactivity and Stereoselectivity Study

Kelemen

Bege

Eszenyi

et al. 2019

Chemistry A European J

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A comprehensive optimization and mechanistic study on the photoinduced hydrothiolation of different d‐ and l‐ hexo‐ and pentoglycals with various thiols was performed, at the temperature range of RT to −120 °C. Addition of thiols onto 2‐substituted hexoglycals proceeded with complete 1,2‐cis‐α‐stereoselectivity in all cases. Hydrothiolation of 2‐substituted pentoglycals resulted in mixtures of 1,2‐cis‐α‐ and ‐β‐thioglycosides of varying ratio depending on the configuration of the reactants. Hydrothiolation of unsubstituted glycals at −80 °C proceeded with excellent yields and, except for galactal, provided the axially C2‐S‐linked isomers with high selectivity. Cooling was always beneficial to the efficacy, increased the yields and in most cases significantly raised the stereoselectivity. The suggested mechanism explains the different conformational preferences of the intermediate carbon‐centered radicals, which is a crucial factor in the stereoselectivity of the reactions.

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Characterizing non-hydrolyzing Neisseria meningitidis serogroup A UDP-N-acetylglucosamine (UDP-GlcNAc) 2-epimerase using UDP-N-acetylmannosamine (UDP-ManNAc) and derivatives

Cited by 12 publications

References 70 publications

Donor substrate promiscuity of bacterial β1–3-N-acetylglucosaminyltransferases and acceptor substrate flexibility of β1–4-galactosyltransferases

Donor substrate promiscuity of bacterial β1–3-N-acetylglucosaminyltransferases and acceptor substrate flexibility of β1–4-galactosyltransferases

Structural characterization of a nonhydrolyzing UDP-GlcNAc 2-epimerase fromNeisseria meningitidisserogroup A

Stereoselective Thioconjugation by Photoinduced Thiol‐ene Coupling Reactions of Hexo‐ and Pentopyranosyl d‐ and l‐Glycals at Low‐Temperature—Reactivity and Stereoselectivity Study

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