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Literature summary for 5.1.3.14 extracted from

  • Zhang, L.; Muthana, M.M.; Yu, H.; McArthur, J.B.; Qu, J.; Chen, X.
    Characterizing non-hydrolyzing Neisseria meningitidis serogroup A UDP-N-acetylglucosamine (UDP-GlcNAc) 2-epimerase using UDP-N-acetylmannosamine (UDP-ManNAc) and derivatives (2016), Carbohydr. Res., 419, 18-28 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.1.3.14

Activating Compound

Activating Compound Comment Organism Structure
additional information no activation by EDTA, and 1 mM DTT Neisseria meningitidis serogroup A / serotype 4A
UDP-GlcNAc allosteric activator, absolutely required for their UDP-ManNAc 2-epimerization catalytic activity, activates recombinant His6-tagged enzyme NmSacA Neisseria meningitidis serogroup A / serotype 4A
UDP-N-acetyl-alpha-D-glucosamine activates the recombinant enzyme, UDP-GlcNAc is able to activate the epimerization process in a dose-dependent manner Neisseria meningitidis serogroup A /serotype 4A

Cloned(Commentary)

Cloned (Comment) Organism
gene sacA, encoded in the serogroup A cps operon, sequence comparisons, recombinant overexpression of C-terminally His6-tagged recombinant NmSacA in Escherichia coli strain BL21(DE3) Neisseria meningitidis serogroup A / serotype 4A
gene sacA, recombinant expression of C-terminally His6-tagged enzyme NmSacA in Escherichia coli strain BL21(DE3) Neisseria meningitidis serogroup A /serotype 4A

Inhibitors

Inhibitors Comment Organism Structure
2-acetamidoglucal
-
 Neisseria meningitidis serogroup A / serotype 4A
2-acetamidoglucal recombinant enzyme Neisseria meningitidis serogroup A /serotype 4A
Ca2+
-
 Neisseria meningitidis serogroup A / serotype 4A
Ca2+ slight inhibition at 10 mM Neisseria meningitidis serogroup A /serotype 4A
Co2+ strong inhibition at 10 mM Neisseria meningitidis serogroup A /serotype 4A
Cu2+
-
 Neisseria meningitidis serogroup A / serotype 4A
Cu2+ slight inhibition at 10 mM Neisseria meningitidis serogroup A /serotype 4A
Mg2+
-
 Neisseria meningitidis serogroup A / serotype 4A
Mg2+ slight inhibition at 10 mM Neisseria meningitidis serogroup A /serotype 4A
Mn2+ complete inhibition at 10 mM, the epimerase and the activity cannot be rescued by subsequent addition of EDTA Neisseria meningitidis serogroup A /serotype 4A
Mn2+ leads to enzyme precipitation at 10 mM Neisseria meningitidis serogroup A / serotype 4A
additional information both 2-acetamidoglucal and UDP show competitive inhibitory effects for the UDP-ManNAc 2-epimerization of NmSacA-His6. No inhibition by EDTA, and 1 mM DTT Neisseria meningitidis serogroup A / serotype 4A
Ni2+
-
 Neisseria meningitidis serogroup A / serotype 4A
Ni2+ slight inhibition at 10 mM Neisseria meningitidis serogroup A /serotype 4A
UDP
-
 Neisseria meningitidis serogroup A / serotype 4A
UDP recombinant enzyme Neisseria meningitidis serogroup A /serotype 4A
Zn2+ complete inhibition at 10 mM, the epimerase and the activity cannot be rescued by subsequent addition of EDTA Neisseria meningitidis serogroup A /serotype 4A
Zn2+ leads to enzyme precipitation at 10 mM Neisseria meningitidis serogroup A / serotype 4A

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.6
-
 UDP-N-acetyl-alpha-D-glucosamine pH 8.5, 37°C, recombinant enzyme Neisseria meningitidis serogroup A / serotype 4A
5.2
-
 UDP-N-acetyl-alpha-D-mannosamine pH 8.5, 37°C, recombinant enzyme Neisseria meningitidis serogroup A / serotype 4A

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the 2-epimerase activity of NmSacA-His6 is not dependent on the addition of an external divalent metal ion, Li+ and EDTA have no effect on enzyme activity Neisseria meningitidis serogroup A / serotype 4A
additional information the 2-epimerase activity of NmSacA-His6 is not dependent on the addition of an external divalent metal ion. No effect by 10 mM of EDTA and Li+, poor effect by 1 mM DTT Neisseria meningitidis serogroup A /serotype 4A

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-N-acetyl-alpha-D-glucosamine Neisseria meningitidis serogroup A / serotype 4A
-
 UDP-N-acetyl-alpha-D-mannosamine
-
 r
UDP-N-acetyl-alpha-D-glucosamine Neisseria meningitidis serogroup A /serotype 4A
-
 UDP-N-acetyl-alpha-D-mannosamine
-
 r
UDP-N-acetyl-alpha-D-glucosamine Neisseria meningitidis serogroup A / serotype 4A Z2491
-
 UDP-N-acetyl-alpha-D-mannosamine
-
 r
UDP-N-acetyl-alpha-D-glucosamine Neisseria meningitidis serogroup A /serotype 4A Z2491
-
 UDP-N-acetyl-alpha-D-mannosamine
-
 r

Organism

Organism UniProt Comment Textmining
Neisseria meningitidis serogroup A / serotype 4A A0A0U1RGY0
-
-
Neisseria meningitidis serogroup A / serotype 4A Z2491 A0A0U1RGY0
-
-
Neisseria meningitidis serogroup A /serotype 4A A0A0U1RGY0
-
-
Neisseria meningitidis serogroup A /serotype 4A Z2491 A0A0U1RGY0
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant C-terminally His6-tagged enzyme NmSacA from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Neisseria meningitidis serogroup A /serotype 4A
recombinant C-terminally His6-tagged NmSacA from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Neisseria meningitidis serogroup A / serotype 4A

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dihydropyrimidine-2,4(1H,3H)-dione-alpha-D-ribofuranosyl-5'-diphospho-2-acetamido-2-deoxy-alpha-D-mannopyranoside
-
 Neisseria meningitidis serogroup A / serotype 4A ?
-
 ?
dihydropyrimidine-2,4(1H,3H)-dione-alpha-D-ribofuranosyl-5'-diphospho-2-acetamido-2-deoxy-alpha-D-mannopyranoside
-
 Neisseria meningitidis serogroup A / serotype 4A Z2491 ?
-
 ?
additional information substrate specificity, overview. Recombinant His6-tagged NmSacA-His6 can tolerate several chemoenzymatically synthesized UDP-ManNAc derivatives as substrates in the absence of UDP-GlcNAc although its activity is much lower than with non-modified UDP-ManNAc. Homology modeling and molecular docking. The formation of UDP-GlcNAc or UDP-ManNAc is not observed by incubating NmSacA-His6 with 2-acetamidoglucal and UDP in contrast to the serogroup B enzyme. No enzyme activity with UDP-mannose, UDP-ManF, UDP-ManN3, UDP-ManNH2, uridine 5'-diphospho-2-butyramido-2-deoxy-alpha-D-mannopyranoside, uridine 5'-diphospho-2-azidoacetamido-2-deoxy-alpha-D-mannopyranoside, uridine 5'-diphospho-2-phenylacetamido-2-deoxy-alpha-D-mannopyranoside, dihydropyrimidine-2, 4(1H,3H)-dione-alpha-D-ribofuranosyl-5'-diphospho-2-azidoxyacetamido-2-deoxy-alpha-D-mannopyranoside, and uridine 5'-diphospho-2-azidoacetamido-2-deoxy-alpha-D-glucopyranoside, docking study Neisseria meningitidis serogroup A / serotype 4A ?
-
 ?
additional information the recombinnat enzyme His6-tagged NmSacA tolerates several chemoenzymatically synthesized UDP-ManNAc derivatives as substrates although its activity is much lower than with non-modified UDP-ManNAc, substrate specificity, overview Neisseria meningitidis serogroup A /serotype 4A ?
-
 ?
additional information substrate specificity, overview. Recombinant His6-tagged NmSacA-His6 can tolerate several chemoenzymatically synthesized UDP-ManNAc derivatives as substrates in the absence of UDP-GlcNAc although its activity is much lower than with non-modified UDP-ManNAc. Homology modeling and molecular docking. The formation of UDP-GlcNAc or UDP-ManNAc is not observed by incubating NmSacA-His6 with 2-acetamidoglucal and UDP in contrast to the serogroup B enzyme. No enzyme activity with UDP-mannose, UDP-ManF, UDP-ManN3, UDP-ManNH2, uridine 5'-diphospho-2-butyramido-2-deoxy-alpha-D-mannopyranoside, uridine 5'-diphospho-2-azidoacetamido-2-deoxy-alpha-D-mannopyranoside, uridine 5'-diphospho-2-phenylacetamido-2-deoxy-alpha-D-mannopyranoside, dihydropyrimidine-2, 4(1H,3H)-dione-alpha-D-ribofuranosyl-5'-diphospho-2-azidoxyacetamido-2-deoxy-alpha-D-mannopyranoside, and uridine 5'-diphospho-2-azidoacetamido-2-deoxy-alpha-D-glucopyranoside, docking study Neisseria meningitidis serogroup A / serotype 4A Z2491 ?
-
 ?
additional information the recombinnat enzyme His6-tagged NmSacA tolerates several chemoenzymatically synthesized UDP-ManNAc derivatives as substrates although its activity is much lower than with non-modified UDP-ManNAc, substrate specificity, overview Neisseria meningitidis serogroup A /serotype 4A Z2491 ?
-
 ?
UDP-N-acetyl-alpha-D-glucosamine
-
 Neisseria meningitidis serogroup A / serotype 4A UDP-N-acetyl-alpha-D-mannosamine
-
 r
UDP-N-acetyl-alpha-D-glucosamine
-
 Neisseria meningitidis serogroup A /serotype 4A UDP-N-acetyl-alpha-D-mannosamine
-
 r
UDP-N-acetyl-alpha-D-glucosamine interconversion Neisseria meningitidis serogroup A /serotype 4A UDP-N-acetyl-alpha-D-mannosamine
-
 r
UDP-N-acetyl-alpha-D-glucosamine
-
 Neisseria meningitidis serogroup A / serotype 4A Z2491 UDP-N-acetyl-alpha-D-mannosamine
-
 r
UDP-N-acetyl-alpha-D-glucosamine
-
 Neisseria meningitidis serogroup A /serotype 4A Z2491 UDP-N-acetyl-alpha-D-mannosamine
-
 r
UDP-N-acetyl-alpha-D-glucosamine interconversion Neisseria meningitidis serogroup A /serotype 4A Z2491 UDP-N-acetyl-alpha-D-mannosamine
-
 r
UDP-N-acetyl-alpha-D-mannosamine
-
 Neisseria meningitidis serogroup A / serotype 4A UDP-N-acetyl-alpha-D-glucosamine
-
 r
UDP-N-acetyl-alpha-D-mannosamine interconversion Neisseria meningitidis serogroup A /serotype 4A UDP-N-acetyl-alpha-D-glucosamine
-
 r
UDP-N-acetyl-alpha-D-mannosamine
-
 Neisseria meningitidis serogroup A / serotype 4A Z2491 UDP-N-acetyl-alpha-D-glucosamine
-
 r
UDP-N-acetyl-alpha-D-mannosamine interconversion Neisseria meningitidis serogroup A /serotype 4A Z2491 UDP-N-acetyl-alpha-D-glucosamine
-
 r
uridine 5'-diphospho-2-hydroxyacetamido-2-deoxy-alpha-D-mannopyranoside
-
 Neisseria meningitidis serogroup A / serotype 4A ?
-
 ?
uridine 5'-diphospho-2-hydroxyacetamido-2-deoxy-alpha-D-mannopyranoside
-
 Neisseria meningitidis serogroup A / serotype 4A Z2491 ?
-
 ?
uridine 5'-diphospho-2-propionamido-2-deoxy-alpha-D-mannopyranoside
-
 Neisseria meningitidis serogroup A / serotype 4A ?
-
 ?
uridine 5'-diphospho-N-trifluoroaceto-2-deoxy-alpha-D-mannopyranoside
-
 Neisseria meningitidis serogroup A / serotype 4A ?
-
 ?

Subunits

Subunits Comment Organism
? x * 43000, recombinant His6-tagged enzyme, SDS-PAGE Neisseria meningitidis serogroup A /serotype 4A
? x * 43000, recombinant His6-tagged NmSacA, SDS-PAGE Neisseria meningitidis serogroup A / serotype 4A

Synonyms

Synonyms Comment Organism
NmSacA
-
 Neisseria meningitidis serogroup A / serotype 4A
NmSacA
-
 Neisseria meningitidis serogroup A /serotype 4A
non-hydrolyzing Neisseria meningitidis serogroup A UDP-GlcNAc 2-epimerase
-
 Neisseria meningitidis serogroup A /serotype 4A
non-hydrolyzing Neisseria meningitidis serogroup A UDP-N-acetylglucosamine 2-epimerase
-
 Neisseria meningitidis serogroup A /serotype 4A
non-hydrolyzing UDP-GlcNAc 2-epimerase
-
 Neisseria meningitidis serogroup A / serotype 4A
non-hydrolyzing UDP-GlcNAc 2-epimerase
-
 Neisseria meningitidis serogroup A /serotype 4A
non-hydrolyzing UDP-N-acetylglucosamine 2-epimerase
-
 Neisseria meningitidis serogroup A / serotype 4A
non-hydrolyzing uridine 5'-diphosphate-N-acetylglucosamine 2-epimerase
-
 Neisseria meningitidis serogroup A / serotype 4A
SacA
-
 Neisseria meningitidis serogroup A /serotype 4A
SacA
-
 Neisseria meningitidis serogroup A / serotype 4A
UDP-GlcNAc 2-epimerase
-
 Neisseria meningitidis serogroup A / serotype 4A

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Neisseria meningitidis serogroup A / serotype 4A

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
31
-
 UDP-N-acetyl-alpha-D-glucosamine pH 8.5, 37°C, recombinant enzyme Neisseria meningitidis serogroup A / serotype 4A
52
-
 UDP-N-acetyl-alpha-D-mannosamine pH 8.5, 37°C, recombinant enzyme Neisseria meningitidis serogroup A / serotype 4A

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8 9
-
 Neisseria meningitidis serogroup A / serotype 4A
8 9 recombinant enzyme Neisseria meningitidis serogroup A /serotype 4A

pH Range

pH Minimum pH Maximum Comment Organism
6 11 activity range, recombinant enzyme Neisseria meningitidis serogroup A /serotype 4A
6 11 activity range with high activity Neisseria meningitidis serogroup A / serotype 4A

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.8
-
 UDP pH 8.5, 37°C, recombinant enzyme Neisseria meningitidis serogroup A / serotype 4A
14
-
 2-acetamidoglucal pH 8.5, 37°C, recombinant enzyme Neisseria meningitidis serogroup A / serotype 4A

General Information

General Information Comment Organism
additional information of the six major disease-causing Neisseria meningitidis serogroups, only serogroups A and X produce capsular polysaccharides (CPSs) that do not have N-acetylneuraminic acid (Neu5Ac, sialic acid is a more general term) residues. Unlike the CPSs of serogroups B and C which are homopolymers of Neu5Ac with alpha2-8- and alpha2-9-linkages respectively, or serogroups W-135 and Y CPSs which are heteropolymers of [-6Gal/Glcalpha1-4Neu5Acalpha2-]n with alternating Neu5Ac and Gal/Glc as disaccharide repeating units, the CPS of serogroup A is a homopolymer of [-6ManNAcalpha1-phosphate-]n. Correspondingly, the genetic organization for serogroup A capsule is different from those for serogroups B, C, W-135, and Y Neisseria meningitidis serogroup A / serotype 4A
additional information homology modeling and molecular docking reveal structural determinants of NmSacA substrate specificity. Disulfide bond formation is not required for the epimerase activity of NmSacA-His6 Neisseria meningitidis serogroup A /serotype 4A
physiological function SacA is proposed to be involved in the first step in serogroup A capsular polysaccharides (CPS) biosynthetic pathway and is critical for serogroup A CPS biosynthesis. It catalyzes the interconversion between UDP-GlcNAc and UDP-ManNAc. This epimerization process is independent of nicotinamide adenine dinucleotide oxidized form (NAD+) and is critical for synthesizing bacterial ManNAc-containing CPSs Neisseria meningitidis serogroup A / serotype 4A
physiological function Neisseria meningitidis serogroup A non-hydrolyzing UDP-GlcNAc 2-epimerase (NmSacA) catalyzes the interconversion between UDP-GlcNAc and uridine 5?-diphosphate-N-acetylmannosamine (UDP-ManNAc). It is a key enzyme involved in the biosynthesis of the capsular polysaccharide [-6ManNAc?1-phosphate-]n of N. meningitidis serogroup A, one of the six serogroups (A, B, C, W-135, X, and Y) that account for most cases of Neisseria meningitidis-caused bacterial septicemia and meningitis Neisseria meningitidis serogroup A /serotype 4A

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
8.6
-
 UDP-N-acetyl-alpha-D-mannosamine pH 8.5, 37°C, recombinant enzyme Neisseria meningitidis serogroup A / serotype 4A
10
-
 UDP-N-acetyl-alpha-D-glucosamine pH 8.5, 37°C, recombinant enzyme Neisseria meningitidis serogroup A / serotype 4A