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Literature summary extracted from

  • Nishimoto, M.; Kitaoka, M.
    Identification of N-acetylhexosamine 1-kinase in the complete lacto-N-biose I/galacto-N-biose metabolic pathway in Bifidobacterium longum (2007), Appl. Environ. Microbiol., 73, 6444-6449.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.1.162 isopropyl-1-thio-beta-D-galactoside induction of protein expression at a final concentration of 0.5 mM Bifidobacterium longum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.162 Escherichia coli strains select96 and BL21(DE3) are used as hosts for cloning and expression. Bifidobacterium longum
2.7.1.162 expression in Escherichia coli Bifidobacterium longum

General Stability

EC Number General Stability Organism
2.7.1.162 loses half of enzyme activity during incubation for 30 min at 50°C in 0.1 M Tris buffer (pH 8.5) Bifidobacterium longum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.162 0.065
-
 N-acetyl-alpha-D-galactosamine
-
 Bifidobacterium longum
2.7.1.162 0.065
-
 N-acetyl-alpha-D-galactosamine pH 8.5, 30°C Bifidobacterium longum
2.7.1.162 0.118
-
 N-acetyl-alpha-D-glucosamine
-
 Bifidobacterium longum
2.7.1.162 0.118
-
 N-acetyl-alpha-D-glucosamine pH 8.5, 30°C Bifidobacterium longum
2.7.1.162 0.172
-
 ATP cosubstrate: N-acetyl-alpha-D-glucosamine, pH 8.5, 30°C Bifidobacterium longum
2.7.1.162 0.172
-
 ATP at N-acetyl-alpha-D-glucosamine saturation Bifidobacterium longum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.162 Ca2+ 1 mM, 1% relative activity compared to Mg2+ Bifidobacterium longum
2.7.1.162 Co2+ 21% of the activity with Mg2+ Bifidobacterium longum
2.7.1.162 Co2+ 1 mM, 21% relative activity compared to Mg2+ Bifidobacterium longum
2.7.1.162 Mg2+ requires a divalent cation, with Mg2+ resulting in the greatest stimulation of enzyme activity Bifidobacterium longum
2.7.1.162 Mg2+ 1 mM, 100% relative activity Bifidobacterium longum
2.7.1.162 Mn2+ 1 mM, 7% relative activity compared to Mg2+ Bifidobacterium longum
2.7.1.162 Ni2+ 1 mM, 4% relative activity compared to Mg2+ Bifidobacterium longum
2.7.1.162 Zn2+ 1 mM, 8% relative activity compared to Mg2+ Bifidobacterium longum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.162 ATP + N-acetyl-alpha-D-galactosamine Bifidobacterium longum the enzyme is involved in the lacto-N-biose I/galacto-N-biose degradation pathway ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-galactosamine Bifidobacterium longum lacto-N-biose I/galacto-N-biose metabolic pathway ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-galactosamine Bifidobacterium longum JCM 1217 the enzyme is involved in the lacto-N-biose I/galacto-N-biose degradation pathway ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-galactosamine Bifidobacterium longum JCM 1217 lacto-N-biose I/galacto-N-biose metabolic pathway ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-glucosamine Bifidobacterium longum
-
 ADP + N-acetyl-alpha-D-glucosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-glucosamine Bifidobacterium longum the enzyme is involved in the lacto-N-biose I/galacto-N-biose degradation pathway ADP + N-acetyl-alpha-D-glucosamine 1-phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.211 Bifidobacterium longum subsp. Longum
-
-
-
2.4.1.211 Bifidobacterium longum subsp. Longum JCM 1217
-
-
-
2.7.1.162 Bifidobacterium longum E8MF12 strain JCM1217
-
2.7.1.162 Bifidobacterium longum JCM 1217 E8MF12 strain JCM1217
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.162
-
 Bifidobacterium longum
2.7.1.162 The transformant is cultivated and protein expression is induced by the addition of isopropyl-1-thio-beta-D-galactoside. The cells are harvested by centrifugation and sonicated. Cell debris is removed by centrifugation and enzyme is purified on an Ni-nitrilotriacetic acid agarose gel. Bifidobacterium longum

Reaction

EC Number Reaction Comment Organism Reaction ID
2.7.1.162 ATP + N-acetyl-D-hexosamine = ADP + N-acetyl-alpha-D-hexosamine 1-phosphate enzyme acts by a sequential bi bi (two substrates-two products) mechanism, with the reaction occurring after the binding of both ATP and N-acetylhexosamine Bifidobacterium longum
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.1.162 cell culture strain JCM1217 Bifidobacterium longum
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.162 additional information
-
 Incubation of 4 microM LnpB protein with 10 mM N-acetyl-alpha-D-glucosamine 6-sulfate in 0.1 M MOPS buffer (pH 7.5) at 30°C for 30 min shows that the protein have no hydrolytic activity Bifidobacterium longum
2.7.1.162 0.001
-
 D-glucose, substrate donor specificity Bifidobacterium longum
2.7.1.162 0.002
-
 D-mannosamine, substrate donor specificity Bifidobacterium longum
2.7.1.162 0.004
-
 D-glucosamine, substrate donor specificity Bifidobacterium longum
2.7.1.162 0.007
-
 2-deoxyglucose, substrate donor specificity Bifidobacterium longum
2.7.1.162 0.024
-
 D-mannose, substrate donor specificity Bifidobacterium longum
2.7.1.162 0.049
-
 D-talose, substrate donor specificity Bifidobacterium longum
2.7.1.162 0.22
-
 N-acetyl-D-mannosamine, substrate donor specificity Bifidobacterium longum
2.7.1.162 0.48
-
 ITP, substrate donor specificity Bifidobacterium longum
2.7.1.162 0.65
-
 GTP, substrate donor specificity Bifidobacterium longum
2.7.1.162 0.89
-
 N-acetyl-D-galactosamine, substrate donor specificity Bifidobacterium longum
2.7.1.162 1.48
-
 ATP, substrate donor specificity Bifidobacterium longum
2.7.1.162 1.49
-
 N-acetyl-D-glucosamine, substrate donor specificity Bifidobacterium longum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.211 beta-D-galactopyranosyl-1,3-N-acetyl-D-galactosmine + phosphate the gene operon for LNBP lnpABCD encodes a lacto-N-biose I and beta-D-galactopyranosyl-1,3-N-acetyl-D-galactosmine metabolic pathway with lacto-N-biose phosphorylase, N-acetylhexosamine 1-kinase, UDP-glucose hexose 1-phosphate uridylyltransferase, and UDP-glucose 4-epimerase, this pathway is involved in the intestinal colonization of bifidobacteria and utilizes lacto-N-biose I from human milk oligosaccharides or beta-D-galactopyranosyl-1,3-N-acetyl-D-galactosmine from mucin sugars Bifidobacterium longum subsp. Longum alpha-D-galactose 1-phosphate + N-acetyl-D-galactosamine
-
 ?
2.4.1.211 beta-D-galactopyranosyl-1,3-N-acetyl-D-galactosmine + phosphate the gene operon for LNBP lnpABCD encodes a lacto-N-biose I and beta-D-galactopyranosyl-1,3-N-acetyl-D-galactosmine metabolic pathway with lacto-N-biose phosphorylase, N-acetylhexosamine 1-kinase, UDP-glucose hexose 1-phosphate uridylyltransferase, and UDP-glucose 4-epimerase, this pathway is involved in the intestinal colonization of bifidobacteria and utilizes lacto-N-biose I from human milk oligosaccharides or beta-D-galactopyranosyl-1,3-N-acetyl-D-galactosmine from mucin sugars Bifidobacterium longum subsp. Longum JCM 1217 alpha-D-galactose 1-phosphate + N-acetyl-D-galactosamine
-
 ?
2.4.1.211 beta-D-galactopyranosyl-1,3-N-acetyl-D-glucosamine + phosphate
-
 Bifidobacterium longum subsp. Longum alpha-D-galactose 1-phosphate + N-acetyl-D-glucosamine
-
 ?
2.4.1.211 beta-D-galactopyranosyl-1,3-N-acetyl-D-glucosamine + phosphate
-
 Bifidobacterium longum subsp. Longum JCM 1217 alpha-D-galactose 1-phosphate + N-acetyl-D-glucosamine
-
 ?
2.7.1.162 ATP + D-mannose phosphorylation at 1.6% of the activity with N-acetylglucosamine Bifidobacterium longum ?
-
 ?
2.7.1.162 ATP + D-mannose phosphorylation at 1.6% of the activity with N-acetylglucosamine Bifidobacterium longum JCM 1217 ?
-
 ?
2.7.1.162 ATP + D-talose phosphorylation at 3% of the activity with N-acetylglucosamine Bifidobacterium longum ?
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-galactosamine the enzyme is involved in the lacto-N-biose I/galacto-N-biose degradation pathway Bifidobacterium longum ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-galactosamine phosphorylation at 60% of the activity with N-acetylglucosamine Bifidobacterium longum ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-galactosamine lacto-N-biose I/galacto-N-biose metabolic pathway Bifidobacterium longum ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-galactosamine the enzyme is involved in the lacto-N-biose I/galacto-N-biose degradation pathway Bifidobacterium longum JCM 1217 ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-galactosamine phosphorylation at 60% of the activity with N-acetylglucosamine Bifidobacterium longum JCM 1217 ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-galactosamine lacto-N-biose I/galacto-N-biose metabolic pathway Bifidobacterium longum JCM 1217 ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-glucosamine
-
 Bifidobacterium longum ADP + N-acetyl-alpha-D-glucosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-alpha-D-glucosamine the enzyme is involved in the lacto-N-biose I/galacto-N-biose degradation pathway Bifidobacterium longum ADP + N-acetyl-alpha-D-glucosamine 1-phosphate
-
 ?
2.7.1.162 ATP + N-acetyl-D-mannosamine phosphorylation at 15% of the activity with N-acetylglucosamine Bifidobacterium longum ?
-
 ?
2.7.1.162 ATP + N-acetyl-D-mannosamine
-
 Bifidobacterium longum ADP + ?
-
 ?
2.7.1.162 GTP + N-acetyl-alpha-D-glucosamine GTP shows about 45% of the activity with ATP Bifidobacterium longum GDP + N-acetyl-alpha-D-glucosamine 1-phosphate
-
 ?
2.7.1.162 ITP + N-acetyl-alpha-D-glucosamine ITP shows about 30% of the activity with ATP Bifidobacterium longum IDP + N-acetyl-alpha-D-glucosamine 1-phosphate
-
 ?
2.7.1.162 additional information slightly active with D-talose, D-mannose, GTP, ITP Bifidobacterium longum ?
-
 ?
2.7.1.162 additional information slightly active with D-talose, D-mannose, GTP, ITP Bifidobacterium longum JCM 1217 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.211 1,3-beta-galactosyl-N-acetylhexosamine phosphorylase
-
 Bifidobacterium longum subsp. Longum
2.4.1.211 lacto-N-biose phosphorylase
-
 Bifidobacterium longum subsp. Longum
2.4.1.211 lnba
-
 Bifidobacterium longum subsp. Longum
2.4.1.211 LNBP
-
 Bifidobacterium longum subsp. Longum
2.7.1.162 lnpB
-
 Bifidobacterium longum
2.7.1.162 lnpB protein Bifidobacterium longum
2.7.1.162 N-acetylhexosamine 1-kinase
-
 Bifidobacterium longum
2.7.1.162 NahK
-
 Bifidobacterium longum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.162 40
-
-
 Bifidobacterium longum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.1.162 50
-
 pH 8.5, 30 min, 50% loss of activity Bifidobacterium longum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.162 0.752
-
 N-acetyl-alpha-D-galactosamine
-
 Bifidobacterium longum
2.7.1.162 1.21
-
 N-acetyl-alpha-D-glucosamine
-
 Bifidobacterium longum
2.7.1.162 1.21
-
 N-acetyl-alpha-D-glucosamine pH 8.5, 30°C Bifidobacterium longum
2.7.1.162 6.08
-
 N-acetyl-alpha-D-galactosamine pH 8.5, 30°C Bifidobacterium longum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.162 8.5
-
-
 Bifidobacterium longum
2.7.1.162 8.5
-
 assay at Bifidobacterium longum

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.7.1.162 5 9.5 stable Bifidobacterium longum
2.7.1.162 5 9.5 stability at 30°C Bifidobacterium longum

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.162 ATP
-
 Bifidobacterium longum
2.7.1.162 GTP
-
 Bifidobacterium longum
2.7.1.162 ITP
-
 Bifidobacterium longum