Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.4.1.212 extracted from

  • Eisele, A.; Zaun, H.; Kuballa, J.; Elling, L.
    In vitro one-pot enzymatic synthesis of hyaluronic acid from sucrose and N-acetylglucosamine optimization of the enzyme module system and nucleotide sugar regeneration (2018), ChemCatChem, 10, 2969-2981 .
No PubMed abstract available
Search for more literature for 2.4.1.212

Application

Application Comment Organism
synthesis multi-enzyme strategy for the in vitro one-pot synthesis of high-molecular-weight hyaluronic acid from substrates sucrose and N-acetylglucosamine (GlcNAc) with in situ regeneration of nucleotide sugars. With optimized reaction conditions, hyaluronic acid with a molecular mass above 2 MDa is synthesized from catalytic UDP concentrations and 10 mM GlcNAc in less than 10 h Pasteurella multocida

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-taggged residues 1-703 in Escherichia coli Pasteurella multocida

Inhibitors

Inhibitors Comment Organism Structure
UDP-glucuronate The substrate UDP-GlcA inhibits pmHAS in concentrations above 8 mM Pasteurella multocida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.8
-
 UDP-alpha-D-glucuronate pH 7.0, 25 °C, 10 mM MnCl2 and 15 mM UDP-GlcNAc (in absence of HA oligosaccharide as acceptor substrate) Pasteurella multocida
23.4
-
 UDP-N-acetyl-alpha-D-glucosamine pH 7.0, 25 °C, 10 mM MnCl2 and 5 mM UDP-GlcA (in absence of HA oligosaccharide as acceptor substrate) Pasteurella multocida

Metals/Ions

Metals/Ions Comment Organism Structure
K+ coactivation occurs if K+ is applied together with Mn2+ or Mg2+. With Mn2+, a high UDP reaction rate and high molecular mass of hyaluronic acid (2.45 MDa after 8 h) are achieved. With Mg2+, a low reaction rate and low molecular mass (1.55 MDa after 8 h) are reached. If 10 mM K+ are added to 15 mM Mg2+, a significant increase of the reaction rate by a factor of 2.7 is observed, and the molecular mass is doubled (3.11 MDa after 8 h). The coactivating effect of K+ together with Mn2+ is less pronounced Pasteurella multocida
Mg2+ Mg2+ activates with an optimum of 15-20 mM leading to about half of the yield obtained with Mn2+ after 8 h Pasteurella multocida
Mn2+ Mn2+ is the preferred metal ion with an optimum of 10 mM. Presence of Mn2+ induces a concentration-dependent decomposition of UDP-GlcA to its 1,2-cyclic phosphate and UMP, especially promoted at basic pH Pasteurella multocida

Organism

Organism UniProt Comment Textmining
Pasteurella multocida Q7BLV3
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.05
-
 presence of 10 mM Mn2+, pH 7.0, 25°C Pasteurella multocida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
[hyaluronan](n) + UDP-alpha-D-glucuronate
-
 Pasteurella multocida H+ + beta-D-glucuronosyl-(1->4)-[hyaluronan](n) + UDP
-
 ?
[hyaluronan](n) + UDP-N-acetyl-alpha-D-glucosamine
-
 Pasteurella multocida H+ + N-acetyl-beta-D-glucosaminyl-(1->4)-[hyaluronan](n) + UDP
-
 ?

Synonyms

Synonyms Comment Organism
HyaD
-
 Pasteurella multocida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
additional information
-
 highest activity in MES, MOPS and HEPES buffers and lower activity in TRIS-buffer Pasteurella multocida
5.5 8 recombinant His-taggged protein, residues 1-703 Pasteurella multocida