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Literature summary for 2.4.1.10 extracted from

  • Batista, F.R.; Hernandez, L.; Fernandez, J.R.; Arrieta, J.; Menendez, C.; Gomez, R.; Tambara, Y.; Pons, T.
    Substitution of Asp-309 by Asn in the Arg-Asp-Pro (RDP) motif of Acetobacter diazotrophicus levansucrase affects sucrose hydrolysis, but not enzyme specificity (1999), Biochem. J., 337, 503-506.
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Cloned(Commentary)

Cloned (Comment) Organism
cloning and expression of LsdA in a levansucrase deficient Acetobacter diazotrophicus mutant strain Gluconacetobacter diazotrophicus

Protein Variants

Protein Variants Comment Organism
D309N site-directed mutagenesis of RDP-motif, no affection of enzyme secretion but hydrolysis activity, 13fold increase of kcat, 71fold decrease of kcat/Km, unaltered Km for sucrose Gluconacetobacter diazotrophicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
10.5
-
 sucrose mutant D309N Gluconacetobacter diazotrophicus
11.4
-
 sucrose wild-type enzyme Gluconacetobacter diazotrophicus

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
 Gluconacetobacter diazotrophicus
-
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
additional information
-
 alignment of RDP-motif sequence Gluconacetobacter diazotrophicus

Organism

Organism UniProt Comment Textmining
Gluconacetobacter diazotrophicus Q43998 i.e. Acetobacter diazotrophicus
-
Gluconacetobacter diazotrophicus Q43998 constitutive LsdA, strain SRT4
-

Purification (Commentary)

Purification (Comment) Organism
recombinant and native wild-type and recombinant mutant from supernatant Gluconacetobacter diazotrophicus

Reaction

Reaction Comment Organism Reaction ID
sucrose + [6)-beta-D-fructofuranosyl-(2->]n alpha-D-glucopyranoside = D-glucose + [6)-beta-D-fructofuranosyl-(2->]n+1 alpha-D-glucopyranoside Asp309 is involved in catalytic ping-pong mechanism, localization in conserved RDP-motif Gluconacetobacter diazotrophicus
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
sucrose + (2,6-beta-D-fructosyl)n beta-D-fructofuranosidase activity Gluconacetobacter diazotrophicus glucose + (2,6-beta-D-fructosyl)n+1
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1
-
 sucrose wild-type enzyme Gluconacetobacter diazotrophicus
13.3
-
 sucrose mutant D309N Gluconacetobacter diazotrophicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
 wild-type and mutant D309N Gluconacetobacter diazotrophicus