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

BRENDA support

Literature summary for 2.4.1.4 extracted from

  • Cambon, E.; Barbe, S.; Pizzut-Serin, S.; Remaud-Simeon, M.; Andre, I.
    Essential role of amino acid position 226 in oligosaccharide elongation by amylosucrase from Neisseria polysaccharea (2014), Biotechnol. Bioeng., 111, 1719-1728.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain JM109 Neisseria polysaccharea

Protein Variants

Protein Variants Comment Organism
E328Q site-directed mutagenesis, inactive mutant Neisseria polysaccharea
R226N site-directed mutagenesis, compared to the wild-type enzyme, the mutant shows a 10fold enhancement in the catalytic efficiency and a nearly twofold higher production of an insoluble amylose-like polymer Neisseria polysaccharea
R226X site-directed mutagenesis, the single site mutants, except R226N, show reduced activity compare to the wild-type enzyme Neisseria polysaccharea

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0123
-
maltotriose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
0.014
-
maltobiose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
0.024
-
maltopentaose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
0.026
-
maltotetraose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
11
-
maltobiose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
16
-
maltotriose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
30
-
maltotetraose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
61
-
maltopentaose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Neisseria polysaccharea amylosucrase from Neisseria polysaccharea is a transglucosylase that synthesizes an insoluble amylose-like polymer from sole substrate sucrose, product isolation and analysis, overview ?
-
?
sucrose + maltobiose Neisseria polysaccharea
-
D-fructose + maltotriose
-
?
sucrose + maltopentaose Neisseria polysaccharea
-
D-fructose + maltohexaose
-
?
sucrose + maltotetraose Neisseria polysaccharea
-
D-fructose + maltopentaose
-
?
sucrose + maltotriose Neisseria polysaccharea
-
D-fructose + maltotetraose
-
?

Organism

Organism UniProt Comment Textmining
Neisseria polysaccharea
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information amylosucrase from Neisseria polysaccharea is a transglucosylase that synthesizes an insoluble amylose-like polymer from sole substrate sucrose, product isolation and analysis, overview Neisseria polysaccharea ?
-
?
sucrose + maltobiose
-
Neisseria polysaccharea D-fructose + maltotriose
-
?
sucrose + maltopentaose
-
Neisseria polysaccharea D-fructose + maltohexaose
-
?
sucrose + maltotetraose
-
Neisseria polysaccharea D-fructose + maltopentaose
-
?
sucrose + maltotriose
-
Neisseria polysaccharea D-fructose + maltotetraose
-
?

Subunits

Subunits Comment Organism
More amylosucrases adopt a deep pocket topology of about 15 A with the catalytic triad located at the bottom Neisseria polysaccharea

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Neisseria polysaccharea

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1
-
maltobiose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
3.1
-
sucrose with maltobiose, pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
4.8
-
maltotriose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
4.8
-
sucrose with maltotriose, pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
7.5
-
maltotetraose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
7.5
-
sucrose with maltotetraose, pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
14
-
maltobiose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
14
-
sucrose with maltobiose, pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
33
-
maltotriose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
33
-
sucrose with maltotriose, pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
47
-
maltopentaose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
47
-
sucrose with maltopentaose, pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
67
-
maltotetraose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
67
-
sucrose with maltotetraose, pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
173
-
maltopentaose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
173
-
sucrose with maltopentaose, pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Neisseria polysaccharea

General Information

General Information Comment Organism
evolution the enzyme belongs to the GH13 family. Amylosucrases adopt a deep pocket topology of about 15 A with the catalytic triad located at the bottom Neisseria polysaccharea
additional information active site topology and structure coparisons, overview Neisseria polysaccharea
additional information Arg226, was proposed from molecular modeling studies to play an important role in the formation of the active site topology and in the accessibility of ligands to the catalytic site Neisseria polysaccharea

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.25
-
maltotetraose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
0.28
-
maltobiose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
0.3
-
maltotriose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
0.77
-
maltopentaose pH 7.0, 30°C, recombinant wild-type enzyme Neisseria polysaccharea
0.99
-
maltobiose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
2.6
-
maltotetraose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
2.7
-
maltotriose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea
7.2
-
maltopentaose pH 7.0, 30°C, recombinant mutant R226N Neisseria polysaccharea