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Literature summary for 2.4.1.10 extracted from

  • Chambert, R.; Petit-Glatron, M.F.
    Polymerase and hydrolase activities of Bacillus subtilis levansucrase can be separately modulated by site-directed mutagenesis (1991), Biochem. J., 279, 35-41.
No PubMed abstract available
Search for more literature for 2.4.1.10

Cloned(Commentary)

Cloned (Comment) Organism
gene sacB from mutant strain QB252, DNA sequencing reveals an amino acid exchange R331H, expression of mutants in Escherichia coli strain XL-1 B Bacillus subtilis

Protein Variants

Protein Variants Comment Organism
H331R invers directed mutation of natural point mutation R331H to H331R restores the wild-type enzyme properties Bacillus subtilis
R331H natural point mutation, low polymerase activity, invers mutation H331R restores the wild-type enzyme properties Bacillus subtilis
R331K site-directed mutagenesis, loss of ability to perform the whole production of levan from sucrose, only capable to perform the first reaction step, the formation of the trisaccharide kestose, higher kcat than the wild-type for sucrose hydrolysis Bacillus subtilis
R331L site-directed mutagenesis, loss of ability to perform the whole production of levan from sucrose, only capable to perform the first reaction step, the formation of the trisaccharide kestose, reduced fructosyl-enzyme intermediate formation Bacillus subtilis
R331S site-directed mutagenesis, loss of ability to perform the whole production of levan from sucrose, only capable to perform the first reaction step, the formation of the trisaccharide kestose, reduced fructosyl-enzyme intermediate formation Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
40
-
 sucrose sucrose hydrolysis, wild-type Bacillus subtilis
110
-
 sucrose sucrose hydrolysis, mutant R331K Bacillus subtilis
120
-
 sucrose sucrose hydrolysis, mutant R331H Bacillus subtilis
180
-
 sucrose sucrose hydrolysis, mutants R331L and R331S Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
 sacB mutant strain QB252
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
sucrose + (2,6-beta-D-fructosyl)n polymerase and hydrolase activity can be separately modulated by site-directed mutagenesis Bacillus subtilis glucose + (2,6-beta-D-fructosyl)n+1
-
 r
sucrose + H2O hydrolysis Bacillus subtilis D-glucose + D-fructose
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0267
-
 sucrose sucrose hydrolysis, mutant R331S Bacillus subtilis
0.117
-
 sucrose sucrose hydrolysis, mutant R331L Bacillus subtilis
0.367
-
 sucrose sucrose hydrolysis, mutant R331H Bacillus subtilis
0.583
-
 sucrose sucrose hydrolysis, wild-type Bacillus subtilis
1.47
-
 sucrose sucrose hydrolysis, mutant R331K Bacillus subtilis