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Literature summary extracted from

  • Yanase, H.; Maeda, M.; Hagiwara, E.; Yagi, H.; Taniguchi, K.; Okamoto, K.
    Identification of functionally important amino acid residues in Zymomonas mobilis levansucrase (2002), J. Biochem., 132, 565-572.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.10 expression of wild-type and mutants in Escherichia coli Zymomonas mobilis

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.10 D194N site-directed mutagenesis, 3400fold decreased kcat and 3fold increased Km for sucrose, activity with sucrose similar to wild-type, reduced activity with raffinose and stachyose Zymomonas mobilis
2.4.1.10 D275N site-directed mutagenesis, enhanced kcat and Km for sucrose, activity with sucrose similar to wild-type, reduced activity with raffinose and stachyose Zymomonas mobilis
2.4.1.10 D302N site-directed mutagenesis, enhanced kcat and Km for sucrose, activity with sucrose similar to wild-type, reduced activity with raffinose and stachyose Zymomonas mobilis
2.4.1.10 D308N site-directed mutagenesis, enhanced kcat and Km for sucrose, activity with sucrose similar to wild-type, reduced activity with raffinose and stachyose Zymomonas mobilis
2.4.1.10 E117Q site-directed mutagenesis, slightly reduced kcat and enhanced Km for sucrose, slightly enhanced activity with sucrose, reduced activity with raffinose and stachyose Zymomonas mobilis
2.4.1.10 E211Q site-directed mutagenesis, highly reduced kcat and enhanced Km for sucrose, highly reduced activity with sucrose, reduced activity with raffinose, increased activity with stachyose Zymomonas mobilis
2.4.1.10 E278D random mutagenesis, 30fold reduced kcat and reduced Km for sucrose, slightly enhanced activity with sucrose, reduced activity with raffinose Zymomonas mobilis
2.4.1.10 E278H site-directed mutagenesis, 210fold reduced kcat and enhanced Km for sucrose, reduced activity with sucrose and reduced activity with raffinose Zymomonas mobilis
2.4.1.10 H296K site-directed mutagenesis, reduced kcat and highly increased Km for sucrose, unaltered activity with sucrose, increased activity with raffinose, highly increased activity with stachyose Zymomonas mobilis
2.4.1.10 H296L site-directed mutagenesis, reduced kcat and highly increased Km for sucrose, unaltered activity with sucrose, increased activity with raffinose, highly increased activity with stachyose Zymomonas mobilis
2.4.1.10 H296R site-directed mutagenesis, reduced kcat and highly increased Km for sucrose, unaltered activity with sucrose, increased activity with raffinose, highly increased activity with stachyose Zymomonas mobilis
2.4.1.10 H296S site-directed mutagenesis, reduced kcat and highly increased Km for sucrose, unaltered activity with sucrose, increased activity with raffinose, highly increased activity with stachyose Zymomonas mobilis
2.4.1.10 Q339H/P340A random mutagenesis, reduced activity with stachyose, slightly enhanced kcat and enhanced Km for sucrose, slightly reduced activity with sucrose Zymomonas mobilis
2.4.1.10 V223A random mutagenesis, reduced kcat, highly enhanced Km, and reduced activity with sucrose, enhanced activity with raffinose and 3.8fold with stachyose Zymomonas mobilis
2.4.1.10 W80R random mutagenesis, reduced kcat for sucrose, enhanced Km, reduced activity with sucrose, slightly reduced activity with raffinose Zymomonas mobilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.10 additional information
-
additional information mutants, overview Zymomonas mobilis
2.4.1.10 125
-
sucrose wild-type enzyme Zymomonas mobilis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.1.10 additional information
-
alignment of conserved motifs in amino acid sequence Zymomonas mobilis

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.10 Zymomonas mobilis
-
gene sucZE2, strain IFO13756
-

Reaction

EC Number Reaction Comment Organism Reaction ID
2.4.1.10 sucrose + [6)-beta-D-fructofuranosyl-(2->]n alpha-D-glucopyranoside = D-glucose + [6)-beta-D-fructofuranosyl-(2->]n+1 alpha-D-glucopyranoside Glu278, located in DXXER conserved motif, and Asp194, located in conserved RDP motif, and His296 are involved in catalysis Zymomonas mobilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.10 raffinose + (2,6-beta-D-fructosyl)n
-
Zymomonas mobilis galactose + (2,6-beta-D-fructosyl)n+1
-
?
2.4.1.10 stachyose + (2,6-beta-D-fructosyl)n
-
Zymomonas mobilis ?
-
?
2.4.1.10 sucrose + (2,6-beta-D-fructosyl)n
-
Zymomonas mobilis glucose + (2,6-beta-D-fructosyl)n+1
-
?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.10 additional information
-
additional information mutants, overview Zymomonas mobilis
2.4.1.10 28.3
-
sucrose wild-type enzyme Zymomonas mobilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.10 5
-
assay at Zymomonas mobilis