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

BRENDA support

Literature summary extracted from

  • Pojasek, K.; Shriver, Z.; Hu, Y.; Sasisekharan, R.
    Histidine 295 and histidine 510 are crucial for the enzymatic degradation of heparan sulfate by heparinase III (2000), Biochemistry, 39, 4012-4019.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.2.8 expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli, recombinant wild-type enzyme shows a lower activity than the native one Pedobacter heparinus

Protein Variants

EC Number Protein Variants Comment Organism
4.2.2.8 H105A PCR overlap extension site-directed mutagenesis, very low expression level, no measurement of activity possible, reduced expression level Pedobacter heparinus
4.2.2.8 H110A PCR overlap extension site-directed mutagenesis, reduced kcat, highly reduced Km compared to both recombinant and native wild-type enzymes, reduced expression level Pedobacter heparinus
4.2.2.8 H139A PCR overlap extension site-directed mutagenesis, reduced kcat and increased Km compared to both recombinant and native wild-type enzymes, reduced expression level Pedobacter heparinus
4.2.2.8 H152A PCR overlap extension site-directed mutagenesis, reduced Km and a kcat value between the recombinant and the native wild-type enzyme Pedobacter heparinus
4.2.2.8 H225A PCR overlap extension site-directed mutagenesis, Km is the same as for the recombinant wild-type, reduced kcat Pedobacter heparinus
4.2.2.8 H234A PCR overlap extension site-directed mutagenesis, Km is similar to the recombinant wild-type, reduced kcat Pedobacter heparinus
4.2.2.8 H241A PCR overlap extension site-directed mutagenesis, highly reduced kcat, highly reduced Km compared to both recombinant and native wild-type enzymes Pedobacter heparinus
4.2.2.8 H295A PCR overlap extension site-directed mutagenesis, inactive mutant Pedobacter heparinus
4.2.2.8 H36A PCR overlap extension site-directed mutagenesis, reduced Km and a kcat value between the recombinant and the native wild-type enzyme Pedobacter heparinus
4.2.2.8 H424A PCR overlap extension site-directed mutagenesis, reduced Km and kcat Pedobacter heparinus
4.2.2.8 H469A PCR overlap extension site-directed mutagenesis, reduced Km and increased kcat compared to both recombinant and native wild-type enzymes Pedobacter heparinus
4.2.2.8 H510A PCR overlap extension site-directed mutagenesis, inactive mutant Pedobacter heparinus
4.2.2.8 H539A PCR overlap extension site-directed mutagenesis, Km between the recombinant and the native wild-type enzyme, kcat is increased compared to both wild-type enzymes Pedobacter heparinus

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.2.2.8 diethyl dicarbonate inactivation, 80% reversible by hydroxylamine within 6 h, mapping of modified histidine residues Pedobacter heparinus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.2.8 0.009
-
heparan sulfate mutant H110A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 0.016
-
heparan sulfate mutant H241A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 0.058
-
heparan sulfate mutant H152A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 0.059
-
heparan sulfate mutant H424A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 0.071
-
heparan sulfate mutant H469A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 0.075
-
heparan sulfate mutant H234A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 0.08
-
heparan sulfate recombinant wild-type enzyme and mutant H225A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 0.092
-
heparan sulfate mutant H539A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 0.098
-
heparan sulfate mutant H36A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 0.143
-
heparan sulfate native wild-type enzyme, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 0.191
-
heparan sulfate mutant H139A, pH 7.6, 35°C Pedobacter heparinus

Organism

EC Number Organism UniProt Comment Textmining
4.2.2.8 Pedobacter heparinus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.2.8 recombinant His-tagged wild-type and mutants from Escherichia coli Pedobacter heparinus

Reaction

EC Number Reaction Comment Organism Reaction ID
4.2.2.8 [GlcAbeta(1-4)GlcNbeta(1-4)]n = [GlcAbeta(1-4)GlcNbeta(1-4)]n-1 + 2-amino-2-deoxy-4-O-alpha-L-threo-hex-4-enopyranuronosyl-beta-D-glucopyranose His295 and His510 are essential for catalytic activity Pedobacter heparinus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.2.8 heparan sulfate
-
Pedobacter heparinus ?
-
?

Synonyms

EC Number Synonyms Comment Organism
4.2.2.8 heparinase III
-
Pedobacter heparinus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.2.8 35
-
assay at Pedobacter heparinus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.2.8 5
-
heparan sulfate mutant H241A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 22
-
heparan sulfate mutant H225A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 23
-
heparan sulfate mutant H234A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 24
-
heparan sulfate mutant H424A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 37
-
heparan sulfate mutant H110A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 68
-
heparan sulfate mutant H139A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 78
-
heparan sulfate recombinant wild-type enzyme, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 83
-
heparan sulfate mutant H152A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 86
-
heparan sulfate mutant H36A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 94
-
heparan sulfate native wild-type enzyme, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 100
-
heparan sulfate mutant H469A, pH 7.6, 35°C Pedobacter heparinus
4.2.2.8 132
-
heparan sulfate mutant H539A, pH 7.6, 35°C Pedobacter heparinus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.2.8 7.6
-
assay at Pedobacter heparinus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
4.2.2.8 additional information
-
additional information inactivation kinetics Pedobacter heparinus