Cloned (Comment) | Organism |
---|---|
expression of maltose-binding protein fusion HepI, MBP-HepI, in Escherichia coli | Pedobacter heparinus |
Protein Variants | Comment | Organism |
---|---|---|
C297S | site-directed mutagenesis, the mutant suppresses the dimerization and shows 70% reduced activity compared to the wild-type enzyme | Pedobacter heparinus |
General Stability | Organism |
---|---|
after freeze-thawing, 97% of maximal activity remains | Pedobacter heparinus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
alpha-lactose | - |
Pedobacter heparinus | |
Ca2+ | - |
Pedobacter heparinus | |
Dextran | - |
Pedobacter heparinus | |
DTT | suppresses the dimerization | Pedobacter heparinus | |
glycerol | - |
Pedobacter heparinus | |
PEG 20000 | - |
Pedobacter heparinus | |
PEG 8000 | - |
Pedobacter heparinus | |
sucrose | - |
Pedobacter heparinus | |
trehalose | - |
Pedobacter heparinus | |
Triton X-100 | - |
Pedobacter heparinus | |
Tween 80 | - |
Pedobacter heparinus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic modeling based on inactivation data, overview | Pedobacter heparinus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activates | Pedobacter heparinus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pedobacter heparinus | - |
formerly Flavobacterium heparinum | - |
Purification (Comment) | Organism |
---|---|
recombinant maltose-binding protein fusion HepI, MBP-HepI, from Escherichia coli by affinity and anion exchange chromatography, followed by gel filtration to apparent homogeneity | Pedobacter heparinus |
Renatured (Comment) | Organism |
---|---|
temperature-induced reactivation of MBP-HepI, when the temperature is lowered from 35°C to 4°C, the rate constant of unfolding decreases by 6000times while that of refolding decreases by only 600times, MBP-HepI undergoes reactivation during the cooling treatment at 4°C after incubation at 35°C | Pedobacter heparinus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
149 | - |
purified recombinant HepI, pH 7.5, 30°C | Pedobacter heparinus |
Storage Stability | Organism |
---|---|
4°C, purified recombinant MBP-HEPI, 1 week, 95% remaining activity | Pedobacter heparinus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | cleavage/degradation of heparin forming unsaturated uronic acid | Pedobacter heparinus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional model structure, modelling based on the known crystal structure of Bacteroides thetaiotaomicron HepI, overview | Pedobacter heparinus |
Synonyms | Comment | Organism |
---|---|---|
heparinase I | - |
Pedobacter heparinus |
HepI | - |
Pedobacter heparinus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Pedobacter heparinus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
the inactivation shows strong concentration dependence. Acceleration of the inactivation with the increase of enzyme concentration implies multi-molecular interactions in the thermal inactivation. Strong reversibility of the unfolding of MBP-HepI, overview | Pedobacter heparinus |
30 | - |
purified recombinant enzyme, half-life is 160 min | Pedobacter heparinus |
70 | - |
purified recombinant enzyme, loss of 97% activity within 1 min | Pedobacter heparinus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pedobacter heparinus |
General Information | Comment | Organism |
---|---|---|
additional information | intermolecular disulfide bond formation in HepI important for catalysis, overview | Pedobacter heparinus |