Cloned (Comment) | Organism |
---|---|
recombinant IpeGlu1 is efficiently expressed in a soluble protein fraction in Eschericha coli PlusS strain, more than 50% of the expressed protein is precipitated as an inclusion body | Carapichea ipecacuanha |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ag+ | 1 mM, 23% inhibition | Carapichea ipecacuanha | |
Co2+ | 1 mM, 14% inhibition | Carapichea ipecacuanha | |
Cu2+ | 1 mM, 50% inhibition | Carapichea ipecacuanha | |
Fe2+ | 1 mM, 54% inhibition | Carapichea ipecacuanha | |
Mg2+ | 1 mM, 20% inhibition | Carapichea ipecacuanha | |
Mn2+ | 1 mM, 14% inhibition | Carapichea ipecacuanha | |
additional information | EDTA and 2-mercaptoethanol do not affect the activity | Carapichea ipecacuanha | |
Ni2+ | 1 mM, 19% inhibition | Carapichea ipecacuanha | |
Zn2+ | 1 mM, 36% inhibition | Carapichea ipecacuanha |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
ipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
0.29 | - |
vincoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
0.32 | - |
1beta(R)-N-deacetylipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
0.39 | - |
1beta(R)-6-O-methyl-N-deacetylipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
1.3 | - |
1alpha(S)-6-O-methyl-N-deacetylisoipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
5.8 | - |
1alpha(S)-N-deacetylisoipecoside | pH 5.0, 30°C | Carapichea ipecacuanha |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Carapichea ipecacuanha | 5829 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
61800 | - |
calculated from sequence | Carapichea ipecacuanha |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
1beta(R)-6-O-methyl-N-deacetylipecoside + H2O | Carapichea ipecacuanha | the enzyme is involved in alkaloid biosynthesis | 1beta(R)-6-O-methyl-N-deacetylipecoside aglycone + D-glucose | - |
? | |
1beta(R)-N-deacetylipecoside + H2O | Carapichea ipecacuanha | the 1beta(R)-epimer N-deacetylipecoside is the precursor mainly of ipecoside and alangiside, the alkaloidal glucosides having a beta-configuration | 1beta(R)-N-deacetylipecoside aglycone + D-glucose | - |
? | |
ipecoside + H2O | Carapichea ipecacuanha | the enzyme is involved in alkaloid biosynthesis | ipecoside aglycone + D-glucose | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Carapichea ipecacuanha | B6ZKM3 | - |
- |
Purification (Comment) | Organism |
---|---|
N-terminal His-tagged IpeGlu1 enzyme is purified to homogeneity from the soluble protein fraction by metal chelation chromatography | Carapichea ipecacuanha |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
root | - |
Carapichea ipecacuanha | - |
shoot | - |
Carapichea ipecacuanha | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1alpha(S)-6-O-methyl-N-deacetylisoipecoside + H2O | kcat/KM is 16.3% of the value for 1beta(R)-N-deacetylipecoside | Carapichea ipecacuanha | 1alpha(S)-6-O-methyl-N-deacetylisoipecoside aglycone + D-glucose | - |
? | |
1alpha(S)-N-deacetylisoipecoside + H2O | kcat/KM is 16.9% of the value for 1beta(R)-N-deacetylipecoside | Carapichea ipecacuanha | 1alpha(S)-N-deacetylisoipecoside aglycone + D-glucose | - |
? | |
1beta(R)-6-O-methyl-N-deacetylipecoside + H2O | the enzyme is involved in alkaloid biosynthesis | Carapichea ipecacuanha | 1beta(R)-6-O-methyl-N-deacetylipecoside aglycone + D-glucose | - |
? | |
1beta(R)-6-O-methyl-N-deacetylipecoside + H2O | kcat/KM is 90.5% of the value for 1beta(R)-N-deacetylipecoside | Carapichea ipecacuanha | 1beta(R)-6-O-methyl-N-deacetylipecoside aglycone + D-glucose | - |
? | |
1beta(R)-N-deacetylipecoside + H2O | the 1beta(R)-epimer N-deacetylipecoside is the precursor mainly of ipecoside and alangiside, the alkaloidal glucosides having a beta-configuration | Carapichea ipecacuanha | 1beta(R)-N-deacetylipecoside aglycone + D-glucose | - |
? | |
1beta(R)-N-deacetylipecoside + H2O | IpeGlu1 preferentially hydrolyzes glucosidic Ipecac alkaloids except for their lactams, but shows poor or no activity toward other substrates, including terpenoid-indole alkaloid glucosides. IpeGlu1 activity is extremely poor toward 7-O-methyl and 6,7-O,O-dimethyl derivatives. However, 6-O-methyl derivatives are hydrolyzed as efficiently as non-methylated substrates. IpeGlu1 accepts both 1alpha(S)-N-deacetylisoipecoside and 1beta(R)-N-deacetylipecoside epimers as substrate with preference of 1beta(R)-epimer to 1alpha(S)-epimer | Carapichea ipecacuanha | 1beta(R)-N-deacetylipecoside aglycone + D-glucose | spontaneous transitions of the highly reactive aglycon. A possible mechanism is as follows: the hemiacetal aglycon forms a dialdehyde by ring opening, and the N-atom attacks the carbolyl group of the aldehyde, followed by dehydration to form an iminium cation which is considered to tautomerize to the enamine form | ? | |
ipecoside + H2O | the enzyme is involved in alkaloid biosynthesis | Carapichea ipecacuanha | ipecoside aglycone + D-glucose | - |
? | |
ipecoside + H2O | kcat/KM is 5.2fold higher than the value for 1beta(R)-N-deacetylipecoside | Carapichea ipecacuanha | ipecoside aglycone + D-glucose | - |
? | |
additional information | no activity with raucaffricine. IpeGlu1 does not accept the lactams, demethylisoalangiside and demethylalangiside, as substrate | Carapichea ipecacuanha | ? | - |
? | |
vincoside + H2O | kcat/KM is 11.1% of the value for 1beta(R)-N-deacetylipecoside | Carapichea ipecacuanha | vincoside aglycone + D-glucose | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 61800, calculated from sequence | Carapichea ipecacuanha |
Synonyms | Comment | Organism |
---|---|---|
Ipecac alkaloid beta-D-glucosidase | - |
Carapichea ipecacuanha |
IpeGlu1 | - |
Carapichea ipecacuanha |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Carapichea ipecacuanha |
55 | 60 | - |
Carapichea ipecacuanha |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
6.4 | - |
vincoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
18.7 | - |
1alpha(S)-6-O-methyl-N-deacetylisoipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
28.2 | - |
1beta(R)-N-deacetylipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
31.1 | - |
1beta(R)-6-O-methyl-N-deacetylipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
51.6 | - |
ipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
86.3 | - |
1alpha(S)-N-deacetylisoipecoside | pH 5.0, 30°C | Carapichea ipecacuanha |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
substrate: 1alpha(S)-N-deacetylisoipecoside or 1beta(R)-N-deacetylipecoside | Carapichea ipecacuanha |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Carapichea ipecacuanha | calculated from sequence | - |
6.26 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
14.4 | - |
1alpha(S)-6-O-methyl-N-deacetylisoipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
14.9 | - |
1alpha(S)-N-deacetylisoipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
22.1 | - |
vincoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
79.7 | - |
1beta(R)-6-O-methyl-N-deacetylipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
88.1 | - |
1beta(R)-N-deacetylipecoside | pH 5.0, 30°C | Carapichea ipecacuanha | |
516 | - |
ipecoside | pH 5.0, 30°C | Carapichea ipecacuanha |