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Literature summary for 3.2.1.B16 extracted from

  • Nomura, T.; Quesada, A.L.; Kutchan, T.M.
    The new beta-D-glucosidase in terpenoid-isoquinoline alkaloid biosynthesis in Psychotria ipecacuanha (2008), J. Biol. Chem., 283, 34650-34659.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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

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 [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

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Carapichea ipecacuanha 5829
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
61800
-
calculated from sequence Carapichea ipecacuanha

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Carapichea ipecacuanha B6ZKM3
-
-

Purification (Commentary)

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

Source Tissue Comment Organism Textmining
root
-
Carapichea ipecacuanha
-
shoot
-
Carapichea ipecacuanha
-

Substrates and Products (Substrate)

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

Subunits Comment Organism
? x * 61800, calculated from sequence Carapichea ipecacuanha

Synonyms

Synonyms Comment Organism
Ipecac alkaloid beta-D-glucosidase
-
Carapichea ipecacuanha
IpeGlu1
-
Carapichea ipecacuanha

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Carapichea ipecacuanha
55 60
-
Carapichea ipecacuanha

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
substrate: 1alpha(S)-N-deacetylisoipecoside or 1beta(R)-N-deacetylipecoside Carapichea ipecacuanha

pI Value

Organism Comment pI Value Maximum pI Value
Carapichea ipecacuanha calculated from sequence
-
6.26

kcat/KM [mM/s]

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