Information on EC 3.7.1.4 - phloretin hydrolase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
3.7.1.4
-
RECOMMENDED NAME
GeneOntology No.
phloretin hydrolase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
phloretin + H2O = phloretate + phloroglucinol
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ester hydrolysis
hydrolysis of C-C bond
retro-Friedel-Crafts acylation
-
-
SYSTEMATIC NAME
IUBMB Comments
2',4,4',6'-tetrahydroxydehydrochalcone 1,3,5-trihydroxybenzenehydrolase
Also hydrolyses other C-acylated phenols related to phloretin.
CAS REGISTRY NUMBER
COMMENTARY hide
37289-38-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
wK1
gene phy; strain wK1
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
129Sv background
-
-
Manually annotated by BRENDA team
strain Y46
-
-
Manually annotated by BRENDA team
strain Y46
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
physiological function
additional information
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comparison of the enzyme phloretin hydrolase Phy from Eubacterium ramulus with the diacetyl phloroglucinol hydrolase (Phlg) from Pseudomonas fluorescens, which catalyses a similar, hydrolytic, de-acylation of its substrate, homology modeling of Phy based on the structure of Phlg, PDB ID 3HWP
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2',3,4',6'-tetrahydroxy-4-methoxydihydrochalcone + H2O
phloroglucinol + ?
show the reaction diagram
-
-
-
?
2',3,4',6'-tetrahydroxy-dihydrochalcone + H2O
phloroglucinol + ?
show the reaction diagram
-
-
-
?
2',3,4,4',6'-pentahydroxy-dihydrochalcone 2'-beta-D-glucoside + H2O
phloroglucinol beta-D-glucoside + 3-(3,4-dihydroxyphenyl)propanoic acid
show the reaction diagram
-
-
-
?
2',3,4,4',6'-tetrahydroxy-dihydrochalcone + H2O
phloroglucinol + ?
show the reaction diagram
-
-
-
?
2',4'-dihydroxy-4-methoxydihydrochalcone + H2O
resorcinol + ?
show the reaction diagram
-
-
-
?
2',4'-dihydroxypropiophenone + H2O
resorcinol
show the reaction diagram
-
-
-
?
2',4,4'-trihydroxyhydrochalcone
?
show the reaction diagram
-
-
-
-
?
2',6'-dihydroxy-4,4'-dimethoxydihydrochalcone + H2O
3-methoxyresorcinol + ?
show the reaction diagram
-
-
-
?
3'-methylchloracetophenone + H2O
?
show the reaction diagram
-
-
-
-
?
baohuoside I + H2O
?
show the reaction diagram
beta-methyl D-glucoside + H2O
?
show the reaction diagram
-
-
-
-
?
cellobiose + H2O
D-glucose
show the reaction diagram
-
-
-
?
cerebrosides + H2O
?
show the reaction diagram
-
-
-
-
?
chloracetophenone + H2O
?
show the reaction diagram
-
-
-
-
?
epimedin A + H2O
?
show the reaction diagram
epimedin B + H2O
?
show the reaction diagram
epimedin C + H2O
?
show the reaction diagram
-
-
-
?
icariin + H2O
?
show the reaction diagram
lactose + H2O
?
show the reaction diagram
-
66% reduced activity towards the substrate in rats fed a low starch diet as compared to rats fed a high starch diet
-
-
?
lactose + H2O
beta-D-galactose + beta-D-glucose
show the reaction diagram
lactose + H2O
D-galactose + D-glucose
show the reaction diagram
lactose + H2O
D-glucose + D-galactose
show the reaction diagram
lactose + H2O
glucose + galactose
show the reaction diagram
-
-
-
-
?
N-palmitoyldihydroglucosylceramide + H2O
?
show the reaction diagram
-
-
-
-
?
o-nitrophenyl-beta-D-galactoside + H2O
o-nitrophenol + beta-D-galactose
show the reaction diagram
-
-
-
?
o-nitrophenyl-beta-D-glucoside + H2O
o-nitrophenol + beta-D-glucose
show the reaction diagram
-
-
-
?
p-nitrophenyl-beta-D-galactoside + H2O
p-nitrophenol + beta-D-galactose
show the reaction diagram
-
-
-
?
p-nitrophenyl-beta-D-glucoside + H2O
p-nitrophenol + beta-D-glucose
show the reaction diagram
-
-
-
?
phloretin + H2O
phloretate + phloroglucinol
show the reaction diagram
phloretin + H2O
phloroglucinol + 3-(4-hydroxyphenyl)propionic acid
show the reaction diagram
phlorhizin + H2O
3-(4-hydroxyphenyl)propanoate + arbutin
show the reaction diagram
pyridoxine-5'-beta-D-glucoside + H2O
pyridoxine + D-glucose
show the reaction diagram
pyridoxine-5'-beta-D-glucoside + lactose
pyridoxine + beta-D-glucose + pyridoxine disaccharide
show the reaction diagram
-
-
product identification, product is formed by a pyridoxine molecule with the 5'-beta-gkucoside and another glucose or galactose moiety
-
-
salicin + H2O
?
show the reaction diagram
-
-
-
-
?
sucrose + H2O
D-fructose + D-glucose
show the reaction diagram
-
87% reduced activity towards the substrate in rats fed a low starch diet as compared to rats fed a high starch diet
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
baohuoside I + H2O
?
show the reaction diagram
epimedin A + H2O
?
show the reaction diagram
epimedin B + H2O
?
show the reaction diagram
epimedin C + H2O
?
show the reaction diagram
Q02401
-
-
-
?
icariin + H2O
?
show the reaction diagram
Q02401
the metabolic pathways of icariin by rat intestinal flora and enzyme solution are basically the same, including 3-O-rhamnose, 7-O-glucose hydrolysis or dual 3, 7-hydrolysis. Compared to 3-O-rhamnose and dual hydrolysis of 3, 7-, the 7-O-glucose hydrolysis is easier. The yielded metabolites contain M1 (icariside I), M2 (icaritin), and baohuoside I
-
-
?
phloretin + H2O
phloretate + phloroglucinol
show the reaction diagram
phloretin + H2O
phloroglucinol + 3-(4-hydroxyphenyl)propionic acid
show the reaction diagram
-
Q715L4
during flavonoid degradation
-
-
?
pyridoxine-5'-beta-D-glucoside + H2O
pyridoxine + D-glucose
show the reaction diagram
-
enzyme might play an important role in providing the bioavailability of pyridoxine-5'-beta-D-glucoside
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zn2+
-
a Zn-dependent C-C hydrolase, His123, His251, Glu154 and Glu255 are conserved zinc binding residues
additional information
-
no affected by Mg2+ and Ca2+; possibly a metal-dependent enzyme, no effects by Ca2+ or Mg2+ on the enzyme activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-bromopyrrolidine-2,5-dione
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3% inhibition at 1 mM
2',4'-dinitrophenyl-3-fluoro-2-deoxy-beta-D-glucopyranoside
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specific 100fold inhibition of phloretin and lactose hydrolase activity, no inhibition of pyridoxine-5'-beta-D-glucoside hydrolase activity
3-(4-hydroxyphenyl)propionic acid
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i.e. phloretate, 30% product inhibition at 0.12 mM; product inhibition
Ag+
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0.02 mM
Al2(SO4)3
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1 mM
beta-D-glucose
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inhibition of pyridoxine-5'-beta-D-glucoside hydrolase activity
brain cerebroside
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competitive inhibition
-
cellobiose
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inhibition of pyridoxine-5'-beta-D-glucoside hydrolase activity
CuCl2
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85% inhibition at 1 mM
gluconolactone
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L-cysteine
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in presence of p-chloromercuribenzoate, 0.05 mM
lactose
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inhibition of pyridoxine-5'-beta-D-glucoside hydrolase activity, competitive
maltose
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inhibition of pyridoxine-5'-beta-D-glucoside hydrolase activity, inhibition is reduced to 60-75% by addition of 1-deoxynojirimycin
N-bromosuccinimide
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97% inhibition at 1 mM
N-ethylmaleimide
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65% inhibition at 1 mM
Na2MoO4
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1 mM
Na2WO4
-
1 mM
NEM
-
35% inhibition at 1 mM
o-phenanthroline
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20% inhibition at 1 mM
p-chloromercuribenzoate
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0.05 mM
phlorizin
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competitive inhibition
phloroglucinol
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46% product inhibition at 0.12 mM; product inhibition
thyroid hormone
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LPH mRNA level and lactase activity repressed by the injection of thyroid hormones during starvation
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Zn2+
-
0.2 mM
additional information
-
no inhibition by pyridoxine, trehalose, and sucrose
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
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-
cysteine
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dithiothreitol
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Gata4
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activates the LPD promoter, important positional signal required for the maintenance of jejunal-ileal identities in the adult mouse small intestine
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Gata6
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activates the LPD promoter
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iodoacetate
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N-ethylmaleimide
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-
p-chloromercuribenzoate
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-
Sucrose
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is capable of enhancing lactase gene expression in starved rats when they have a sustainable thyroid hormone level
additional information
-
not activated by Gatadeltaex2
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.15
3'-methylphloracetophenone
-
-
-
2.78
cellobiose
-
-
16 - 21.8
lactose
0.013 - 0.4
phloretin
0.4
phlorizin
-
-
1
pyridoxine-5'-beta-D-glucoside
-
pH 6.0, 37°C
additional information
additional information
-
kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
47
lactose
Rattus norvegicus
-
pH 6.0, 37°C
10
phloretin
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Q715L4
pH 7.0, 37°C; recombinant enzyme, pH 7.0, 37°C
1
pyridoxine-5'-beta-D-glucoside
Rattus norvegicus
-
pH 6.0, 37°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
64
beta-D-glucose
-
pH 6.0, 37°C
56
lactose
-
pH 6.0, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0064
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cell extract
0.45
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8.7
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purified enzyme; purified recombinant enzyme, pH 7.0, 37°C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
-
assay at
6.7 - 6.8
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-
7
-
-
7.5
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.9 - 9.5
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
85% reduced LPH mRNA level in rats fed a low starch diet as compared to rats fed a high starch diet
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
LPH traverses endosomal compartments before entering the apical membrane (fluorescence microscopic and biochemical approach)
Manually annotated by BRENDA team
-
the recombinant precusor in COS-1 cells resides in the ER, not mature wild-type
Manually annotated by BRENDA team
-
LPH traverses endosomal compartments before entering the apical membrane (fluorescence microscopic and biochemical approach)
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30846
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2 * 30846, sequence calculation, 2 * 30000, recombinant enzyme, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 135000-140000, SDS-PAGE
homodimer
pentamer
-
SDS-PAGE
additional information
-
the mature enzyme shows a temperature-sensitive conformation, interaction with ER chaperones, immunglobulin binding protein, and calnexin, enzyme structure, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
mannose-rich, complex glycosylated mature enzyme, dependent on ooptimal folding of the processed proform in presence of the pro-peptide
proteolytic modification
-
enzyme is sythesized as a transport-competent precursor, i.e. pro-LPH, intracellular cleavage process in the trans-Golgi network between Arg734 and Leu 735 to yield the mature enzyme, i.e. LPHbetainitial, the propeptide of the zymogen, i.e. LPHalpha, modulates as a chaperone the folding of the mature enzyme after it has been cleaved off, mechanism, overview
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
the mature enzyme shows a temperature-sensitive conformation
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
300fold from intestine brushh border membrane
-
recombinant enzyme 13.2fold from Escherichia coli strain DH5alpha(pPH3) by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography, followed by gel filtration; recombinant enzyme from Escherichia coli, about 13.2fold
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recombinant mature enzyme and pro-enzyme from COS-1 cells
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recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3)
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, expression in Escherichia coli DH5alpha; gene phy, encoded in the 2,4-diacetylphloroglucinol operon, library screening, DNA and amino acid sequence determination and analysis, sequence comparisons and promoter sequence determination, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha(pPH3)
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gene phy, DNA and amino acid sequence determination and analysis, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
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generation of a galectin-3 knock-out mutant
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the C/T-13910 SNP in the gene encoding lactase-phlorizin hydrolase on chromosome 2 defines adult-type hypolactasia status. Study in young finns analysing association of adult-type hypolactasia with coronary artery disease risk factors and with with markers of early atherosclerosis respectively
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transient expression of mature enzyme and pro-enzyme in COS-1 cells, in vitro transcritption and translation
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transient transfection of deletion constructs in both LPH-expressing Caco-2 and LPH-nonexpressing Hep-G2 cells, and in HeLa cells
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A126S
-
site-directed mutagenesis
F218Y
-
site-directed mutagenesis
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
molecular biology