Literature summary for 2.4.1.357 extracted from

Zhang, T.; Liang, J.; Wang, P.; Xu, Y.; Wang, Y.; Wei, X.; Fan, M.
Purification and characterization of a novel phloretin-2'-O-glycosyltransferase favoring phloridzin biosynthesis (2016), Sci. Rep., 6, 35274 .

Search for more literature for 2.4.1.357

Activating Compound

Activating Compound	Comment	Organism	Structure
additional information	not influenced by 20% (v/v) methanol, 20% (v/v) dimethyl sulfoxide, 5% (v/v) glycerol, 0.5 mM 4-(2-aminoethyl) benzenesulfonyl fluoride, 5 mM beta-mercaptoethanol, 5 mM dithiohtreitol, 0.5 mM NaCl, and 5 mM EDTA	Malus domestica

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Pichia pastoris strain GS115	Malus domestica

General Stability

General Stability	Organism
purified enzyme exhibits higher activity and stability in Tris/HCl buffer than in potassium phosphate buffer at pH 7.0	Malus domestica

Inhibitors

Inhibitors	Comment	Organism	Structure
3-hydroxyphloridzin	-	Malus domestica
Al3+	67.81% residual activity at 5 mM	Malus domestica
Ca2+	54.21% residual activity at 5 mM	Malus domestica
Co2+	complete inhibition at 5 mM	Malus domestica
Cu2+	complete inhibition at 5 mM	Malus domestica
Fe3+	60.78% residual activity at 5 mM	Malus domestica
Mn2+	47.41% residual activity at 5 mM	Malus domestica
additional information	negligible inhibition by uridine, K+, Ni2+, and Ba2+	Malus domestica
N-ethylmaleimide	20.53% residual activity at 1 mM	Malus domestica
p-hydroxymercuribenzoate	18.45% residual activity at 1 mM	Malus domestica
trilobatin	-	Malus domestica
UDP	complete inhibition at 5 mM	Malus domestica
UMP	-	Malus domestica

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0005	-	phloretin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.00106	-	quercetin	with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.00181	-	quercetin	with UDP-alpha-D-galactose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.00209	-	kaempferol	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.0024	-	isoliquiritigenin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.00289	-	luteolin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.00409	-	phloretin	with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.00753	-	UDP-alpha-D-glucose	with phloretin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.01457	-	ADP-alpha-D-glucose	with quercetin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.02619	-	UDP-alpha-D-galactose	with quercetin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.03043	-	UDP-alpha-D-glucose	with kaempferol as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.03619	-	UDP-alpha-D-glucose	with isoliquiritigenin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.04259	-	UDP-alpha-D-glucose	with luteolin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
0.05867	-	ADP-alpha-D-glucose	with phloretin as cosubstrate, at pH 7.5 and 30°C	Malus domestica

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	119.43% activity at 5 mM	Malus domestica
Mg2+	130.35% activity at 2 mM	Malus domestica

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
65300	-	deglycosylated protein, gel filtration	Malus domestica
70700	-	native protein, gel filtration	Malus domestica

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
UDP-alpha-D-glucose + phloretin	Malus domestica	-	UDP + phlorizin	-	?

Organism

Organism	UniProt	Comment	Textmining
Malus domestica	D3UAG5	cultivar Golden Delicious	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	-	Malus domestica

Purification (Commentary)

Purification (Comment)	Organism
DEAE anion exchange chromatography, His-Trap affinity chromatography, and Superdex G75 gel filtration	Malus domestica

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ADP-alpha-D-glucose + phloretin	-	Malus domestica	ADP + phlorizin	-	?
ADP-alpha-D-glucose + quercetin	-	Malus domestica	ADP + isoquercitrin	-	?
UDP-alpha-D-galactose + quercetin	-	Malus domestica	UDP + hyperoside	-	?
UDP-alpha-D-glucose + isoliquiritigenin	-	Malus domestica	UDP + isoliquiritin	-	?
UDP-alpha-D-glucose + kaempferol	-	Malus domestica	UDP + astragalin	-	?
UDP-alpha-D-glucose + luteolin	-	Malus domestica	UDP + cynaroside	-	?
UDP-alpha-D-glucose + phloretin	-	Malus domestica	UDP + phlorizin	-	?
UDP-alpha-D-glucose + quercetin	-	Malus domestica	UDP + quercetin-3-O-glucoside	-	?

Subunits

Subunits	Comment	Organism
homodimer	1 * 35200 + 1 * 24900, deglycosylated protein, SDS-PAGE	Malus domestica

Synonyms

Synonyms	Comment	Organism
P2'GT	-	Malus domestica
phloretin-2'-O-glycosyltransferase	-	Malus domestica

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	-	Malus domestica

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
25	35	the enzyme remains stable at 25-35°C for 1 h, retains 75% and 45% of its activity after 1 h at 45°C and 55°C, respectively, and is inactivated after 1 h at 65°C	Malus domestica

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
2.95	-	phloretin	with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
3.24	-	luteolin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
4.35	-	isoliquiritigenin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
5.65	-	kaempferol	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
6.18	-	quercetin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
7.41	-	quercetin	with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
9.87	-	phloretin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
14.57	-	ADP-alpha-D-glucose	with quercetin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
21.33	-	UDP-alpha-D-glucose	with phloretin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
26.19	-	UDP-alpha-D-galactose	with quercetin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
30.43	-	UDP-alpha-D-glucose	with kaempferol as cosubstrate, at pH 7.5 and 30°C	Malus domestica
36.19	-	UDP-alpha-D-glucose	with isoliquiritigenin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
42.59	-	UDP-alpha-D-glucose	with luteolin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
58.67	-	ADP-alpha-D-glucose	with phloretin as cosubstrate, at pH 7.5 and 30°C	Malus domestica

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	-	Malus domestica

pH Range

pH Minimum	pH Maximum	Comment	Organism
7	9	up to 80% of its maximal activity remains within the pH range of 7.0-9.0	Malus domestica

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
7	9	the enzyme remains stable in the pH range of 7.0-9.0 incubated at 30°C for 24 h	Malus domestica

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor	Structure
0.01	-	at pH 7.5 and 30°C	Malus domestica	Cu2+
0.05	-	at pH 7.5 and 30°C	Malus domestica	Co2+
0.1	-	at pH 7.5 and 30°C	Malus domestica	UDP
0.24	-	at pH 7.5 and 30°C	Malus domestica	p-hydroxymercuribenzoate
0.28	-	at pH 7.5 and 30°C	Malus domestica	N-ethylmaleimide
2.5	-	at pH 7.5 and 30°C	Malus domestica	3-hydroxyphloridzin
3	-	at pH 7.5 and 30°C	Malus domestica	trilobatin
4	-	at pH 7.5 and 30°C	Malus domestica	UMP
4.5	-	at pH 7.5 and 30°C	Malus domestica	Mn2+
5.5	-	at pH 7.5 and 30°C	Malus domestica	Ca2+
6.5	-	at pH 7.5 and 30°C	Malus domestica	Fe3+
8	-	at pH 7.5 and 30°C	Malus domestica	Al3+

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
80	-	ADP-alpha-D-glucose	with phloretin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
140	-	UDP-alpha-D-glucose	with luteolin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
240	-	UDP-alpha-D-glucose	with isoliquiritigenin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
360	-	UDP-alpha-D-glucose	with kaempferol as cosubstrate, at pH 7.5 and 30°C	Malus domestica
500	-	UDP-alpha-D-galactose	with quercetin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
720	-	phloretin	with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
1040	-	ADP-alpha-D-glucose	with quercetin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
1120	-	luteolin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
1810	-	isoliquiritigenin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
2700	-	kaempferol	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
2830	-	UDP-alpha-D-glucose	with phloretin as cosubstrate, at pH 7.5 and 30°C	Malus domestica
5240	-	quercetin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
6990	-	quercetin	with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica
19730	-	phloretin	with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C	Malus domestica

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Release 2023.1 (January 2023)