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Literature summary 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 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.4.1.357 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)

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

General Stability

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.1.357 3-hydroxyphloridzin
-
 Malus domestica
2.4.1.357 Al3+ 67.81% residual activity at 5 mM Malus domestica
2.4.1.357 Ca2+ 54.21% residual activity at 5 mM Malus domestica
2.4.1.357 Co2+ complete inhibition at 5 mM Malus domestica
2.4.1.357 Cu2+ complete inhibition at 5 mM Malus domestica
2.4.1.357 Fe3+ 60.78% residual activity at 5 mM Malus domestica
2.4.1.357 Mn2+ 47.41% residual activity at 5 mM Malus domestica
2.4.1.357 additional information negligible inhibition by uridine, K+, Ni2+, and Ba2+ Malus domestica
2.4.1.357 N-ethylmaleimide 20.53% residual activity at 1 mM Malus domestica
2.4.1.357 p-hydroxymercuribenzoate 18.45% residual activity at 1 mM Malus domestica
2.4.1.357 trilobatin
-
 Malus domestica
2.4.1.357 UDP complete inhibition at 5 mM Malus domestica
2.4.1.357 UMP
-
 Malus domestica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.357 0.0005
-
 phloretin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.00106
-
 quercetin with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.00181
-
 quercetin with UDP-alpha-D-galactose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.00209
-
 kaempferol with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.0024
-
 isoliquiritigenin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.00289
-
 luteolin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.00409
-
 phloretin with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.00753
-
 UDP-alpha-D-glucose with phloretin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.01457
-
 ADP-alpha-D-glucose with quercetin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.02619
-
 UDP-alpha-D-galactose with quercetin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.03043
-
 UDP-alpha-D-glucose with kaempferol as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.03619
-
 UDP-alpha-D-glucose with isoliquiritigenin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.04259
-
 UDP-alpha-D-glucose with luteolin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 0.05867
-
 ADP-alpha-D-glucose with phloretin as cosubstrate, at pH 7.5 and 30°C Malus domestica

Metals/Ions

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.1.357 65300
-
 deglycosylated protein, gel filtration Malus domestica
2.4.1.357 70700
-
 native protein, gel filtration Malus domestica

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.357 Malus domestica D3UAG5 cultivar Golden Delicious
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
2.4.1.357 glycoprotein
-
 Malus domestica

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.357 30
-
-
 Malus domestica

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.1.357 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.357 2.95
-
 phloretin with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 3.24
-
 luteolin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 4.35
-
 isoliquiritigenin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 5.65
-
 kaempferol with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 6.18
-
 quercetin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 7.41
-
 quercetin with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 9.87
-
 phloretin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 14.57
-
 ADP-alpha-D-glucose with quercetin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 21.33
-
 UDP-alpha-D-glucose with phloretin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 26.19
-
 UDP-alpha-D-galactose with quercetin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 30.43
-
 UDP-alpha-D-glucose with kaempferol as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 36.19
-
 UDP-alpha-D-glucose with isoliquiritigenin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 42.59
-
 UDP-alpha-D-glucose with luteolin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 58.67
-
 ADP-alpha-D-glucose with phloretin as cosubstrate, at pH 7.5 and 30°C Malus domestica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.357 8
-
-
 Malus domestica

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.4.1.357 7 9 up to 80% of its maximal activity remains within the pH range of 7.0-9.0 Malus domestica

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.4.1.357 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

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.4.1.357 0.01
-
 at pH 7.5 and 30°C Malus domestica Cu2+
2.4.1.357 0.05
-
 at pH 7.5 and 30°C Malus domestica Co2+
2.4.1.357 0.1
-
 at pH 7.5 and 30°C Malus domestica UDP
2.4.1.357 0.24
-
 at pH 7.5 and 30°C Malus domestica p-hydroxymercuribenzoate
2.4.1.357 0.28
-
 at pH 7.5 and 30°C Malus domestica N-ethylmaleimide
2.4.1.357 2.5
-
 at pH 7.5 and 30°C Malus domestica 3-hydroxyphloridzin
2.4.1.357 3
-
 at pH 7.5 and 30°C Malus domestica trilobatin
2.4.1.357 4
-
 at pH 7.5 and 30°C Malus domestica UMP
2.4.1.357 4.5
-
 at pH 7.5 and 30°C Malus domestica Mn2+
2.4.1.357 5.5
-
 at pH 7.5 and 30°C Malus domestica Ca2+
2.4.1.357 6.5
-
 at pH 7.5 and 30°C Malus domestica Fe3+
2.4.1.357 8
-
 at pH 7.5 and 30°C Malus domestica Al3+

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.4.1.357 80
-
 ADP-alpha-D-glucose with phloretin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 140
-
 UDP-alpha-D-glucose with luteolin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 240
-
 UDP-alpha-D-glucose with isoliquiritigenin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 360
-
 UDP-alpha-D-glucose with kaempferol as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 500
-
 UDP-alpha-D-galactose with quercetin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 720
-
 phloretin with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 1040
-
 ADP-alpha-D-glucose with quercetin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 1120
-
 luteolin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 1810
-
 isoliquiritigenin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 2700
-
 kaempferol with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 2830
-
 UDP-alpha-D-glucose with phloretin as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 5240
-
 quercetin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 6990
-
 quercetin with ADP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica
2.4.1.357 19730
-
 phloretin with UDP-alpha-D-glucose as cosubstrate, at pH 7.5 and 30°C Malus domestica