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Literature summary for 3.2.1.182 extracted from

  • Sue, M.; Nakamura, C.; Miyamoto, T.; Yajima, S.
    Active-site architecture of benzoxazinone-glucoside beta-D-glucosidases in Triticeae (2011), Plant Sci., 180, 268-275.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as His-tagged fusion protein Triticum aestivum
expressed in Escherichia coli as His-tagged fusion protein Secale cereale

Crystallization (Commentary)

Crystallization (Comment) Organism
crystal structures of an inactive mutant of the wheat glucosidase complexed with the natural substrate DIMBOA-Glc, wheat and rye glucosidases complexed with an aglycone DIMBOA, and wheat and rye glucosidases complexed with an inhibitor 2-fluoro-2- deoxy-beta-D-glucose are analyzed Triticum aestivum
crystal structures of an inactive mutant of the wheat glucosidase complexed with the natural substrate DIMBOA-Glc, wheat and rye glucosidases complexed with an aglycone DIMBOA, and wheat and rye glucosidases complexed with an inhibitor 2-fluoro-2- deoxy-beta-D-glucose are analyzed Secale cereale

Protein Variants

Protein Variants Comment Organism
E191A crystals of the enzymatically inactive mutant E191A are soaked in a solution containing DIMBOA-Glc as substrate and 3.5M sodium formate as cryoprotectant, the structure is solved at a resolution of 2.2 A and the DIMBOA-Glc molecule is clearly defined. In the structure, the aglycone moiety is shown to be stabilized by interaction with the aromatic ring of W379 and by a hydrogen bond with T194 Triticum aestivum
E462A inactive mutant, E462 plays a crucial role in hydrolysis of the substrate Triticum aestivum
G464S/S465L mutation reduces the reaction efficiency toward DIBOA-Glc to 36% of wild type, whereas it enhances efficiency toward DIMBOA-Glc to 400% of wild type Secale cereale

Inhibitors

Inhibitors Comment Organism Structure
2-fluoro-2-deoxy-beta-D-glucose
-
Secale cereale
2-fluoro-2-deoxy-beta-D-glucose
-
Triticum aestivum

Organism

Organism UniProt Comment Textmining
Secale cereale Q9FYS3
-
-
Triticum aestivum
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using HisTrap HP column and and Superdex 200 HR Triticum aestivum
using HisTrap HP column and and Superdex 200 HR Secale cereale

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-hydroxy-3-oxo-3,4,dihydro-2H-1,4,benzoxazin-2-yl beta-D-gluco-pyranoside + H2O
-
Secale cereale 2,4-dihydroxy-2H-1,4-benzoxazin-3(4H)-one + beta-D-glucose
-
?
DIBOA-beta-D-glucoside + H2O TaGlu1b hydrolyzes DIMBOA-Glc much better than DIBOA-Glc Triticum aestivum DIBOA + beta-D-glucose
-
?
DIBOA-glucoside + H2O ScGlu shows similar enzyme activity toward DIMBOA-Glc and DIBOA-Glc Secale cereale DIBOA + beta-D-glucose
-
?
DIMBOA-beta-D-glucoside + H2O TaGlu1b hydrolyzes DIMBOA-Glc much better than DIBOA-Glc Triticum aestivum DIMBOA + beta-D-glucose
-
?
DIMBOA-glucoside + H2O ScGlu shows similar enzyme activity toward DIMBOA-Glc and DIBOA-Glc Secale cereale DIMBOA + beta-D-glucose
-
?
esculin + H2O
-
Secale cereale esculetin + beta-D-glucose
-
?
genistin + H2O
-
Secale cereale ?
-
?

Synonyms

Synonyms Comment Organism
benzoxazinone-glucoside beta-D-glucosidase
-
Triticum aestivum
benzoxazinone-glucoside beta-D-glucosidase
-
Secale cereale
ScGlu
-
Secale cereale
TaGlu1
-
Triticum aestivum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Triticum aestivum
30
-
assay at Secale cereale

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
assay at Triticum aestivum
5.5
-
assay at Secale cereale

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
120
-
DIMBOA-glucoside pH and temperature not specified in the publication Secale cereale
150
-
DIBOA-glucoside pH and temperature not specified in the publication Secale cereale
150
-
DIBOA-beta-D-glucoside pH and temperature not specified in the publication Triticum aestivum
4100
-
DIMBOA-beta-D-glucoside pH and temperature not specified in the publication Triticum aestivum