3.2.1.182: 4-hydroxy-7-methoxy-3-oxo-3,4-dihydro-2H-1,4-benzoxazin-2-yl glucoside beta-D-glucosidase
This is an abbreviated version!
For detailed information about 4-hydroxy-7-methoxy-3-oxo-3,4-dihydro-2H-1,4-benzoxazin-2-yl glucoside beta-D-glucosidase, go to the full flat file.
Word Map on EC 3.2.1.182
Reaction
Synonyms
benzoxazinone-glucoside beta-D-glucosidase, beta-glu, beta-glucosidase, Co-Glu, DIMBOA glucosidase, DIMBOAGlc hydrolase, GDIBOA-glucosidase, LgGLU1, LgGLU2, LgGLU3, LgGLU4, More, SbDhr1, ScGlu, TaGlu1, TaGlu1a, TaGlu1b, TaGlu1c, ZmGlu1
ECTree
3.2.1.182 4-hydroxy-7-methoxy-3-oxo-3,4-dihydro-2H-1,4-benzoxazin-2-yl glucoside beta-D-glucosidaseAdvanced search results
results (
results (Engineering
Engineering on EC 3.2.1.182 - 4-hydroxy-7-methoxy-3-oxo-3,4-dihydro-2H-1,4-benzoxazin-2-yl glucoside beta-D-glucosidase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
F198A
-
kcat/Km values of ScGlu-F198A for DIMBOA-glucoside and DIBOA-glucoside decrease by a factor of 24- to 28fold as compared to wild-type ScGlu due to an increased Km, whereas the kcat value increases
F471Y
-
kcat values for DIBOA-glucoside increased, Km (DIBOA-glucoside) increased, Km (DIMBOA-glucoside) decreased, kcat values for DIMBOA-glucoside increased
G464F
-
kcat values for DIMBOA-glucoside and DIBOA-glucoside decreased, Km values increased
G464S
-
kcat values for DIMBOA-glucoside and DIBOA-glucoside decreased, Km values increased
G464S/S465L
S465L
-
kcat values for DIBOA-glucoside decreased, Km (DIBOA-glucoside) increased, Km (DIMBOA-glucoside) decreased, kcat values for DIMBOA-glucoside increased
Y378A
-
kcat values for DIMBOA-glucoside and DIBOA-glucoside increased, Km values increased
Y378F
-
kcat values for DIMBOA-glucoside and DIBOA-glucoside increased, Km values dereased
E191A
E462A
-
inactive mutant, E462 plays a crucial role in hydrolysis of the substrate
F198A
Km increased compared to wild-type, kcat decreased compared to wild-type
F471Y
Km increased compared to wild-type, kcat (DIBOA-glucoside) increased compared to wild-type, kcat (DIMBOA-glucoside or 4-nitrophenyl beta-D-glucoside) increased compared to wild-type
S464F
Km increased compared to wild-type, kcat (DIBOA-glucoside) increased compared to wild-type, kcat (DIMBOA-glucoside or 4-nitrophenyl beta-D-glucoside) increased compared to wild-type
Y378A
Km increased compared to wild-type, kcat decreased compared to wild-type
Y378F
Km decreased compared to wild-type, kcat decreased compared to wild-type
E191D
catalytically inactive mutant is used for crystal structure determination
additional information
G464S/S465L
-
kcat values for DIBOA-glucoside decreased, Km (DIBOA-glucoside) increased, Km (DIMBOA-glucoside) decreased, kcat values for DIMBOA-glucoside increased
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
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
additional information
-
to study the mechanism of substrate specificity further, eight chimeric beta-glucosidases are constructed by replacing peptide sequences within the C-terminal region of Zea mays Glu1 with the homologous peptide sequences of Sorghum Dhr1 or vice versa. Replacing the peptide 466FAGFTERY473 of Zea mays Glu1 with the homologous peptide 462SSGYTERF469 of Sorghum Dhr1 or replacing the peptide 481NNNCTRYMKE490 in Glu1 with the homologous peptide 477ENGCERTMKR486 of Dhr1 is sufficient to confer to Zea mays Glu1 the ability to hydrolyze dhurrin
additional information
-
the N-terminal 25 residues of the mature TaGlu1a and TaGlu1b are replaced with the corresponding residues of Zea mays ZmGlu1, considering that this enzyme is known to exist as a dimer. The chimeric glucosidases (Zm-TaGlu1a and Zm-TaGlu1b) completely loses their ability to form a hexamer, which is confirmed by gel filtration chromatography
additional information
the N-terminal 25 residues of the mature TaGlu1a and TaGlu1b are replaced with the corresponding residues of Zea mays ZmGlu1, considering that this enzyme is known to exist as a dimer. The chimeric glucosidases (Zm-TaGlu1a and Zm-TaGlu1b) completely loses their ability to form a hexamer, which is confirmed by gel filtration chromatography
additional information
-
to study the mechanism of substrate specificity further, eight chimeric beta-glucosidases are constructed by replacing peptide sequences within the C-terminal region of Zea mays Glu1 with the homologous peptide sequences of Sorghum Dhr1 or vice versa. Replacing the peptide 466FAGFTERY473 of Zea mays Glu1 with the homologous peptide 462SSGYTERF469 of Sorghum Dhr1 or replacing the peptide 481NNNCTRYMKE490 in Glu1 with the homologous peptide 477ENGCERTMKR486 of Dhr1 is sufficient to confer to Zea mays Glu1 the ability to hydrolyze dhurrin
