Literature summary for 3.2.1.182 extracted from

  • Cicek, M.; Blanchard, D.; Bevan, D.R.; Esen, A.
    The aglycone specificity-determining sites are different in 2, 4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA)-glucosidase (Maize beta -glucosidase) and dhurrinase (Sorghum beta -glucosidase) (2000), J. Biol. Chem., 275, 20002-20011.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Zea mays
expressed in Escherichia coli as a His-tagged fusion protein Sorghum bicolor

Protein Variants

Protein Variants Comment Organism
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 Zea mays
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 Sorghum bicolor

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.2
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim16 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-495 of Sorghum Dhr1) Zea mays
0.2
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim16 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-495 of Sorghum Dhr1) Sorghum bicolor
0.104
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim21 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-473 of Sorghum Dhr1) Zea mays
0.104
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim21 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-473 of Sorghum Dhr1) Sorghum bicolor
0.098
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim22 (chimeric construct: parental Zea mays Glu1 with exchanged domain 481-490 of Sorghum Dhr1) Zea mays
0.098
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim22 (chimeric construct: parental Zea mays Glu1 with exchanged domain 481-490 of Sorghum Dhr1) Sorghum bicolor
0.098
-
DIMBOA-glucoside pH 5.8, 37°C, wild-type Zea mays
0.106
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim15 (chimeric construct: parental Zea mays Glu1 with exchanged domain 496-518 of Sorghum Dhr1) Zea mays
0.106
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim15 (chimeric construct: parental Zea mays Glu1 with exchanged domain 496-518 of Sorghum Dhr1) Sorghum bicolor
0.205
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim2 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-518 of Sorghum Dhr1) Zea mays
0.205
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim2 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-518 of Sorghum Dhr1) Sorghum bicolor

Organism

Organism UniProt Comment Textmining
Sorghum bicolor
-
-
-
Zea mays
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Zea mays
using Ni-NTA chromatography Sorghum bicolor

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
DIMBOA-glucoside + H2O
-
Zea mays DIMBOA + beta-D-glucose
-
?
DIMBOA-glucoside + H2O
-
Sorghum bicolor DIMBOA + beta-D-glucose
-
?

Synonyms

Synonyms Comment Organism
ZmGlu1
-
Zea mays
SbDhr1
-
Sorghum bicolor

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at 37°C, substrates: dhurrin and DIMBOA-glucoside Zea mays
37
-
assay at 37°C, substrates: dhurrin and DIMBOA-glucoside Sorghum bicolor
25
-
assay at 25°C, substrates: 2-nitrophenyl-beta-D-glucopyranoside and 4-nitrophenyl-beta-D-glucopyranoside Sorghum bicolor

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
43.33
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim2 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-518 of Sorghum Dhr1) Zea mays
43.33
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim2 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-518 of Sorghum Dhr1) Sorghum bicolor
19.54
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim16 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-495 of Sorghum Dhr1) Zea mays
19.54
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim16 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-495 of Sorghum Dhr1) Sorghum bicolor
25.67
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim22 (chimeric construct: parental Zea mays Glu1 with exchanged domain 481-490 of Sorghum Dhr1) Zea mays
25.67
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim22 (chimeric construct: parental Zea mays Glu1 with exchanged domain 481-490 of Sorghum Dhr1) Sorghum bicolor
29.16
-
DIMBOA-glucoside pH 5.8, 37°C, wild-type Zea mays
24.02
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim15 (chimeric construct: parental Zea mays Glu1 with exchanged domain 496-518 of Sorghum Dhr1) Zea mays
24.02
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim15 (chimeric construct: parental Zea mays Glu1 with exchanged domain 496-518 of Sorghum Dhr1) Sorghum bicolor
20.15
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim21 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-473 of Sorghum Dhr1) Zea mays
20.15
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim21 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-473 of Sorghum Dhr1) Sorghum bicolor

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.8
-
assay at Zea mays
5.8
-
assay at Sorghum bicolor

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
211
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim2 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-518 of Sorghum Dhr1) Zea mays
211
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim2 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-518 of Sorghum Dhr1) Sorghum bicolor
226
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim15 (chimeric construct: parental Zea mays Glu1 with exchanged domain 496-518 of Sorghum Dhr1) Zea mays
226
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim15 (chimeric construct: parental Zea mays Glu1 with exchanged domain 496-518 of Sorghum Dhr1) Sorghum bicolor
297.5
-
DIMBOA-glucoside pH 5.8, 37°C, wild-type Zea mays
97.7
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim16 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-495 of Sorghum Dhr1) Zea mays
97.7
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim16 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-495 of Sorghum Dhr1) Sorghum bicolor
195.2
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim21 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-473 of Sorghum Dhr1) Zea mays
195.2
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim21 (chimeric construct: parental Zea mays Glu1 with exchanged domain 466-473 of Sorghum Dhr1) Sorghum bicolor
261.9
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim22 (chimeric construct: parental Zea mays Glu1 with exchanged domain 481-490 of Sorghum Dhr1) Zea mays
261.9
-
DIMBOA-glucoside pH 5.8, 37°C, mutant Chim22 (chimeric construct: parental Zea mays Glu1 with exchanged domain 481-490 of Sorghum Dhr1) Sorghum bicolor