Literature summary for 3.2.1.21 extracted from

Verdoucq, L.; Moriniere, J.; Bevan, D.R.; Esen, A.; Vasella, A.; Henrissat, B.; Czjze, M.
Structural determinants of substrate specificity in family 1 beta-glucosidases: novel insights from the crystal structure of sorghum dhurrinase-1, a plant beta-glucosidase with strict specificity, in complex with its natural substrate (2004), J. Biol. Chem., 279, 31796-31803.

Search for more literature for 3.2.1.21

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression of Dhr1 and mutant E189D in Escherichia coli as His-tagged proteins	Sorghum bicolor

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant Glu1 mutant E191D in complex with cyclic inhibitor glucoetrazole, the crystallization solution is miced with 0.005 ml glycerol and 0.003 ml of inhibitor solution, X-ray diffraction structure determination and analysis at 1.95 A resolution	Zea mays
wild-type Dhr1, and Dhr1 mutant E189D in complex with dhurrin, hanging drop vapour diffusion method, 0.002 ml of 8 mg/ml protein in 20 mM Tris, pH 7.0, mixed with an equal volume of reservoir solution containing 0.1 M HEPES, pH 7.5, 2.0 M ammonium sulfate, and 50 mM NaCl, X-ray diffraction structure determination and analysis at 1.9-2.0 A resolution	Sorghum bicolor

Protein Variants

Protein Variants	Comment	Organism
E189D	substrate binding structure analysis from crystal structure	Sorghum bicolor
E191D	substrate binding structure analysis from crystal structure	Zea mays
E191D/F198V	inactive mutant, F198 substitution causes a rearrangement of residues F205, F466, and E464 involved in aglycone binding	Zea mays

Inhibitors

Inhibitors	Comment	Organism	Structure
cyclic glucotetrazole	mimics the 4H3 transition state intermediate	Sorghum bicolor
cyclic glucotetrazole	mimics the 4H3 transition state intermediate	Zea mays

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one + H2O	Zea mays	-	2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one + beta-D-glucose	-	?
dhurrin + H2O	Sorghum bicolor	i.e. 4-hydroxy-(S)-mandelonitrile-beta-D-glucoside, Dhr1 is strictly specific for this substrate	4-hydroxy-(S)-mandelonitrile + beta-D-glucose	-	?

Organism

Organism	UniProt	Comment	Textmining
Sorghum bicolor	-	isozyme Dhr1	-
Zea mays	P49235	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged Dhr1 and mutant E189D from Escherichia coli by nickel affinity chromatography	Sorghum bicolor

Reaction

Reaction	Comment	Organism	Reaction ID
celloheptaose + 6 H2O = 7 beta-D-glucose	structural basis for substrate specificity, residues N259, F261, and S462 are important, substrate binding mode and structure	Sorghum bicolor	a
celloheptaose + 6 H2O = 7 beta-D-glucose	substrate binding mode and structure	Zea mays	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one + H2O	-	Zea mays	2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one + beta-D-glucose	-	?
2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one + H2O	i.e. DIMBOA-glucoside	Zea mays	2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one + beta-D-glucose	-	?
dhurrin + H2O	i.e. 4-hydroxy-(S)-mandelonitrile-beta-D-glucoside, Dhr1 is strictly specific for this substrate	Sorghum bicolor	4-hydroxy-(S)-mandelonitrile + beta-D-glucose	-	?
additional information	Glu1 shows broad substrate specificity in vitro	Zea mays	?	-	?

Synonyms

Synonyms	Comment	Organism
Dhr1	-	Sorghum bicolor
dhurrinase-1	-	Sorghum bicolor
Glu1	-	Zea mays

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Release 2023.1 (January 2023)