Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant enzymes in Escherichia coli | Secale cereale |
isozyme TaGlu1a, DNA and amino acid sequence determination and analysis, isozyme expression profile, expression of wild-type and mutant isozyme TaGlu1a in Escherichia coli | Triticum aestivum |
isozyme TaGlu1b, DNA and amino acid sequence determination and analysis, isozyme expression profile, expression in Escherichia coli | Triticum aestivum |
isozyme TaGlu1c, DNA and amino acid sequence determination and analysis, isozyme expression profile, expression in Escherichia coli | Triticum aestivum |
Crystallization (Comment) | Organism |
---|---|
purified recombinant isozyme TaGlu1a, X-ray diffraction structure determination and analysis at 1.8 A resolution | Triticum aestivum |
Protein Variants | Comment | Organism |
---|---|---|
E191A | site-directed mutagenesis, inactive mutant | Secale cereale |
E191A | site-directed mutagenesis, inactive TaGlu1a mutant | Triticum aestivum |
E407A | site-directed mutagenesis, inactive mutant | Secale cereale |
E407A | site-directed mutagenesis, inactive TaGlu1a mutant | Triticum aestivum |
F198A | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme | Secale cereale |
F198A | site-directed mutagenesis, the mutant shows 24-28fold reduced activity with substrates 2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one and 2-O-beta-D-glucopyranosyl-4-hydroxy-7-demethoxy-1,4-benzoxazin-3-one, respectively | Secale cereale |
F198A | site-directed mutagenesis, the TaGlu1a mutant shows reduced activity with substrates 2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one and 2-O-beta-D-glucopyranosyl-4-hydroxy-7-demethoxy-1,4-benzoxazin-3-one with highly incresed Km values, respectively | Triticum aestivum |
F471Y | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme | Secale cereale |
G464F | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme | Secale cereale |
G464S | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme | Secale cereale |
G464S/S465L | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme | Secale cereale |
S465L | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme | Secale cereale |
Y378A | site-directed mutagenesis, the mutant shows 300-380% increased catalytic eficiency with substrates 2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one and 2-O-beta-D-glucopyranosyl-4-hydroxy-7-demethoxy-1,4-benzoxazin-3-one, respectively | Secale cereale |
Y378F | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme | Secale cereale |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
additional information | - |
formation of both homo- and heterohexamers by the isozyme | Triticum aestivum |
additional information | - |
formation of both homo- and heterohexamers by the isozymes | Triticum aestivum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Secale cereale | Q9FYS3 | isozyme TaGlu1a | - |
Triticum aestivum | Q1XH04 | isozyme TaGlu1c; isozyme TaGlu1c | - |
Triticum aestivum | Q1XH05 | isozyme TaGlu1b; isozyme TaGlu1b | - |
Triticum aestivum | Q1XIR9 | isozyme TaGlu1a; isozyme TaGlu1a | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli by metal affinity chromatography and gel filtration to homogeneity | Secale cereale |
recombinant wild-type and mutant isozyme TaGlu1a from Escherichia coli by metal affinity chromatography and gel filtration to homogeneity | Triticum aestivum |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
shoot | young, isozyme TaGlu1a, isozyme expression profile, overview | Triticum aestivum | - |
shoot | young, isozyme TaGlu1b, isozyme expression profile, overview | Triticum aestivum | - |
shoot | young, isozyme TaGlu1c, isozyme expression profile, overview | Triticum aestivum | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
245.9 | - |
purified native enzyme | Triticum aestivum |
311.9 | - |
purified recombinant His-tagged isozyme TaGlu1a | Triticum aestivum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-O-beta-D-glucopyranosyl-4-hydroxy-7-demethoxy-1,4-benzoxazin-3-one + H2O | - |
Secale cereale | 2,4-dihydroxy-7-demethoxy-1,4-benzoxazin-3-one + beta-D-glucose | - |
? | |
2-O-beta-D-glucopyranosyl-4-hydroxy-7-demethoxy-1,4-benzoxazin-3-one + H2O | - |
Triticum aestivum | 2,4-dihydroxy-7-demethoxy-1,4-benzoxazin-3-one + beta-D-glucose | - |
? | |
2-O-beta-D-glucopyranosyl-4-hydroxy-7-demethoxy-1,4-benzoxazin-3-one + H2O | Ser464 and Leu465 are critical in the substrate specificity for the substrate of isozyme TaGlu1a | Triticum aestivum | 2,4-dihydroxy-7-demethoxy-1,4-benzoxazin-3-one + beta-D-glucose | - |
? | |
2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one + H2O | - |
Secale cereale | 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one + beta-D-glucose | - |
? | |
2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one + H2O | - |
Triticum aestivum | 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one + beta-D-glucose | - |
? | |
2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one + H2O | Ser464 and Leu465 are critical in the substrate specificity for the substrate of isozyme TaGlu1a | Triticum aestivum | 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one + beta-D-glucose | - |
? | |
4-nitrophenyl beta-D-glucopyranoside + H2O | - |
Triticum aestivum | 4-nitrophenol + beta-D-glucose | - |
? | |
4-nitrophenyl beta-D-glucopyranoside + H2O | - |
Secale cereale | 4-nitrophenol + beta-D-glucose | - |
? | |
additional information | the wheat beta-D-glucosidases hydrolyze hydroxamic acid-glucose conjugates, overview | Triticum aestivum | ? | - |
? | |
additional information | the wheat beta-D-glucosidases hydrolyze hydroxamic acid-glucose conjugates, the aromatic side chain at position 378 of isozyme TaGlu1a, which is located at the entrance to the catalytic center, plays an important role in substrate binding, overview | Triticum aestivum | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | 6 * 64085.41, recombinant His-tagged TaGlu1b, SDS-PAGE, gel filtration, and mass spectrometry | Triticum aestivum |
hexamer | 6 * 64141.25, recombinant His-tagged isozyme TaGlu1a, SDS-PAGE, gel filtration, and mass spectrometry | Triticum aestivum |
hexamer | formation of both homo- and heterohexamers by the isozymes, the hexameric form is the active form, smaller oligomers or monomers are inactive, overview | Triticum aestivum |
More | three-dimensional structure modeling | Secale cereale |
More | formation of both homo- and heterohexamers by the isozymes, the hexameric form is the active form, smaller oligomers or monomers are inactive, overview | Triticum aestivum |
More | formation of both homo- and heterohexamers by the isozymes, the hexameric form is the active form, smaller oligomers or monomers are inactive, the N-terminal region of isozyme TaGlu1a is located at the dimer-dimer interface and plays a crucial role in hexamer formation, primary structure of the aglycone binding site, overview | Triticum aestivum |
Synonyms | Comment | Organism |
---|---|---|
beta-D-glucosidase | - |
Triticum aestivum |
beta-D-glucosidase | - |
Secale cereale |