Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Xanthomonas axonopodis |
Crystallization (Comment) | Organism |
---|---|
purified mutant E322Q in complex with Tris or sucrose, E322Q SUH is crystallized by hanging-drop vapor-diffusion method at 22°C in a crystallization buffer consisting of 0.1 M HEPES, pH 6.5, 0.2 M calcium acetate, 16% PEG 3000, and 5 mM DTT, with 0.1 M caesium chloride, and crystallization of SeMet SUH sitting-drop method at 22°C in a crystallization buffer consisting of 0.1 M HEPES, pH 6.5, 0.2 M calcium acetate, 16% PEG 3000, and 5 mM DTT, with 0.1 M guanidine chloride, X-ray diffraction structure determination and analysis at 1.8 A resolution | Xanthomonas axonopodis |
Protein Variants | Comment | Organism |
---|---|---|
E322Q | site-directed mutagenesis, a catalytically inactive Xag SUH mutant | Xanthomonas axonopodis |
G219R | site-directed mutagenesis, the mutant shows increased catalytic activity compared to the wild-type enzyme | Xanthomonas axonopodis |
G219R/G444R | site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme | Xanthomonas axonopodis |
G219R/L414R | site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme | Xanthomonas axonopodis |
G444R | site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme | Xanthomonas axonopodis |
L414R | site-directed mutagenesis, the mutant shows slightly reduced catalytic activity compared to the wild-type enzyme | Xanthomonas axonopodis |
L414R/G444R | site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme | Xanthomonas axonopodis |
additional information | conformational changes in the SUH active site du to mutational alterations, overview | Xanthomonas axonopodis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.3 | - |
sucrose | recombinant mutant L414R/G444R | Xanthomonas axonopodis | |
1.34 | - |
sucrose | recombinant mutant G444R | Xanthomonas axonopodis | |
1.73 | - |
sucrose | recombinant mutant G219R/G444R | Xanthomonas axonopodis | |
2.24 | - |
sucrose | recombinant wild-type enzyme | Xanthomonas axonopodis | |
2.48 | - |
sucrose | recombinant mutant G219R | Xanthomonas axonopodis | |
2.97 | - |
sucrose | recombinant mutant L414R | Xanthomonas axonopodis | |
3.6 | - |
sucrose | recombinant mutant G219R/L4141R | Xanthomonas axonopodis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Xanthomonas axonopodis | the enzyme is identified as NpAS, i.e. Neisseria polysaccharea amylosucrase, homolog, involved in regulation of the utilization of plant sucrose in phytopathogenic bacteria. But the enzyme is exclusively a hydrolase and not a glucosyltransferase and is termed sucrose hydrolase, SUH, overview | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Xanthomonas axonopodis | Q6UVM5 | sucrose hydrolase; pv. glycines | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme is identified as NpAS, i.e. Neisseria polysaccharea amylosucrase, homolog, involved in regulation of the utilization of plant sucrose in phytopathogenic bacteria. But the enzyme is exclusively a hydrolase and not a glucosyltransferase and is termed sucrose hydrolase, SUH, overview | Xanthomonas axonopodis | ? | - |
? | |
sucrose + glucan | SUH active site structure analysis, overview | Xanthomonas axonopodis | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
sucrose hydrolase | - |
Xanthomonas axonopodis |
SUH | - |
Xanthomonas axonopodis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
12.34 | - |
sucrose | recombinant mutant G444R | Xanthomonas axonopodis | |
12.49 | - |
sucrose | recombinant mutant L414R/G444R | Xanthomonas axonopodis | |
12.77 | - |
sucrose | recombinant mutant G219R/G444R | Xanthomonas axonopodis | |
60.64 | - |
sucrose | recombinant mutant G219R/L4141R | Xanthomonas axonopodis | |
66.45 | - |
sucrose | recombinant wild-type enzyme | Xanthomonas axonopodis | |
78.01 | - |
sucrose | recombinant mutant L414R | Xanthomonas axonopodis | |
107 | - |
sucrose | recombinant mutant G219R | Xanthomonas axonopodis |