Literature summary for 3.2.1.48 extracted from

Kim, M.I.; Kim, H.S.; Jung, J.; Rhee, S.
Crystal structures and mutagenesis of sucrose hydrolase from Xanthomonas axonopodis pv. glycines: insight into the exclusively hydrolytic amylosucrase fold (2008), J. Mol. Biol., 380, 636-647.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Xanthomonas axonopodis

Crystallization (Commentary)

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

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

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 [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Xanthomonas axonopodis	Q6UVM5	sucrose hydrolase; pv. glycines	-

Substrates and Products (Substrate)

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

Synonyms	Comment	Organism
sucrose hydrolase	-	Xanthomonas axonopodis
SUH	-	Xanthomonas axonopodis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
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