Literature summary for 4.2.2.3 extracted from

Ochiai, A.; Yamasaki, M.; Mikami, B.; Hashimoto, W.; Murata, K.
Crystal structure of exotype alginate lyase Atu3025 from Agrobacterium tumefaciens (2010), J. Biol. Chem., 285, 24519-24528.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli B834(DE3) cells	Agrobacterium tumefaciens
expression in Escherichia coli	Agrobacterium tumefaciens

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystal structures of native protein and its inactive mutant H531A in complex with alginate trisaccharide, at 2.10 and 2.99 A resolutions with final R-factors of 18.3 and 19.9%, respectively. The enzyme is comprised of an alpha/alpha-barrel plus anti-parallel beta-sheet as a basic scaffold. His311 and Tyr365 are the catalytic base and acid, respectively. A short alpha-helix in the central alpha/alpha-barrel domain and a conformational change at the interface between the central and C-terminal domains are essential for the exolytic mode of action	Agrobacterium tumefaciens
wild type and mutant enzyme H531A in complex with alginate trisaccharide, sitting drop vapor diffusion method, using 80 mM Tris-HCl (pH 8.5), 24% (w/v) polyethylene glycol 4,000, 0.16 M magnesium chloride, and 20% (v/v) glycerol	Agrobacterium tumefaciens

Crystallization (Comment)

Organism

crystal structures of native protein and its inactive mutant H531A in complex with alginate trisaccharide, at 2.10 and 2.99 A resolutions with final R-factors of 18.3 and 19.9%, respectively. The enzyme is comprised of an alpha/alpha-barrel plus anti-parallel beta-sheet as a basic scaffold. His311 and Tyr365 are the catalytic base and acid, respectively. A short alpha-helix in the central alpha/alpha-barrel domain and a conformational change at the interface between the central and C-terminal domains are essential for the exolytic mode of action

Agrobacterium tumefaciens

wild type and mutant enzyme H531A in complex with alginate trisaccharide, sitting drop vapor diffusion method, using 80 mM Tris-HCl (pH 8.5), 24% (w/v) polyethylene glycol 4,000, 0.16 M magnesium chloride, and 20% (v/v) glycerol

Agrobacterium tumefaciens

Protein Variants

Protein Variants	Comment	Organism
H311A	0.3% of wild-type activity	Agrobacterium tumefaciens
H531A	inactive	Agrobacterium tumefaciens
R199A	4.3% of wild-type activity	Agrobacterium tumefaciens
W467A	0.45% of wild-type activity	Agrobacterium tumefaciens
Y365F	0.011% of wild-type activity	Agrobacterium tumefaciens

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
87870	-	calculated from amino acid sequence	Agrobacterium tumefaciens

Organism

Organism	UniProt	Comment	Textmining
Agrobacterium tumefaciens	A9CEJ9	-	-
Agrobacterium tumefaciens	A9CEJ9	polysaccharide lyase family 15	-

Purification (Commentary)

Purification (Comment)	Organism
DEAE-Toyopearl 650M column chromatography, Q Sepharose column chromatography, and Superdex 200 gel filtration	Agrobacterium tumefaciens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
1.14	-	wild-type, pH 7.5, 30°C	Agrobacterium tumefaciens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
alginate	-	Agrobacterium tumefaciens	?	-	?
alginate	ATU3025 acts on alginate polysaccharides and oligosaccharides exolytically and releases unsaturated monosaccharides from the substrate terminal	Agrobacterium tumefaciens	unsaturated algino-monosaccharides	-	?

Synonyms

Synonyms	Comment	Organism
Atu3025	member of the polysaccharide lyase family 15	Agrobacterium tumefaciens
exotype alginate lyase	-	Agrobacterium tumefaciens