Literature summary for 4.2.2.3 extracted from

Mikami, B.; Ban, M.; Suzuki, S.; Yoon, H.J.; Miyake, O.; Yamasaki, M.; Ogura, K.; Maruyama, Y.; Hashimoto, W.; Murata, K.
Induced-fit motion of a lid loop involved in catalysis in alginate lyase A1-III (2012), Acta Crystallogr. Sect. D, 68, 1207-1216.

Cloned(Commentary)

Cloned (Comment)	Organism
sequence comparison of PL-5 alginate lyase, recombinant wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)pLysS	Sphingomonas sp.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant mutants H192A and Y246F complexed with substrate 4-deoxy-L-erythro-hex-4-ene-pyranosyluronate-(mannuronate)2-mannuronic acid, hanging drop vapour diffusion method, 10 mg/ml protein in solution is mixed with reservoir solution containing 24% w/v PEG 4000, 0.3 M ammonium acetate, 0.1 M sodium citrate pH 5.5, 20°C, 1 month, X-ray diffraction structure determination and analysis at 2.2 resolution, final models of the complex forms, which comprise two monomers (of 353 amino-acid residues each), 268-287 water molecules and two tetrasaccharide substrates	Sphingomonas sp.

Crystallization (Comment)

Organism

purified recombinant mutants H192A and Y246F complexed with substrate 4-deoxy-L-erythro-hex-4-ene-pyranosyluronate-(mannuronate)2-mannuronic acid, hanging drop vapour diffusion method, 10 mg/ml protein in solution is mixed with reservoir solution containing 24% w/v PEG 4000, 0.3 M ammonium acetate, 0.1 M sodium citrate pH 5.5, 20°C, 1 month, X-ray diffraction structure determination and analysis at 2.2 resolution, final models of the complex forms, which comprise two monomers (of 353 amino-acid residues each), 268-287 water molecules and two tetrasaccharide substrates

Sphingomonas sp.

Protein Variants

Protein Variants	Comment	Organism
G60A	site-directed active site mutagenesis, the mutant shows 41.4% reduced activity compared to the wild-type enzyme	Sphingomonas sp.
H192A	site-directed active site mutagenesis, almost inactive mutant	Sphingomonas sp.
M62P	site-directed active site mutagenesis, two components of M62P, intactM62P and nicked M62P, are found during purification of the mutant enzyme, the nicked form is inactive, the intact form shows 56% reduced activity compared to the wild-type enzyme	Sphingomonas sp.
R67A	site-directed active site mutagenesis, the mutant shows 92.5% reduced activity compared to the wild-type enzyme	Sphingomonas sp.
Y246F	site-directed active site mutagenesis, almost inactive mutant	Sphingomonas sp.
Y68F	site-directed active site mutagenesis, the mutant shows 95% reduced activity compared to the wild-type enzyme	Sphingomonas sp.
Y80F	site-directed active site mutagenesis, the mutant shows 47% reduced activity compared to the wild-type enzyme	Sphingomonas sp.

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	kinetics of wild-type and mutant enzymes, overview	Sphingomonas sp.

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Sphingomonas sp.	alginate lyase A1-III is a beta-D-mannuronosyl linkage-specific enzyme that acts on alginate tetrasaccharide as the minimum substrate and produces disaccharides and trisaccharides from alginate	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Sphingomonas sp.	Q75WP3	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)pLysS	Sphingomonas sp.

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-deoxy-L-erythro-hex-4-ene-pyranosyluronate-(mannuronate)2-mannuronic acid	bound in the active cleft at subsites -3 to +1, the glycosidic linkage is cleaved existed between subsites -1 and +1	Sphingomonas sp.	?	-	?
additional information	alginate lyase A1-III is a beta-D-mannuronosyl linkage-specific enzyme that acts on alginate tetrasaccharide as the minimum substrate and produces disaccharides and trisaccharides from alginate	Sphingomonas sp.	?	-	?

Synonyms

Synonyms	Comment	Organism
alginate lyase A1-III	-	Sphingomonas sp.
PL-5 alginate lyase	-	Sphingomonas sp.

General Information

General Information	Comment	Organism
evolution	sequence comparison of the PL-5 alginate lyase	Sphingomonas sp.
additional information	structure-fucntion analysis, superposition of the open and closed lid loops suggests that the conformational change results in near-rigid-body motion, The open-closed movement of the lid loop decreases the accessible surface area by covering the active-site cleft., overview	Sphingomonas sp.