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 (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 | 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 Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of wild-type and mutant enzymes, overview | Sphingomonas sp. |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Sphingomonas sp. | Q75WP3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)pLysS | Sphingomonas sp. |
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 | Comment | Organism |
---|---|---|
alginate lyase A1-III | - |
Sphingomonas sp. |
PL-5 alginate lyase | - |
Sphingomonas sp. |
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. |