Cloned (Comment) | Organism |
---|---|
expressed as a His-tagged fusion protein | Sphingomonas sp. |
Protein Variants | Comment | Organism |
---|---|---|
H191N/Y284F | crystal structure of mutant H191N/Y284F complexed with a tetrasaccharide bound at subsites -1 to +3 suggests that Gln189 functions as a neutralizer for the substrate carboxyl group, His191 as a general base, and Tyr284 as a general acid | Sphingomonas sp. |
N141C/N199C | almost all molecules of N141C/N199C form a disulfide bond between Cys141 and Cys199. Crystal structure of N141C/N199C is determined at 2.1 A resolution by X-ray crystallography. Mutant has a glove-like beta- sandwich structure composed of four short alpha-helices and two beta-sheets. Two sulfate ions derived from the crystallization solution are accommodated at subsites +1 and +3. The loops of the mutant adopt the closed form. A disulfide bond between Cys141 and Cys199 forms in the absence of reducing agents | Sphingomonas sp. |
N141C/N199C | Km (mg/ml) (alginate): 0.32 (N141C/N199C + 0.5 mM DTT), 0.14 (N141C/N199C), Vmax (U/mg) (alginate): 7.1 (N141C/N199C + 0.5 mM DTT), 0.63 (N141C/N199C) | Sphingomonas sp. |
Y284F | molecular activity is significantly lower than that of wild-type enzyme. GGG- and MMG bound Y284F crystal structure are refined at 1.65 A and 1.55 A resolution. Mutant enzyme interacts appropriately with substrate hydroxyl groups at subsites +1 and +2 and accommodates alpha-L-guluronate or beta-D-mannuronate, while substrate carboxyl groups are strictly recognized by specific residues | Sphingomonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Km (mg/ml) (alginate): 0.018 (wild-type), 0.32 (mutant N141C/N199C + 0.5 mM DTT), 0.14 (mutant N141C/N199C) | Sphingomonas sp. | |
additional information | - |
additional information | Vmax (U/mg) (alginate): 42.9 (wild-type), 7.1 (mutant N141C/N199C + 0.5 mM DTT), 0.63 (mutant N141C/N199C) | Sphingomonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sphingomonas sp. | - |
- |
- |
Purification (Comment) | Organism |
---|---|
using Ni-NTA chromatography | Sphingomonas sp. |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
R2-beta-D-mannuronic acid-R1 = R2-OH + 4-deoxy-alpha-L-erythro-hex-4-enopyranuronosyl-R1 | A1-II with a glove-like beta-sandwich as a basic scaffold forms a cleft covered with two lid loops (L1/L2). Loop flexibility for substrate binding and structural determinants for broad substrate recognition and catalytic reaction is shown. The two loops associate mutually over the cleft through the formation of a hydrogen bond between their edges (Asn141 and Asn199) | Sphingomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
alginate | - |
Sphingomonas sp. | unsaturated algino-oligosaccharides | - |
? |
Synonyms | Comment | Organism |
---|---|---|
A1 alginate lyase | - |
Sphingomonas sp. |
A1-II | - |
Sphingomonas sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Sphingomonas sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Sphingomonas sp. |