Cloned (Comment) | Organism |
---|---|
gene Zobellia_2431, DNA and amino acid sequence determination and analysis, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Zobellia galactanivorans |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged wild-type and mutant enzymes, hanging drop vapour diffusion method, mixing of 0.002 ml of 10 mg/ml protein with 0.002 ml reservoir solution containing 24% PEG 3350 and 100 mM sodium citrate, pH 5.2, 4°C, crystals of enzyme mutant E269S in complex with laminarin oligosaccharides are obtained from 0.002 ml of 13.3 mg/ml protein and 5 mM of purified hexasaccharides,with 0.001 ml of reservoir solution containing 100 mM sodium malonate, imidazole, and boric acid (MIB-buffer), pH 4.0, and 19% of PEG 1500 in hanging drops at 20°C. Single crystals of mutant E269S in complex with MLG trisaccharides are obtained from mixing 0.002 ml 11.7 mg/ml of protein, 0.04% w/v of MLG degradation products with 0.001 ml of reservoir solution containing 100 mM MIB buffer, pH 4.0, 17% of PEG 1500, and 10% of glycerol in hanging drops at 12°C, X-ray diffraction structure determination and analysis at 1.13-1.45 A resolution, molecular replacement method | Zobellia galactanivorans |
Protein Variants | Comment | Organism |
---|---|---|
E269S | site-directed mutagenesis, nucleophile replacement, inactive mutant | Zobellia galactanivorans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
laminarin + H2O | Zobellia galactanivorans | - |
? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Zobellia galactanivorans | G0L5X4 | gene Zobellia_2431 | - |
Zobellia galactanivorans DSM 12802 | G0L5X4 | gene Zobellia_2431 | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and gel filtration | Zobellia galactanivorans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glucan tetrasaccharide + H2O | containing two beta-1,4-linkages separated by one beta-1,3-linkage, i.e. G4G3G4G | Zobellia galactanivorans | D-glucose + glucan trisaccharide | containing one beta-1,4-linkage separated by one beta-1,3-linkage, i.e. G4G3G, which is not further degraded | ? | |
laminarin + H2O | - |
Zobellia galactanivorans | ? | - |
? | |
laminarin + H2O | the wild-type enzyme displays a bent topology adapted to the binding of helical-shaped laminarin | Zobellia galactanivorans | ? | - |
? | |
laminaritriose + H2O | the smallest oligosaccharide that can be degraded by the enzyme | Zobellia galactanivorans | D-glucose + laminaribiose | - |
? | |
additional information | the enzyme specifically cleaves beta-1,4-linkages next to beta-1,3-linkages and is active on linear 1,3-beta-D-glucans with 3-6 monomers and on a glucan tetrasaccharide containing two beta-1,4-linkages separated by one beta-1,3-linkage. Enzyme-substrate complex structure analysis using wild-type and mutant enzymes with glucan oligomers, binding structure and substrate recognition, overview | Zobellia galactanivorans | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
beta-glucanase | - |
Zobellia galactanivorans |
LamA | - |
Zobellia galactanivorans |
laminarinase | - |
Zobellia galactanivorans |
ZgLamA | - |
Zobellia galactanivorans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | 40 | assay at | Zobellia galactanivorans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Zobellia galactanivorans |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycohydrolase family GH16 | Zobellia galactanivorans |
additional information | involvement of Glu269 in the catalytic machinery of the enzyme | Zobellia galactanivorans |