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Literature summary extracted from

  • Labourel, A.; Jam, M.; Jeudy, A.; Hehemann, J.H.; Czjzek, M.; Michel, G.
    The beta-glucanase ZgLamA from Zobellia galactanivorans evolved a bent active site adapted for efficient degradation of algal laminarin (2014), J. Biol. Chem., 289, 2027-2042.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.39 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
3.2.1.73 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 (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.39 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
3.2.1.73 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

EC Number Protein Variants Comment Organism
3.2.1.39 E269S site-directed mutagenesis, nucleophile replacement, inactive mutant Zobellia galactanivorans
3.2.1.73 E269S site-directed mutagenesis, nucleophile replacement, inactive mutant Zobellia galactanivorans

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.39 laminarin + H2O Zobellia galactanivorans
-
 ?
-
 ?
3.2.1.39 laminarin + H2O Zobellia galactanivorans DSM 12802
-
 ?
-
 ?
3.2.1.73 laminarin + H2O Zobellia galactanivorans
-
 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.39 Zobellia galactanivorans G0L5X4 gene Zobellia_2431
-
3.2.1.39 Zobellia galactanivorans DSM 12802 G0L5X4 gene Zobellia_2431
-
3.2.1.73 Zobellia galactanivorans G0L5X4 gene Zobellia_2431
-
3.2.1.73 Zobellia galactanivorans DSM 12802 G0L5X4 gene Zobellia_2431
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.39 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and gel filtration Zobellia galactanivorans
3.2.1.73 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 and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.39 Glc-beta-(1->4)-Glc-beta-(1->3)-Glc-beta-(1->4)-Glc + H2O
-
 Zobellia galactanivorans D-glucose + Glc-beta-(1->4)-Glc-beta-(1->3)-Glc containing one beta-1,4-linkage separated by one beta-1,3-linkage, i.e. G4G3G, which is not further degraded ?
3.2.1.39 Glc-beta-(1->4)-Glc-beta-(1->3)-Glc-beta-(1->4)-Glc + H2O
-
 Zobellia galactanivorans DSM 12802 D-glucose + Glc-beta-(1->4)-Glc-beta-(1->3)-Glc containing one beta-1,4-linkage separated by one beta-1,3-linkage, i.e. G4G3G, which is not further degraded ?
3.2.1.39 laminarin + H2O
-
 Zobellia galactanivorans ?
-
 ?
3.2.1.39 laminarin + H2O the wild-type enzyme displays a bent topology adapted to the binding of helical-shaped laminarin Zobellia galactanivorans ?
-
 ?
3.2.1.39 laminarin + H2O
-
 Zobellia galactanivorans DSM 12802 ?
-
 ?
3.2.1.39 laminarin + H2O the wild-type enzyme displays a bent topology adapted to the binding of helical-shaped laminarin Zobellia galactanivorans DSM 12802 ?
-
 ?
3.2.1.39 laminaritriose + H2O the smallest oligosaccharide that can be degraded by the enzyme Zobellia galactanivorans D-glucose + laminaribiose
-
 ?
3.2.1.39 laminaritriose + H2O the smallest oligosaccharide that can be degraded by the enzyme Zobellia galactanivorans DSM 12802 D-glucose + laminaribiose
-
 ?
3.2.1.39 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. The activity towards beta-1,4-linkages is much lower than towards beta-1,3-linkages. Enzyme-substrate complex structure analysis using wild-type and mutant enzymes with glucan oligomers, binding structure and substrate recognition, overview Zobellia galactanivorans ?
-
 ?
3.2.1.39 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. The activity towards beta-1,4-linkages is much lower than towards beta-1,3-linkages. Enzyme-substrate complex structure analysis using wild-type and mutant enzymes with glucan oligomers, binding structure and substrate recognition, overview Zobellia galactanivorans DSM 12802 ?
-
 ?
3.2.1.73 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 ?
3.2.1.73 laminarin + H2O
-
 Zobellia galactanivorans ?
-
 ?
3.2.1.73 laminarin + H2O the wild-type enzyme displays a bent topology adapted to the binding of helical-shaped laminarin Zobellia galactanivorans ?
-
 ?
3.2.1.73 laminaritriose + H2O the smallest oligosaccharide that can be degraded by the enzyme Zobellia galactanivorans D-glucose + laminaribiose
-
 ?
3.2.1.73 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

EC Number Synonyms Comment Organism
3.2.1.39 beta-glucanase
-
 Zobellia galactanivorans
3.2.1.39 LamA
-
 Zobellia galactanivorans
3.2.1.39 laminarinase
-
 Zobellia galactanivorans
3.2.1.39 ZgLamA
-
 Zobellia galactanivorans
3.2.1.73 beta-glucanase
-
 Zobellia galactanivorans
3.2.1.73 LamA
-
 Zobellia galactanivorans
3.2.1.73 laminarinase
-
 Zobellia galactanivorans
3.2.1.73 ZgLamA
-
 Zobellia galactanivorans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.39 37 40 assay at Zobellia galactanivorans
3.2.1.73 37 40 assay at Zobellia galactanivorans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.39 8.5
-
 assay at Zobellia galactanivorans
3.2.1.73 8.5
-
 assay at Zobellia galactanivorans

General Information

EC Number General Information Comment Organism
3.2.1.39 evolution the enzyme belongs to the glycohydrolase family GH16 Zobellia galactanivorans
3.2.1.39 additional information involvement of Glu269 in the catalytic machinery of the enzyme Zobellia galactanivorans
3.2.1.73 evolution the enzyme belongs to the glycohydrolase family GH16 Zobellia galactanivorans
3.2.1.73 additional information involvement of Glu269 in the catalytic machinery of the enzyme Zobellia galactanivorans