EC Number | Cloned (Comment) | Organism |
---|---|---|
4.2.2.3 | phylogenetic analysis, recombinant expression of His-tagged AlyA5 in Escherichia coli strain BL21(DE3) | Zobellia galactanivorans |
4.2.2.11 | phylogenetic analysis, recombinant expression of His-tagged AlyA1PL7 in Escherichia coli strain BL21(DE3) | Zobellia galactanivorans |
4.2.2.11 | phylogenetic analysis, recombinant expression of His-tagged AlyA5 in Escherichia coli strain BL21(DE3) | Zobellia galactanivorans |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
4.2.2.3 | purified recombinant full-length dimeric AlyA5, hanging drop vapor diffusion method, 0.002 ml of 7.7 mg/ml protein solution with 0.1 mg/ml oligoglucuronate, are mixed with 0.001 ml of reservoir solution containing PEG 3350 and 0.2 M sodium/potassium tartrate, and equilibration against 0.2 ml reservoir solution, 21°C, screening and method optimization, X-ray diffraction structure determination and analysis at 1.75 A resolution | Zobellia galactanivorans |
4.2.2.11 | purified recombinant catalytic domain of AlyA1PL7, hanging drop vapor diffusion method, 0.002 ml of 11.3 mg/ml protein solution with 0.1 mg/ml oligoguluronate, are mixed with 0.001 ml of reservoir solution containing 0.2 M KSCN and 28% PEG-MME 2000, and equilibration against 0.5 ml reservoir solution, 21°C, screening and method optimization, X-ray diffraction structure determination and analysis at 1.43 A resolution | Zobellia galactanivorans |
4.2.2.11 | purified recombinant full-length dimeric AlyA5, hanging drop vapor diffusion method, 0.002 ml of 7.7 mg/ml protein solution with 0.1 mg/ml oligoglucuronate, are mixed with 0.001 ml of reservoir solution containing PEG 3350 and 0.2 M sodium/potassium tartrate, and equilibration against 02 ml reservoir solution, 21°C, screening and method optimization, X-ray diffraction structure determination and analysis at 1.75 A resolution | Zobellia galactanivorans |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
4.2.2.11 | EDTA | 90% inhibition | Zobellia galactanivorans | |
4.2.2.11 | EGTA | 95% inhibition | Zobellia galactanivorans |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.2.2.3 | additional information | - |
additional information | dimeric AlyA5 shows two-phase kinetics with alginate, overview | Zobellia galactanivorans | |
4.2.2.11 | additional information | - |
additional information | dimeric AlyA5 shows two-phase kinetics with alginate, overview | Zobellia galactanivorans | |
4.2.2.11 | 1.753 | - |
sodium alginate | with L-guluronate content of 66.7%, pH 7.5, 30°C, recombinant enzyme | Zobellia galactanivorans | |
4.2.2.11 | 3.087 | - |
sodium alginate | with L-guluronate content of 52.6%, pH 7.5, 30°C, recombinant enzyme | Zobellia galactanivorans | |
4.2.2.11 | 6.18 | - |
sodium alginate | with L-guluronate content of 33.3%, pH 7.5, 30°C, recombinant enzyme | Zobellia galactanivorans |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
4.2.2.3 | Ca2+ | at least 2 mM Ca2+ in the reaction mixture is essential for the activity | Zobellia galactanivorans | |
4.2.2.11 | Ca2+ | at least 2 mM Ca2+ in the reaction mixture is essential for the activity | Zobellia galactanivorans | |
4.2.2.11 | Ca2+ | requires a divalent cation for activity, interaction between the enzyme and divalent cations takes place during substrate fixation and not before | Zobellia galactanivorans | |
4.2.2.11 | NaCl | activates, best at 150-200 mM | Zobellia galactanivorans |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
4.2.2.3 | 38300 | - |
monomeric recombinant His-tagged enzyme, gel filtration | Zobellia galactanivorans |
4.2.2.3 | 42000 | - |
x * 42000, recombinant His-tagged enzyme, SDS-PAGE | Zobellia galactanivorans |
4.2.2.3 | 69500 | - |
dimeric recombinant His-tagged enzyme, gel filtration | Zobellia galactanivorans |
4.2.2.11 | 38300 | - |
monomeric recombinant His-tagged enzyme, gel filtration | Zobellia galactanivorans |
4.2.2.11 | 42000 | - |
x * 42000, recombinant His-tagged enzyme, SDS-PAGE | Zobellia galactanivorans |
4.2.2.11 | 69500 | - |
dimeric recombinant His-tagged enzyme, gel filtration | Zobellia galactanivorans |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
4.2.2.3 | sodium alginate | Zobellia galactanivorans | AlyA5 cleaves unsaturated units, alpha-L-guluronate or beta-D-manuronate residues, at the nonreducing end of oligo-alginates in an exolytic fashion, cf. EC 4.2.2.11 | ? | - |
? | |
4.2.2.3 | sodium alginate | Zobellia galactanivorans DSM 12802 | AlyA5 cleaves unsaturated units, alpha-L-guluronate or beta-D-manuronate residues, at the nonreducing end of oligo-alginates in an exolytic fashion, cf. EC 4.2.2.11 | ? | - |
? | |
4.2.2.11 | sodium alginate | Zobellia galactanivorans | AlyA1PL7 is an endolytic guluronate lyase that preferentially cleaves guluronate stretches | ? | - |
? | |
4.2.2.11 | sodium alginate | Zobellia galactanivorans | AlyA5 cleaves unsaturated units, alpha-L-guluronate or beta-D-manuronate residues, at the nonreducing end of oligo-alginates in an exolytic fashion, cf. EC 4.2.2.3 | ? | - |
? | |
4.2.2.11 | sodium alginate | Zobellia galactanivorans DSM 12802 | AlyA1PL7 is an endolytic guluronate lyase that preferentially cleaves guluronate stretches | ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
4.2.2.3 | Zobellia galactanivorans | G0L2Y1 | - |
- |
4.2.2.3 | Zobellia galactanivorans DSM 12802 | G0L2Y1 | - |
- |
4.2.2.11 | Zobellia galactanivorans | G0KZG8 | - |
- |
4.2.2.11 | Zobellia galactanivorans | G0L2Y1 | - |
- |
4.2.2.11 | Zobellia galactanivorans DSM 12802 | G0KZG8 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
4.2.2.3 | recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration to homogeneity | Zobellia galactanivorans |
4.2.2.11 | recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration to homogeneity | Zobellia galactanivorans |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
4.2.2.11 | [alpha-1,4-L-guluronate]n = alpha-L-guluronate + 4-deoxy-alpha-L-erythro-hex-4-enuronosyl-[alpha-1,4-L-guluronate]n-2 | AlyA1PL7 alginate lyase acts via the beta-elimination mechanism, producing the 4,5-unsaturated 4-deoxy-L-erythro-hex-4-enepyranosyluronate (DELTA) on the nonreducing end, catalytic mechanism of G-specific alginate lyases, overview | Zobellia galactanivorans | |
4.2.2.11 | [alpha-1,4-L-guluronate]n = alpha-L-guluronate + 4-deoxy-alpha-L-erythro-hex-4-enuronosyl-[alpha-1,4-L-guluronate]n-2 | AlyA1PL7 alginate lyase acts via the beta-elimination mechanism, producing the 4,5-unsaturated 4-deoxy-L-erythro-hex-4-enepyranosyluronate (DELTA) on the nonreducing end, catalytic mechanism of guluronate-specific alginate lyases, overview | Zobellia galactanivorans |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
4.2.2.3 | additional information | the enzyme has a broad substrate tolerance and can cleave mannuronate-mannuronate, mannuronate-guluronate, and guluronate-guluronate linkages at the nonreducing end. The activity is depending on the block structure | Zobellia galactanivorans | ? | - |
? | |
4.2.2.3 | additional information | the enzyme has a broad substrate tolerance and can cleave mannuronate-mannuronate, mannuronate-guluronate, and guluronate-guluronate linkages at the nonreducing end. The activity is depending on the block structure | Zobellia galactanivorans DSM 12802 | ? | - |
? | |
4.2.2.3 | poly(beta-D-manuronate) | - |
Zobellia galactanivorans | ? | - |
? | |
4.2.2.3 | poly(beta-D-manuronate) | - |
Zobellia galactanivorans DSM 12802 | ? | - |
? | |
4.2.2.3 | sodium alginate | AlyA5 cleaves unsaturated units, alpha-L-guluronate or beta-D-manuronate residues, at the nonreducing end of oligo-alginates in an exolytic fashion, cf. EC 4.2.2.11 | Zobellia galactanivorans | ? | - |
? | |
4.2.2.3 | sodium alginate | LC mass and NMR spectrometris product analysis | Zobellia galactanivorans | ? | - |
? | |
4.2.2.3 | sodium alginate | AlyA5 cleaves unsaturated units, alpha-L-guluronate or beta-D-manuronate residues, at the nonreducing end of oligo-alginates in an exolytic fashion, cf. EC 4.2.2.11 | Zobellia galactanivorans DSM 12802 | ? | - |
? | |
4.2.2.3 | sodium alginate | LC mass and NMR spectrometris product analysis | Zobellia galactanivorans DSM 12802 | ? | - |
? | |
4.2.2.11 | additional information | alginate pentasaccharides that can be cleaved by AlyA1PL7 are GGGGG, GGMGG, GGGMG, and GGMMG | Zobellia galactanivorans | ? | - |
? | |
4.2.2.11 | additional information | the enzyme has a broad substrate tolerance and can cleave M-M, M-G, and G-G linkages at the nonreducing end. The activity is depending on the block structure | Zobellia galactanivorans | ? | - |
? | |
4.2.2.11 | additional information | alginate pentasaccharides that can be cleaved by AlyA1PL7 are GGGGG, GGMGG, GGGMG, and GGMMG | Zobellia galactanivorans DSM 12802 | ? | - |
? | |
4.2.2.11 | additional information | the enzyme has a broad substrate tolerance and can cleave M-M, M-G, and G-G linkages at the nonreducing end. The activity is depending on the block structure | Zobellia galactanivorans DSM 12802 | ? | - |
? | |
4.2.2.11 | poly(alpha-L-guluronate) | - |
Zobellia galactanivorans | ? | - |
? | |
4.2.2.11 | poly(alpha-L-guluronate) | unmodified substrate poly-G blocks and substrate that with its reducing end being reduced using sodium borohydride prior to the digestion | Zobellia galactanivorans | ? | - |
? | |
4.2.2.11 | poly(alpha-L-guluronate) | - |
Zobellia galactanivorans DSM 12802 | ? | - |
? | |
4.2.2.11 | sodium alginate | AlyA1PL7 is an endolytic guluronate lyase that preferentially cleaves guluronate stretches | Zobellia galactanivorans | ? | - |
? | |
4.2.2.11 | sodium alginate | AlyA5 cleaves unsaturated units, alpha-L-guluronate or beta-D-manuronate residues, at the nonreducing end of oligo-alginates in an exolytic fashion, cf. EC 4.2.2.3 | Zobellia galactanivorans | ? | - |
? | |
4.2.2.11 | sodium alginate | AlyA1PL7 is an endolytic guluronate lyase that preferentially cleaves guluronate stretches, no activity with poly-(mannuronate-guluronate) and poly-(mannuronate-mannuronate), minimal recognition pattern of AlyA1PL7, overview | Zobellia galactanivorans | ? | mainly trisaccharide and tetrasaccharide oligomers of alginate with a total content of 41% and 36%, respectively, around 19% disaccharide and only a small amount of pentamers and hexamers, LC mass and NMR spectrometric product analysis | ? | |
4.2.2.11 | sodium alginate | LC mass and NMR spectrometris product analysis | Zobellia galactanivorans | ? | - |
? | |
4.2.2.11 | sodium alginate | AlyA1PL7 is an endolytic guluronate lyase that preferentially cleaves guluronate stretches | Zobellia galactanivorans DSM 12802 | ? | - |
? | |
4.2.2.11 | sodium alginate | AlyA1PL7 is an endolytic guluronate lyase that preferentially cleaves guluronate stretches, no activity with poly-(mannuronate-guluronate) and poly-(mannuronate-mannuronate), minimal recognition pattern of AlyA1PL7, overview | Zobellia galactanivorans DSM 12802 | ? | mainly trisaccharide and tetrasaccharide oligomers of alginate with a total content of 41% and 36%, respectively, around 19% disaccharide and only a small amount of pentamers and hexamers, LC mass and NMR spectrometric product analysis | ? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
4.2.2.3 | monomer or dimer | x * 42000, recombinant His-tagged enzyme, SDS-PAGE | Zobellia galactanivorans |
4.2.2.11 | ? | 2 * 265000-29000, recombinant His-tagged enzyme, SDS-PAGE | Zobellia galactanivorans |
4.2.2.11 | monomer or dimer | x * 42000, recombinant His-tagged enzyme, SDS-PAGE | Zobellia galactanivorans |
4.2.2.11 | More | overall structure of AlyA1PL7 is a PL7 beta-sandwich jelly roll-fold, formed by two anti-parallel beta-sheets stacked against each other. The outer convex sheet is composed of five beta-strands, and the inner concave sheet is composed of seven beta-strands, forming a groove that harbors the catalytic active site | Zobellia galactanivorans |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
4.2.2.3 | AlyA5 | - |
Zobellia galactanivorans |
4.2.2.11 | AlyA1PL7 | - |
Zobellia galactanivorans |
4.2.2.11 | AlyA5 | - |
Zobellia galactanivorans |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
4.2.2.3 | 30 | - |
assay at | Zobellia galactanivorans |
4.2.2.11 | 30 | - |
- |
Zobellia galactanivorans |
4.2.2.11 | 30 | - |
assay at | Zobellia galactanivorans |
EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|---|
4.2.2.3 | 10 | 45 | temperature profile, overview | Zobellia galactanivorans |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.2.2.11 | 12.66 | - |
sodium alginate | with L-guluronate content of 66.7%, pH 7.5, 30°C, recombinant enzyme | Zobellia galactanivorans | |
4.2.2.11 | 17.89 | - |
sodium alginate | with L-guluronate content of 52.6%, pH 7.5, 30°C, recombinant enzyme | Zobellia galactanivorans | |
4.2.2.11 | 19.51 | - |
sodium alginate | with L-guluronate content of 33.3%, pH 7.5, 30°C, recombinant enzyme | Zobellia galactanivorans |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
4.2.2.3 | 7 | - |
- |
Zobellia galactanivorans |
4.2.2.11 | 7 | - |
- |
Zobellia galactanivorans |
4.2.2.11 | 7.5 | - |
assay at | Zobellia galactanivorans |
EC Number | pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|---|
4.2.2.3 | additional information | - |
pH profile, narrow activity range, overview | Zobellia galactanivorans |
4.2.2.11 | additional information | - |
pH profile, narrow activity range, overview | Zobellia galactanivorans |
4.2.2.11 | 4 | 8.5 | no activity outside this range | Zobellia galactanivorans |
EC Number | General Information | Comment | Organism |
---|---|---|---|
4.2.2.3 | evolution | AlyA5 belongs to the PL7 family, subfamily 5, the genome of Zobellia galactanivorans contains seven putative alginate lyase genes, five of which are localized within two clusters comprising additional carbohydrate-related genes, phylogenetic analysis. Despite a common jelly roll-fold, these striking differences of the mode of action are explained by a distinct active site topology, an open cleft in AlyA1PL7, whereas AlyA5 displays a pocket topology due to the presence of additional loops partially obstructing the catalytic groove. In contrast to PL7 alginate lyases from terrestrial bacteria, both enzymes proceed according to a calcium-dependent mechanism | Zobellia galactanivorans |
4.2.2.3 | additional information | secondary structure comparison of AlyA5 and AlyA1PL7 | Zobellia galactanivorans |
4.2.2.11 | evolution | AlyA1PL7 is an endolytic guluronate lyase and belongs to the PL7 family, subfamily 1, the genome of Zobellia galactanivorans contains seven putative alginate lyase genes, five of which are localized within two clusters comprising additional carbohydrate-related genes, phylogenetic analysis. Despite a common jelly roll-fold, these striking differences of the mode of action are explained by a distinct active site topology, an open cleft in AlyA1PL7, whereas AlyA5 displays a pocket topology due to the presence of additional loops partially obstructing the catalytic groove. In contrast to PL7 alginate lyases from terrestrial bacteria, both enzymes proceed according to a calcium-dependent mechanism | Zobellia galactanivorans |
4.2.2.11 | evolution | AlyA5 belongs to the PL7 family, subfamily 5, the genome of Zobellia galactanivorans contains seven putative alginate lyase genes, five of which are localized within two clusters comprising additional carbohydrate-related genes, phylogenetic analysis. Despite a common jelly roll-fold, these striking differences of the mode of action are explained by a distinct active site topology, an open cleft in AlyA1PL7, whereas AlyA5 displays a pocket topology due to the presence of additional loops partially obstructing the catalytic groove. In contrast to PL7 alginate lyases from terrestrial bacteria, both enzymes proceed according to a calcium-dependent mechanism | Zobellia galactanivorans |
4.2.2.11 | additional information | secondary structure comparison of AlyA5 and AlyA1PL7 | Zobellia galactanivorans |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.2.2.11 | 3.16 | - |
sodium alginate | with L-guluronate content of 33.3%, pH 7.5, 30°C, recombinant enzyme | Zobellia galactanivorans | |
4.2.2.11 | 5.79 | - |
sodium alginate | with L-guluronate content of 52.6%, pH 7.5, 30°C, recombinant enzyme | Zobellia galactanivorans | |
4.2.2.11 | 7.22 | - |
sodium alginate | with L-guluronate content of 66.7%, pH 7.5, 30°C, recombinant enzyme | Zobellia galactanivorans |