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Literature summary for 5.5.1.27 extracted from
- Characterization of a novel Agrobacterium tumefaciens galactarolactone cycloisomerase enzyme for direct conversion of D-galactarolactone to 3-deoxy-2-keto-L-threo-hexarate (2012), J. Biol. Chem., 287, 17662-17671.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene gci, DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli, recombinant expression of Strep-tagged enzyme in Escherichia coli strain BL21(DE3) | Agrobacterium tumefaciens |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| EDTA | compete inhibition at 1 mM, addition of M2+ to the apoenzyme results in a complete restoration of the original activity | Agrobacterium tumefaciens |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | no Michaelis-Menten kinetics | Agrobacterium tumefaciens |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | depedent on | Agrobacterium tumefaciens | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 41000 | - |
8 * 41000, SDS-PAGE, 8 * 42600, about, sequence calculation | Agrobacterium tumefaciens |
| 42600 | - |
8 * 41000, SDS-PAGE, 8 * 42600, about, sequence calculation | Agrobacterium tumefaciens |
| 340900 | - |
gel filtration | Agrobacterium tumefaciens |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| D-galactarolactone | Agrobacterium tumefaciens | - |
3-deoxy-2-dehydro-L-threo-hexarate | - |
? | |
| D-galactarolactone | Agrobacterium tumefaciens C58 / ATCC 33970 | - |
3-deoxy-2-dehydro-L-threo-hexarate | - |
? | |
| D-glucarolactone | Agrobacterium tumefaciens | - |
3-deoxy-2-dehydro-L-threo-hexarate | - |
? | |
| D-glucarolactone | Agrobacterium tumefaciens C58 / ATCC 33970 | - |
3-deoxy-2-dehydro-L-threo-hexarate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Agrobacterium tumefaciens | A9CEQ8 | - |
- |
| Agrobacterium tumefaciens C58 / ATCC 33970 | A9CEQ8 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant Strep-tagged enzyme from Escherichia coli strain BL21(DE3) by affinity chromatography and gel filtration, recombinant enzyme from Escherichia coli by ammonium sulfate fractionation, hydrophobic interaction chromatography, gel filtration, and two steps of anion exchange chromatography | Agrobacterium tumefaciens |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| D-galactaro-1,4-lactone = 5-dehydro-4-deoxy-D-glucarate | reaction mechanism, the reaction product of At Gci from D-galactaro-1,4-lactone and D-glucaro-1,4-lactone conversion is in both cases the L-threo form of 3-deoxy-2-keto-hexarate, NMR spectroscopy analysi. The reaction has to be initiated by proton abstraction from C5 followed by leaving of the 4-OH group.The leaving group is the hydroxyl that has formed the lactone ring with the carboxyl group at C1. The proton abstraction takes place next to the carboxylic acid pointing out from the lactone rings | Agrobacterium tumefaciens |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| D-galactarolactone | - |
Agrobacterium tumefaciens | 3-deoxy-2-dehydro-L-threo-hexarate | - |
? | |
| D-galactarolactone | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | 3-deoxy-2-dehydro-L-threo-hexarate | - |
? | |
| D-glucarolactone | - |
Agrobacterium tumefaciens | 3-deoxy-2-dehydro-L-threo-hexarate | - |
? | |
| D-glucarolactone | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | 3-deoxy-2-dehydro-L-threo-hexarate | - |
? | |
| additional information | the enzyme is active both on D-galactarolactone and D-glucarolactone, but does not work on the corresponding linear hexaric acid forms. The reaction product of At Gci from D-galactaro-1,4-lactone and D-glucaro-1,4-lactone conversion is in both cases the L-threo form of linear 3-deoxy-2-keto-hexarate. Coupled assay with Agrobacterium tumefaciens uronate dehydrogenase | Agrobacterium tumefaciens | ? | - |
? | |
| additional information | the enzyme is active both on D-galactarolactone and D-glucarolactone, but does not work on the corresponding linear hexaric acid forms. The reaction product of At Gci from D-galactaro-1,4-lactone and D-glucaro-1,4-lactone conversion is in both cases the L-threo form of linear 3-deoxy-2-keto-hexarate. Coupled assay with Agrobacterium tumefaciens uronate dehydrogenase | Agrobacterium tumefaciens C58 / ATCC 33970 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homooctamer | 8 * 41000, SDS-PAGE, 8 * 42600, about, sequence calculation | Agrobacterium tumefaciens |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| galactarolactone cycloisomerase | - |
Agrobacterium tumefaciens |
| GCI | - |
Agrobacterium tumefaciens |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 22 | - |
assay at | Agrobacterium tumefaciens |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.8 | - |
assay at | Agrobacterium tumefaciens |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to the large mechanistically diverse enolase superfamily, mandelate racemase subgroup | Agrobacterium tumefaciens |
| metabolism | the enzyme is involved in the oxidative pathway for D-galacturonate catabolism, overview | Agrobacterium tumefaciens |
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Release 2023.1 (January 2023)
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