Cloned (Comment) | Organism |
---|---|
gene flkin, DNA and amino acid sequence determination and analysis, genetic organization of alginate-assimilating enzyme genes in the genome of Flavobacterium sp. strain UMI-01, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Flavobacterium sp. UMI-01 |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of in vitro alginate-metabolizing system using recombinant enzymes. Alginate is allowed to react with recFlAlyA, recFlAlyB, recFlAlex, recFlRed, recFlKin, and recFlAld in various combinations at 25°C | Flavobacterium sp. UMI-01 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Flavobacterium sp. UMI-01 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 2-dehydro-3-deoxy-D-gluconate | Flavobacterium sp. UMI-01 | - |
ADP + 2-dehydro-3-deoxy-6-phospho-D-gluconate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Flavobacterium sp. UMI-01 | A0A0B6VMB0 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Flavobacterium sp. UMI-01 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.72 | - |
purified recobinant enzyme, pH 7.4, 30°C | Flavobacterium sp. UMI-01 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 2-dehydro-3-deoxy-D-gluconate | - |
Flavobacterium sp. UMI-01 | ADP + 2-dehydro-3-deoxy-6-phospho-D-gluconate | - |
? | |
additional information | thin-layer chromatography (TLC) and mass spectrometric analyses of reaction products of recombinant FlKin | Flavobacterium sp. UMI-01 | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
? | x * 39391, sequence calculation, x * 39000, recombinant enzyme, SDS-PAGE | Flavobacterium sp. UMI-01 |
Synonyms | Comment | Organism |
---|---|---|
2-keto-3-deoxy-D-gluconate kinase | - |
Flavobacterium sp. UMI-01 |
FlKin | - |
Flavobacterium sp. UMI-01 |
KDG kinase | - |
Flavobacterium sp. UMI-01 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
recombinant enzyme | Flavobacterium sp. UMI-01 |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
purified recombinant enzyme, pH 7.0, 30 min, stable | Flavobacterium sp. UMI-01 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
recombinant enzyme | Flavobacterium sp. UMI-01 |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | - |
purified recombinant enzyme, 40°C, 30 min, stable | Flavobacterium sp. UMI-01 |
General Information | Comment | Organism |
---|---|---|
evolution | 2-keto-3-deoxy-D-gluconate (KDG) kinase and 2-keto-3-deoxy-6-phosphogluconate (KDPG) aldolase genes, i.e., flkin and flald, are encoded in the assimilating gene cluster in the genome of Flavobacterium sp. strain UMI-01, a member of Bacteroidetes. The amino acid sequences deduced from flkin and flald show low identities with those of corresponding enzymes of Saccharophagus degradans strain 2-40T, a member of Proteobacteria | Flavobacterium sp. UMI-01 |
metabolism | the enzyme is active in alginate degradation. Unsaturated monosaccharide, the end product of alginate lyases, is spontaneously and/or enzymatically converted to an open chain form, DEH, and further converted to 2-keto-3-deoxy-D-gluconate (KDG) by the NAD(P)H-dependent DEH reductase. The KDG is phosphorylated to KDPG by KDG kinase and then split to pyruvate and glyceraldehyde-3-phosphate (GAP) by KDPG aldolase. The alginate-derived pyruvate and GAP are finally metabolized by Kreb's cycle. Alginate-metabolic system of Flavobacterium sp. strain UMI-01, overview | Flavobacterium sp. UMI-01 |
physiological function | the enzyme is active in alginate degradation | Flavobacterium sp. UMI-01 |