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Literature summary extracted from

  • Qi, Z.; Zhu, Z.; Wang, J.W.; Li, S.; Guo, Q.; Xu, P.; Lu, F.; Qin, H.M.
    Biochemical analysis and the preliminary crystallographic characterization of D-tagatose 3-epimerase from Rhodobacter sphaeroides (2017), Microb. Cell Fact., 16, 193 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.1.3.31 gene RSP_3671, sequence comparisons and phylogenetic tree, recombinant expression in Escherichia coli strain BL21(DE3) Cereibacter sphaeroides

Protein Variants

EC Number Protein Variants Comment Organism
5.1.3.31 R118W site-directed mutagenesis, the unique hydrogen bond between Arg118 and O4 of D-fructose is broken when Arg118 is mutated to Trp, the mutation improves the substrate recognition and activity of the enzyme. The mutant enzyme RsDTE_R118W shows decreased catalytic activity compared to the wild-type enzyme toward D-fructose, the kcat/Km for D-tagatose is about twofold higher than for D-psicose. Mutant R118W shows 1.5fold higher catalytic efficiency toward D-tagatose than the wild-type Cereibacter sphaeroides

Inhibitors

EC Number Inhibitors Comment Organism Structure
5.1.3.31 Ca2+
-
 Cereibacter sphaeroides
5.1.3.31 Cu2+ high inhibition at 0.1 mM Cereibacter sphaeroides
5.1.3.31 EDTA
-
 Cereibacter sphaeroides
5.1.3.31 Fe2+ high inhibition at 0.1 mM Cereibacter sphaeroides
5.1.3.31 Fe3+
-
 Cereibacter sphaeroides
5.1.3.31 Ni2+ almost complete inhibition at 0.1 mM Cereibacter sphaeroides
5.1.3.31 Zn2+
-
 Cereibacter sphaeroides

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.1.3.31 additional information
-
 additional information Michaelis-Menten kinetics Cereibacter sphaeroides
5.1.3.31 0.98
-
 D-tagatose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 78
-
 D-fructose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 137.9
-
 D-tagatose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 148.1
-
 D-sorbose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 162.1
-
 D-fructose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 192.9
-
 D-psicose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 215.2
-
 D-psicose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 359.4
-
 D-sorbose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
5.1.3.31 Co2+ dependent on, activates at 0.1 mM, metalloenzyme Cereibacter sphaeroides
5.1.3.31 Mn2+ dependent on, activates at 0.1 mM, metalloenzyme, Mn2+ is the most effective activating divalent cation Cereibacter sphaeroides
5.1.3.31 additional information no effect on enzyme activity by Mg2+ at .1 mM Cereibacter sphaeroides

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.1.3.31 D-fructose Cereibacter sphaeroides
-
 D-psicose
-
 r
5.1.3.31 D-fructose Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158
-
 D-psicose
-
 r
5.1.3.31 D-tagatose Cereibacter sphaeroides
-
 D-sorbose
-
 r

Organism

EC Number Organism UniProt Comment Textmining
5.1.3.31 Cereibacter sphaeroides Q3IW04
-
-
5.1.3.31 Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158 Q3IW04
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.1.3.31 D-fructose
-
 Cereibacter sphaeroides D-psicose
-
 r
5.1.3.31 D-fructose best substrate for the recombinant wild-type enzyme Cereibacter sphaeroides D-psicose
-
 r
5.1.3.31 D-fructose
-
 Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158 D-psicose
-
 r
5.1.3.31 D-fructose best substrate for the recombinant wild-type enzyme Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158 D-psicose
-
 r
5.1.3.31 D-psicose
-
 Cereibacter sphaeroides D-fructose
-
 r
5.1.3.31 D-psicose
-
 Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158 D-fructose
-
 r
5.1.3.31 D-sorbose
-
 Cereibacter sphaeroides D-tagatose
-
 r
5.1.3.31 D-sorbose
-
 Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158 D-tagatose
-
 r
5.1.3.31 D-tagatose
-
 Cereibacter sphaeroides D-sorbose
-
 r
5.1.3.31 D-tagatose best substrate for the recombinant enzyme mutant R118W Cereibacter sphaeroides D-sorbose
-
 r
5.1.3.31 additional information RsDTE wild-type shows lower Michaelis-Menten constant (Km), lower turnover number (kcat), but higher catalytic efficiency (kcat/Km) values for D-fructose than for D-psicose. The kcat/Km for D-fructose is 5.5fold higher than for D-psicose, indicating that enzyme RsDTE highly catalyzes D-fructose, although it is a D-tagatose 3-epimerase Cereibacter sphaeroides ?
-
 ?
5.1.3.31 additional information RsDTE wild-type shows lower Michaelis-Menten constant (Km), lower turnover number (kcat), but higher catalytic efficiency (kcat/Km) values for D-fructose than for D-psicose. The kcat/Km for D-fructose is 5.5fold higher than for D-psicose, indicating that enzyme RsDTE highly catalyzes D-fructose, although it is a D-tagatose 3-epimerase Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
5.1.3.31 RsDTE
-
 Cereibacter sphaeroides
5.1.3.31 RSP_3671
-
 Cereibacter sphaeroides

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.1.3.31 35
-
-
 Cereibacter sphaeroides

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
5.1.3.31 20 70 half-maximal activity at 20°C, near inactivation at 70°C Cereibacter sphaeroides

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.1.3.31 8.3
-
 D-sorbose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 15.36
-
 D-sorbose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 20.47
-
 D-psicose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 26.04
-
 D-psicose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 41.05
-
 D-fructose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 42.6
-
 D-fructose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 48.31
-
 D-tagatose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 51.25
-
 D-tagatose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.1.3.31 8.5
-
-
 Cereibacter sphaeroides

pH Range

EC Number pH Minimum pH Maximum Comment Organism
5.1.3.31 5.5 11 activity range Cereibacter sphaeroides

General Information

EC Number General Information Comment Organism
5.1.3.31 evolution the enzyme belongs to the D-tagatose 3-epimerase (D-TE) family Cereibacter sphaeroides
5.1.3.31 additional information the O1 of substrate D-fructose forms hydrogen bonds with His192 and Glu162 to help the correct metal coordination of the substrate. The O2 forms hydrogen bonds with His192 and Arg221. Glu156 forms hydrogen bonds with O3, and Glu250 directs its OE2 atom to a hydrogen atom attached to C3. Because D-fructose has the same configurations of C1, C2 and C3 as D-tagatose, the interactions between D-fructose at the 1-, 2- and 3-positions and the enzyme are very similar to those in other DTE/DPE family enzymes. Residue R118 forms a hydrogen bond with O4 of D-fructose and may regulate the substrate specificity. The strengthened hydrophobic interaction may attribute to the recognition of D-tagatose, D-psicose, and D-sorbose. Enzyme homology modeling and structure comparisons, overview Cereibacter sphaeroides
5.1.3.31 physiological function the purified D-tagatose 3-epimerase from Rhodobacter sphaeroides catalyzes the epimerization of D-fructose to D-psicose at the C3 position Cereibacter sphaeroides

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.1.3.31 0.02
-
 D-sorbose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 0.1
-
 D-sorbose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 0.1
-
 D-psicose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 0.14
-
 D-psicose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 0.25
-
 D-fructose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 0.35
-
 D-tagatose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 0.52
-
 D-tagatose recombinant mutant R118W, pH 8.5, 35°C Cereibacter sphaeroides
5.1.3.31 0.55
-
 D-fructose recombinant wild-type enzyme, pH 8.5, 35°C Cereibacter sphaeroides