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Literature summary for 2.4.1.14 extracted from

  • Sawitri, W.D.; Narita, H.; Ishizaka-Ikeda, E.; Sugiharto, B.; Hase, T.; Nakagawa, A.
    Purification and characterization of recombinant sugarcane sucrose phosphate synthase expressed in E. coli and insect Sf9 cells: an importance of the N-terminal domain for an allosteric regulatory property (2016), J. Biochem., 159, 599-607.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
glucose 6-phosphate allosteric activation of the wild-type enzyme, N-terminally truncated enzymes are not activated Saccharum officinarum

Cloned(Commentary)

Cloned (Comment) Organism
gene SoSPS1, recombinant expression of N-terminaly His6-tagged wild-type enzyme and truncated mutant enzymes lacking up to 171 residues of the N-terminal region in Escherichia coli strain BL21(DE3), recombinant expression of only the full-length wild-type enzyme with a His8-myc-His8 tag, a TEV cleavage site, and EGFP fusion in Spodoptera frugiperda Sf9 cells, the latter via baculovirus transfection Saccharum officinarum

Protein Variants

Protein Variants Comment Organism
additional information construction of N-terminally truncated enzymeslacking up to 171 amino acid residues in the 20 kDa N-terminal region, the mutants cannot be activated Saccharum officinarum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information allosteric regulation by glucose 6-phosphate Saccharum officinarum

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Saccharum officinarum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-glucose + D-fructose 6-phosphate Saccharum officinarum
-
UDP + sucrose 6F-phosphate
-
?

Organism

Organism UniProt Comment Textmining
Saccharum officinarum P93782 gene SoSPS1
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) and His8-myc-His8 tagged EGFP-enzyme from insect Sf9 cells by anion exchange and nickel affinity chromatography, followed by gel filtration. Purification of the full-length enzyme without His-tag by ion exchange and hydrophobic interaction chromatography, and gel filtration Saccharum officinarum

Source Tissue

Source Tissue Comment Organism Textmining
leaf
-
Saccharum officinarum
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-glucose + D-fructose 6-phosphate
-
Saccharum officinarum UDP + sucrose 6F-phosphate
-
?

Subunits

Subunits Comment Organism
More two forms of recombinant enzyme, form A of 100 kDa and form B of 120 kDa, in Escherichia coli. Form B might be enzymatically more active than form A Saccharum officinarum

Synonyms

Synonyms Comment Organism
SPS
-
Saccharum officinarum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25 27 assay at Saccharum officinarum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Saccharum officinarum

General Information

General Information Comment Organism
additional information the N-terminal region of sugarcane sucrose phosphate synthase is not essential for the catalytic reaction itself, but is crucial for the allosteric regulation by glucose 6-phosphate and may function like a suppressor domain for the enzyme activity Saccharum officinarum