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Literature summary for 2.7.1.40 extracted from

  • Hofmann, J.; Heider, C.; Li, W.; Krausze, J.; Roessle, M.; Wilharm, G.
    Recombinant production of Yersinia enterocolitica pyruvate kinase isoenzymes PykA and PykF (2013), Protein Expr. Purif., 88, 243-247.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
D-fructose-1,6-bisphosphate allosteric effector, binds to PykF but not to PykA Yersinia enterocolitica

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of isozymes PykA and PykF in Escherichia coli. Solubility of both PykA and PykF is superior when expression cultures are incubated at 27°C compared to 37°C, so that for large-scale production both proteins are expressed at 27°C after induction for 5 h Yersinia enterocolitica

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant untagged enzyme, synchrotron radiation small-angle X-ray scattering structure determination and analysis Yersinia enterocolitica

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Yersinia enterocolitica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
50500
-
4 * 50500, PykF, SDS-PAGE, circular dichroism spectroscopy, and gel filtration, 4 * 51500, PykA, SDS-PAGE, circular dichroism spectroscopy, and gel filtration Yersinia enterocolitica
51500
-
4 * 50500, PykF, SDS-PAGE, circular dichroism spectroscopy, and gel filtration, 4 * 51500, PykA, SDS-PAGE, circular dichroism spectroscopy, and gel filtration Yersinia enterocolitica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + pyruvate Yersinia enterocolitica
-
ADP + phosphoenolpyruvate
-
?
ATP + pyruvate Yersinia enterocolitica WA-314
-
ADP + phosphoenolpyruvate
-
?

Organism

Organism UniProt Comment Textmining
Yersinia enterocolitica
-
isozymes PykA and PykF encoded by genes pykA and pykF
-
Yersinia enterocolitica WA-314
-
isozymes PykA and PykF encoded by genes pykA and pykF
-

Purification (Commentary)

Purification (Comment) Organism
recombinant untagged isozymes PykA and PykF 2.71fold and 2.1fold, respectively, from Escherichia coli to near homogeneity by anion exchange chromatography and gel filtration Yersinia enterocolitica

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
85
-
purified recombinant PykA, pH 7.5, 30°C Yersinia enterocolitica
108
-
purified recombinant PykF, pH 7.5, 30°C Yersinia enterocolitica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + pyruvate
-
Yersinia enterocolitica ADP + phosphoenolpyruvate
-
?
ATP + pyruvate
-
Yersinia enterocolitica WA-314 ADP + phosphoenolpyruvate
-
?

Subunits

Subunits Comment Organism
tetramer 4 * 50500, PykF, SDS-PAGE, circular dichroism spectroscopy, and gel filtration, 4 * 51500, PykA, SDS-PAGE, circular dichroism spectroscopy, and gel filtration Yersinia enterocolitica

Synonyms

Synonyms Comment Organism
PykA
-
Yersinia enterocolitica
PykF
-
Yersinia enterocolitica

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Yersinia enterocolitica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Yersinia enterocolitica

Cofactor

Cofactor Comment Organism Structure
ATP
-
Yersinia enterocolitica