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Literature summary for 2.7.1.17 extracted from

  • Akinterinwa, O.; Cirino, P.C.
    Heterologous expression of d-xylulokinase from Pichia stipitis enables high levels of xylitol production by engineered Escherichia coli growing on xylose (2009), Metab. Eng., 11, 48-55.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli strain PC07 with deleted xylB-gene and introduced plasmid with Escherichia coli xylB gene Escherichia coli

Protein Variants

Protein Variants Comment Organism
additional information Escherichia coli xylulokinase gene xylB is PCR-amplified from Escherichia coli genomic DNA, genes of interest are cloned downstream of a tac promoter and upstream of a transcription termination sequence into the medium copy vector pLOI3809, transforming plasmid into a DELTAxylB mutant E. coli strain (PC07) Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + D-xylulose Escherichia coli
-
ADP + D-xylulose 5-phosphate
-
r

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
DELTAxylB mutant strain (PC07), xylB encodes XK in Escherichia coli
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
3.8 U/mg protein (native substrate 5 mM xylulose), 0.76 U/mg protein (substrate 75 mM xylitol), activities measured from cell lysates by monitoring NADH depletion, using an assay in which pentulokinase activity is coupled to pyruvate kinase and lactate dehydrogenase activities, reaction mixture (final concentrations indicated): 71 mM Tris–HCl pH 7.5, 7.1 mM MgCl2, 1 mM EDTA, 50 mM KCl, 7.1 mM KF, 5 mM KCN, 1.4 mM ATP, 1 mM phosphoenolpyruvic acid, 0.3 mM NADH, 0.7 U/ml pyruvate kinase, 1 U/ml lactate dehydrogenase, and 5 mM D-xylulose or 75 mM xylitol Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + D-xylitol analyzed with 31P NMR spectrum Escherichia coli ADP + D-xylitol-1-phosphate
-
?
ATP + D-xylitol analyzed with 31P NMR spectrum Escherichia coli ADP + D-xylitol-5-phosphate
-
?
ATP + D-xylulose
-
Escherichia coli ADP + D-xylulose 5-phosphate
-
r

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
Escherichia coli