Activation of a cryptic gene encoding a kinase for L-xylulose opens a new pathway for the utilization of L-lyxose by Escherichia coli.
A silent gene encoding a kinase that specifically phosphorylates L-xylulose was activated and rendered constitutive in mutant cells of Escherichia coli. L-Xylulose kinase was purified to homogeneity and found to be a dimer of two subunits of 55 kDa, highly specific for L-xylulose with a Km of 0.8 mM, a Vmax of 33 mumol/min/mg, and an optimum pH of 8.4. Physical (thin layer chromatography) and spectroscopic (nuclear magnetic resonance and optical rotation) characterization of the product of L-xylulose kinase indicated that the enzyme phosphorylated the sugar at position 5. The gene encoding L-xylulose kinase was mapped in the 80.2 min region of the chromosome by conjugation and transduction. Cloning and comparison of the restriction map with the Kohara map (Kohara, Y., Akiyame, K., and Isono, K. (1987) Cell 50, 495-501) located the gene between positions 3963 and 3965 kilobases. The molecular and functional features of L-xylulose kinase together with the location of the corresponding gene indicate that this enzyme did not derive from mutation of any other known kinase. The new kinase opens a route for the utilization of L-lyxose through the action of rhamnose permease, rhamnose isomerase, and the phosphorylation of the L-xylulose formed to L-xylulose 5-phosphate, which is then introduced into the pentose phosphate pathway for subsequent metabolism.
Department of Biochemistry, School of Pharmacy, University of Barcelona, Spain.
J C
JC
Sánchez
R
R
Gímenez
A
A
Schneider
W D
WD
Fessner
L
L
Baldomà
J
J
Aguilar
J
J
Badía
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism*; D015870:Gene Expression; D009154:Mutation; D010428:Pentosephosphates / Q000378:metabolism; D010429:Pentoses / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping; D012210:Rhamnose / Q000378:metabolism; D013379:Substrate Specificity; D014996:Xylulose / Q000378:metabolism*
D010428:Pentosephosphates; D010429:Pentoses; D014996:Xylulose; C031625:xylulose-5-phosphate; C043333:lyxose; D017853:Phosphotransferases (Alcohol Group Acceptor); C032700:xylulokinase; D012210:Rhamnose