Isoprenoid synthesis in Escherichia coli. Separation and partial purification of four enzymes involved in the synthesis.
Isopentenyl pyrophosphate (IPP) isomerase, farnesyl pyrophosphate (FPP) synthetase, octaprenyl pyrophosphate (OPP) synthetase and undecaprenyl pyrophosphate (UPP) synthetase were partially purified from Escherichia coli by DEAE-Toyopearl chromatography. FPP synthetase catalyzed the condensation of IPP with dimethylallyl pyrophosphate (DPP) as well as with geranyl pyrophosphate (GPP) to yield FPP as final product. OPP synthetase and UPP synthetase catalyzed the condensation of IPP with FPP to yield OPP and cis,trans-polyprenyl pyrophosphates (the C45-, C50, and C55-compound), respectively. Neither DPP nor GPP acted as a priming substrate for either enzyme. These four enzymes required Mg2+ or Mn2+ for their activities. UPP synthetase required also Triton X-100 for its activity. The addition of Triton X-100 enhanced OPP synthetase, but it did not affect IPP isomerase and FPP synthetase. It seems possible that the combination of the four enzymes ensures the in vivo synthesis of long-chain isoprenoids in E. coli.
D019883:Alkyl and Aryl Transferases; D019748:Carbon-Carbon Double Bond Isomerases; D002384:Catalysis; D002852:Chromatography, Ion Exchange; D002855:Chromatography, Thin Layer; D004122:Dimethylallyltranstransferase / Q000302:isolation & purification; D004789:Enzyme Activation / Q000187:drug effects; D004926:Escherichia coli / Q000201:enzymology*; D045782:Hemiterpenes; D006863:Hydrogen-Ion Concentration; D007535:Isomerases / Q000302:isolation & purification; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D011106:Polyisoprenyl Phosphates / Q000096:biosynthesis*; D013379:Substrate Specificity; D014166:Transferases / Q000302:isolation & purification
D045782:Hemiterpenes; D008670:Metals; D011106:Polyisoprenyl Phosphates; D011092:Polyethylene Glycols; D017830:Octoxynol; D014166:Transferases; D019883:Alkyl and Aryl Transferases; C048987:octaprenyl pyrophosphate synthetase; D004122:Dimethylallyltranstransferase; C023266:undecaprenyl pyrophosphate synthetase; D007535:Isomerases; D019748:Carbon-Carbon Double Bond Isomerases; C021453:isopentenyldiphosphate delta-isomerase