Archaeal Phospholipid Biosynthetic Pathway Reconstructed inEscherichia coli

[摘要] A part of the biosynthetic pathway of archaeal membrane lipids, comprised of 4 archaeal enzymes, was reconstructed in the cells ofEscherichia coli. The genes of the enzymes were cloned from a mesophilic methanogen,Methanosarcina acetivorans, and the activity of each enzyme was confirmed using recombinant proteins.In vitroradioassay showed that the 4 enzymes are sufficient to synthesize an intermediate of archaeal membrane lipid biosynthesis, that is,2,3-di-O-geranylgeranyl-sn-glycerol-1-phosphate, from precursors that can be produced endogenously inE. coli. Introduction of the 4 genes intoE. coliresulted in the production of archaeal-type lipids. Detailed liquid chromatography/electron spray ionization-mass spectrometry analyses showed that they are metabolites from the expected intermediate, that is, 2,3-di-O-geranylgeranyl-sn-glycerol and 2,3-di-O-geranylgeranyl-sn-glycerol-1-phosphoglycerol. The metabolic processes, that is, dephosphorylation and glycerol modification, are likely catalyzed by endogenous enzymes ofE. coli.

[学科分类] 微生物学和免疫学