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Archaea. 2012; 2012: 438931.
Published online May 9, 2012. doi:  10.1155/2012/438931
PMCID: PMC3357500
Archaeal Phospholipid Biosynthetic Pathway Reconstructed in Escherichia coli
Takeru Yokoi, Keisuke Isobe, Tohru Yoshimura, and Hisashi Hemmi *
Department of Applied Molecular Bioscience, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 460-8601, Japan
*Hisashi Hemmi: hhemmi/at/agr.nagoya-u.ac.jp
Academic Editor: Yosuke Koga
Received December 19, 2011; Accepted February 23, 2012.
Abstract
A part of the biosynthetic pathway of archaeal membrane lipids, comprised of 4 archaeal enzymes, was reconstructed in the cells of Escherichia 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 vitro radioassay 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 in E. coli. Introduction of the 4 genes into E. coli resulted 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 of E. coli.
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