Construction of an Escherichia coli mutant producing monophosphoryl lipid A.
Lipid A is a major component in the outer membrane of most gram-negative bacteria. Monophosphoryl lipid A contains no phosphate group at 1-position and can be used as an adjuvant. We constructed an Escherichia coli mutant CW001 by integrating a gene lpxE into the chromosome of E. coli W3110. The gene lpxE encodes an enzyme LpxE which removes the 1-phosphate group of lipid A. CW001 predominantly produces 1-dephosphorylated lipid A in vivo, as adjudged by thin layer chromatography and electro-spray ionization mass spectrometry. This study not only is important for the development of lipid A adjuvants but also provides a novel method for integration of heterologous genes into the chromosome of E. coli.
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
Jiuzhou
J
Chen
Guanjun
G
Tao
Xiaoyuan
X
Wang
D002855:Chromatography, Thin Layer; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008050:Lipid A / Q000031:analogs & derivatives* / Q000096:biosynthesis; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D012475:Salmonella / Q000201:enzymology / Q000235:genetics; D021241:Spectrometry, Mass, Electrospray Ionization; D014169:Transformation, Bacterial
D008050:Lipid A; D010744:Phosphoric Monoester Hydrolases; C048436:monophosphoryl lipid A