State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
Biotechnol Lett. 2011 May;33(5):1013-9. doi: 10.1007/s10529-011-0521-z. Epub 2011 Jan 19.
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.
脂质 A 是大多数革兰氏阴性菌外膜的主要组成部分。单磷酸脂质 A 在 1 位不含磷酸基团，可用作佐剂。我们通过将 lpxE 基因整合到大肠杆菌 W3110 的染色体中，构建了一个大肠杆菌突变体 CW001。该基因 lpxE 编码一种酶 LpxE，它可以去除脂质 A 的 1-磷酸基团。通过薄层层析和电喷雾电离质谱分析，CW001 主要在体内产生 1-去磷酸化的脂质 A。这项研究不仅对脂质 A 佐剂的开发具有重要意义，而且为将异源基因整合到大肠杆菌染色体中提供了一种新方法。
