Method of producing lipidated polypeptides

A method of producing a recombinant lipidated polypeptide in E. coli, the method comprising:

• providing an E. coli host cell adapted to express a recombinant lipidated polypeptide, wherein the E. coli host cell is C41 (DE3), C43 (DE3), DK8 (DE3)S, or C2014 (DE3) and has a nucleic acid sequence encoding the recombinant lipidated polypeptide, which contains a bacterial lipoprotein signal peptide;
• and
• culturing the E. coli host cell in a M9 medium under conditions that allow expression of the polypeptide in lipidated form, wherein the ratio of immature to mature lipoprotein (I/M ratio) of the recombinant lipidated polypeptide thus expressed is 0.01 or lower, the M9 medium lacking an organic nitrogen source and a ferrous source, containing 100 mM or less of phosphate and glucose as the sole carbon source, and having a pH of 7.0-8.5.