Figure 3. Decrease in the amount of lipid intermediates and peptidoglycan synthesis in mvaA mutants.

(a) Illustration of the synthesis of peptideglycan from FPP. GlcNAc: β-1-4-N-acetylglucosamine, MurNAc: N-acetylmuramic acid. (b) Overnight cultures of RN4220 (wild-type), TSJY1, TSJY2, and TSJY3 were diluted 1000-fold, and further incubated at 30 °C or 43 °C for 5 h. Bacterial cells were disrupted and separated into a supernatant fraction and wall-membrane particulate fraction. The products of the in vitro reaction with [¹⁴C]glycine were separated by thin-layer chromatography as described in the Experimental Procedures. (c) Overnight cultures of RN4220 (wild-type), TSJY1, TSJY2, and TSJY3 were diluted 1000-fold, and further incubated at 30 °C or 43 °C. Cultures (OD₆₀₀ = 0.1–0.5) were adjusted to OD₆₀₀ = 0.5 in cell wall synthesis medium (CWSM) containing chloramphenicol (100 μg/ml), followed by incubation with [¹⁴C] N-acetylglucosamine at 30 °C or 43 °C for 30 min. Peptidoglycan synthesis was measured by [¹⁴C] N-acetylglucosamine incorporation. *P < 0.05. (d) Overnight cultures of RN4220 (wild-type), TSJY1, TSJY2, and TSJY3 were diluted 1000-fold, and further incubated with or without mevalonate (500 μM) at 30 °C or 43 °C. Cultures (OD₆₀₀ = 0.1–0.5) were adjusted to OD₆₀₀ = 0.5 in CWSM containing chloramphenicol (100 μg/ml), followed by incubation with [¹⁴C] N-acetylglucosamine with or without mevalonate (5 mM) in CWSM containing chloramphenicol (100 μg/ml) at 43 °C for 30 min. Peptidoglycan synthesis was measured by [¹⁴C] N-acetylglucosamine incorporation. Mev: mevalonate. *P < 0.05