Comparison of quenching and extraction methodologies for metabolome analysis of Lactobacillus plantarum

Table 4 Intracellular concentrations of ATP, NAD⁺ and G-6P in L. plantarum cells that were extracted with different methods.

Extraction methods		ATP (mM)	NAD⁺ (mM)	G-6P (mM)
A	Cold MeOH	0.05 ± 0.01	8.4 ± 0.6	10.2 ± 0.8
	Perchloric acid	1.07 ± 0.19	13.9 ± 1.2	13.2 ± 1.1
	Hot EtOH	1.15 ± 0.07	14.7 ± 0.9	16.4 ± 0.6
	CHCl₃/H₂O	0.49 ± 0.08	13.9 ± 0.4	15.2 ± 0.6
	CHCl₃/MeOH	0.15 ± 0.02	8.6 ± 0.9	11.4 ± 0.3
B	Modified Cold MeOH	1.53 ± 0.15	11.8 ± 0.4	10.5 ± 0.7
	Perchloric acid	1.05 ± 0.01	11.5 ± 2.2	11.0 ± 1.3
	Hot EtOH	1.38 ± 0.34	9.0 ± 0.1	9.4 ± 1.0

Intracellular pools of ATP, NAD⁺ and G-6P in L. plantarum cells that were extracted with five different methods. These amounts and their relative standard deviation are from different extracts. The cells were harvested without quenching, washed, concentrated. Each sample in the top part of the table (A section) contained 9.8 mg of dry weight and came from 78 mL (dry weight 0.38 g). Samples in the bottom part of the table (B section) contained 8.3 mg of dry weight and came from 100 mL (dry weight 0.27 g) of batch cultures growing on CDM with 333 mM glucose. The intracellular volume was calculated using the intracellular volume of E. coli (2.15 mL g^-1 dry weight) [37].

ISSN: 1475-2859