Metabolome analysis of Escherichia coli ATCC25922 cells treated with high hydrostatic pressure at 400 and 600 MPa

Abstract

Escherichia coli cells were treated with high hydrostatic pressure (HHP) at 400 and 600MPa. Metabolites (70-1027m/z) extracted from HHP-treated cells were analyzed using capillary electrophoresis-time-of-flight mass spectrometry and were compared with those extracted from control cells (not treated with HHP). A total of 133 metabolites were identified and mapped to metabolic pathways, and many of these (42.1%) decreased due to the HHP treatment, including NAD+, NADP+, ATP, and substrates for DNA synthesis. Principal component analysis suggested that the central sugar and nucleic acid metabolic pathways were strongly influenced by HHP. A bottleneck in the central sugar metabolic pathway was observed in HHP-treated cells, which created a metabolic imbalance; metabolites mapped upstream (glucose 6-phosphate, fructose 6-phosphate, and fructose 1,6-diphosphate) were accumulated and those downstream (3-phosphoglycerate, 2-phosphoglycerate, and phosphoenolpyruvate) were depleted. Ribonucleotides were decreased, but the reduction was moderate compared with that of substrates for DNA synthesis; the exception was ATP, which also substantially decreased. The bottleneck in the glycolytic pathway partly explained the exhaustion of ATP. NAD+/NADH ratio of HHP treated cells was comparable with that of untreated control cells.