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To investigate the relationship between the production of poly(3-hydroxybutyrate) (PHB) and metabolic changes during different growth phases, a non-sterile batch fermentation process involving an alkalophilic and halophilic bacterium, Halomonas sp. KM-1, was used. Intracellular metabolites were analyzed using gas chromatography-mass spectrometry to characterize the metabolic profile. Significant changes relating to PHB production were observed in the TCA cycle, lipid-synthesis and amino acid biosynthetic pathways were found to shift dramatically between the exponential growth and stationary phases. During the stationary phase, 17 metabolites were upregulated and a cell dry mass of 17.8 g/L that included 44.8% PHB was observed at 24h in 5% glucose-supplemented cultures, whereas 11 metabolites were upregulated and a cell dry mass of 38.4 g/L that included 73.7% PHB was observed at 36 h in 10% glucose-supplemented cultures. This study provides pattern analysis of metabolite regulation during PHB accumulation, indicating that multicomponent and phase-specific mechanisms are involved.

A robust and convenient sheathless CE/ESI-MS interface realized with an ionophore membrane-packed electro-conduction channel is described. Sheathless interfaces that may provide higher sensitivity for MS detection than sheath flow-supported interfaces generally show instability and short lifetimes due to their imperfection in making an electrical contact with the emitter tip. In this work, we designed a sheathless interface based on an ionophore membrane-packed electro-conduction channel. At the joining point of the CE capillary and the emitter capillary, the conduction channel was implemented toward the exterior of the interface body, where a platinum wire electrode was placed. The conduction channel transferred the electric field from the external Pt electrode to the joining point, but prevented the effluent of CE from leaking. The interface body was designed to have receptacles for standard capillary tubing with finger-tight fittings, which allowed easy replacement of capillary tubing. Stable electrospray was observed for an extended time period without any signs of bubbling or damage to the emitter tip. No significant increment of dead-volume at the interface was observed for well-aligned capillaries. Sensitive and stable CE-MS detection of the model compound of creatinine and uric acid was demonstrated.