The global pool of intracellular metabolites is a reflection of all the metabolic functions of an organism. In the absence of in situ methods capable of directly measuring metabolite pools, intracellular metabolite measurements need to be performed after an extraction procedure. In this study, we evaluated the optimization of technologies for generation of a global metabolomics profile for intracellular metabolites in Klebsiella oxytoca. Intracellular metabolites of K. oxytoca were extracted at the early stationary phase using six different common extraction procedures, including cold methanol, boiling ethanol, methanol/chloroform combinations, hot water, potassium hydroxide, and perchloric acid. The metabolites were subsequently collected for further analysis, and intracellular metabolite concentration profiles were generated using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. During analysis, the stability of metabolites extracted using cold methanol was clearly higher than that obtained by other extraction methods. For the majority of metabolites, extracts generated in this manner exhibited the greatest recovery, with high reproducibility. Therefore, the use of cold ethanol was the best extraction method for attaining a metabolic profile. However, in another parallel extraction method, perchloric acid may also be required to maximize the range of metabolites recovered, particularly to extract glucose 1-phosphate and NADPH.

The efficiency of the bioconversion process and the achievable end-product concentration decides the economic feasibility of microbial 2,3-butanediol (2,3-BDO) production. In 2,3-BDO production, optimization of culture condition is required for cell growth and metabolism. Also, the pH is an important factor that influences microbial performance. For different microorganisms and substrates, it has been shown that the distribution of the metabolites in 2,3-BDO fermentation is greatly affected by pH, and the optimum pH for 2,3-BDO production seems dependently linked to the particular strain and the substrate employed. Quantification analysis of intracellular metabolites and metabolic flux analysis (MFA) were used to investigate the effect of pH on the Klebsiella oxytoca producing 2,3-BDO and other organic acids. The main objectives of MFA are the estimation of intracellular metabolic fluxes and the identification of rate-limiting step and the key enzymes. This study was conducted under continuous aerobic conditions at different dilution rates (0.1, 0.2, and 0.3 h(-1)) and different pH values (pH 5.5 and 7.0) for the steady-state experimental data. In order to obtain the flux distribution, the extracellular specific rates were calculated from the experimental data using the metabolic network model of K. oxytoca. Intracellular metabolite concentration profiles were generated using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.

This study investigated the quality characteristics between spring cultivars of Kimchi cabbage(Brassica rapa L. ssp. pekinesis). We measured the weight, length, width, formation index, midrib thickness and moisture contents of fresh Kimchi cabbage for characteristics of growth. And we analyzed the free sugar, amino acid, organic acid, mineral, pectin and cellulose contents of fresh Kimchi cabbage. The hardness and firmness were measured for texture of fresh Kimchi cabbage. The weight of 'K-power' cultivar was the highest than other cultivars. The 'Jeongsang' cultivars was the thinnest midrib thickness in cultivars, but it was not significantly different. The free sugar levels of spring cultivars was the highest in 'Chunkwang'. Malic acid content of 'Jeongsang' was significantly different among spring cultivars. Also amino acid content of 'K-power' and 'Chunkwang' were significantly different among spring cultivars Mineral content in 'K-power' and 'Bomaknorang' were greater than that of other cultivars. Especially Na, Ca and Mg contents of 'K-power' and 'Bomaknorang' were higher four times than that of other cultivars. Pectin content of 'Jeongsang' was higher than that of other cultivars, but it was not significantly different. Hardness and Firmness were not significantly different among spring cultivars.