Kinetic modeling and sensitivity analysis of acetone-butanol-ethanol production.

A kinetic simulation model of metabolic pathways that describes the dynamic behaviors of metabolites in acetone-butanol-ethanol (ABE) production by Clostridium saccharoperbutylacetonicum N1-4 was proposed using a novel simulator WinBEST-KIT. This model was validated by comparing with experimental time-course data of metabolites in batch cultures over a wide range of initial glucose concentrations (36.1-295 mM). By introducing substrate inhibition, product inhibition of butanol, activation of butyrate and considering the cessation of metabolic reactions in the case of insufficiency of energy after glucose exhaustion, the revised model showed 0.901 of squared correlation coefficient (r(2)) between experimental time-course of metabolites and calculated ones. Thus, the final revised model is assumed to be one of the best candidates for kinetic simulation describing dynamic behavior of metabolites in ABE production. Sensitivity analysis revealed that 5% increase in reaction of reverse pathway of butyrate production (R(17)) and 5% decrease in reaction of CoA transferase for butyrate (R(15)) highly contribute to high production of butanol. These system analyses should be effective in the elucidation which pathway is metabolic bottleneck for high production of butanol.

Novel high-efficient butanol production from butyrate by non-growing Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564) with methyl viologen.

Non-growing Clostridium saccharoperbutylacetonicum N1-4 hardly produced butanol from only butyrate. As adding glucose to the medium, butyrate utilization and butanol production were stimulated. Addition of 0.1 mM methyl viologen as electron carrier resulted in the highest yield of butanol of 0.671 mol/mol to butyrate and glucose.

Accumulation and pharmacokinetics of estrogenic chemicals in the pre- and post-hatch embryos of the frog Rana rugosa.

Amphibian eggs spawned in water are exposed immediately to various chemicals present in their water. The present study aimed to investigate the accumulation and pharmacokinetics of 17α-ethynylestradiol (EE(2)), bisphenol A (BPA), and nonylphenol (NP), as well as 17ß-estradiol (E(2)), in the pre-hatch and post-hatch embryos of the frog Rana rugosa. Fertilized eggs were exposed to chemicals at a final concentration of 500 nM in breeding water for two days, then the embryos with jelly coats were reared in fresh-breeding water without supplementation of the xenoestrogens for six more days. All exogenous chemicals were concentrated in the embryo body at two days after fertilization, whereas their concentrations in the jelly coat were the same as those in the breeding water. The bioconcentration factors for E(2), EE(2), BPA, and NP were 217.9, 170.2, 382.3, and 289.1, respectively, suggesting that the estrogenic chemicals were concentrated in the embryo body through the jelly coat.

Development of high-speed and highly efficient butanol production systems from butyric acid with high density of living cells of Clostridium saccharoperbutylacetonicum.

Living cells are alive and have the butanol-producing ability but not much proliferation under nitrogen source-limited condition. We investigated various butanol production systems with high density of living cells of Clostridium saccharoperbutylacetonicum N1-4 supplemented with methyl viologen (MV) as an electron carrier and nutrient dosing for activity regeneration. In continuous butanol production with high density of living cells, butanol yield was drastically increased from 0.365 C-mol/C-mol with growing cells to 0.528 C-mol/C-mol at a dilution rate of 0.85 h⁻¹, being increased with the butanol to total solvent ratio. This yield was increased to 0.591 C-mol/C-mol by adding 0.01 mM MV. MV addition increased not only butanol yield but also butanol concentration and productivity as compared to those without MV addition. However, living cells lost their activity with incubation time, which lowered the operational stability of the system. Therefore, to maintain constant stability, activity regeneration was carried out with high density of living cells and MV. This system produced butanol at high concentration (9.40 g l⁻¹) and productivity (7.99 g l⁻¹ h⁻¹) for approximately 100 h with maintenance of considerably high yield of butanol (0.686 C-mol/C-mol). Thus, we established a high-speed and highly efficient butanol production system.

Kinetic modeling and sensitivity analysis of xylose metabolism in Lactococcus lactis IO-1.

We proposed a kinetic simulation model of xylose metabolism in Lactococcus lactis IO-1 that describes the dynamic behavior of metabolites using the simulator WinBEST-KIT. This model was developed by comparing the experimental time-course data of metabolites in batch cultures grown in media with initial xylose concentrations of 20.3-57.8 g/l with corresponding calculated data. By introducing the terms of substrate activation, substrate inhibition, and product inhibition, the revised model showed a squared correlation coefficient (r2) of 0.929 between the experimental time-course of metabolites and the calculated data. Thus, the revised model is assumed to be one of the best candidates for kinetic simulation describing the dynamic behavior of metabolites. Sensitivity analysis revealed that pyruvate flux distribution is important for higher lactate production. To confirm the validity of our kinetic model, the results of the sensitivity analysis were compared with enzyme activities observed during increasing lactate production by adding natural rubber serum powder to the xylose medium. The experimental results on pyruvate flux distribution were consistent with the prediction by sensitivity analysis.