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Acetic acid is an abundant material that can be used as a carbon source by microorganisms. Despite its abundance, its toxicity and low energy content make it hard to utilize as a sole carbon source for biochemical production. To increase acetate utilization and isobutanol production with engineered Escherichia coli, the feasibility of utilizing acetate and metabolic engineering was investigated. The expression of acs, pckA, and maeB increased isobutanol production by up to 26%, and the addition of TCA cycle intermediates indicated that the intermediates can enhance isobutanol production. For isobutanol production from acetate, acetate uptake rates and the NADPH pool were not limiting factors compared to glucose as a carbon source. This work represents the first approach to produce isobutanol from acetate with pyruvate flux optimization to extend the applicability of acetate. This technique suggests a strategy for biochemical production utilizing acetate as the sole carbon source.

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A field experimental study was performed during the growing season of 2001 to evaluate water and nutrient balances in paddy rice culture. Three plots of standard fertilization (SF), excessive fertilization (EF, 150% of SF), and reduced fertilization (RF, 70% of SF) were used and the size of treatment plot was 3,000 m 2 , respectively. The hydrologic and water quality was field monitored throughout the crop stages. The water balance analyses indicated that approximately half (47-54%) of the total outflow was lost through surface drainage, with the remainder consumed by evapotranspiration. Statistical analysis showed that there was no significant effect of fertilization rates on nutrient outflow through the surface drainage or rice yield. Reducing fertilization of rice paddy may not work well to mitigate the non-point source nutrient loading in the range of normal farming practices. Instead, the reduction in surface drainage could be important to controlling the loading. Suggestive measures that may be applicable to reduce surface drainage and nutrient losses include water-saving irrigation by reducing ponded water depth, raising the weir height in diked rice fields, and minimizing forced surface drainage as recommended by other researchers. The suggested practices can cause some deviations from conventional farming practices, and further investigations are recommended.

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Shewanella oneidensis MR-1 is one of the most well-known metal-reducing bacteria and it has been extensively studied for microbial fuel cell and bioremediation aspects. In this study, we have examined S. oneidensis MR-1 as an isobutanol-producing host by assessing three key factors such as isobutanol synthetic genes, carbon sources, and electron supply systems. Heterologous Ehrlich pathway genes, kivD encoding ketoisovalerate decarboxylase and adh encoding alcohol dehydrogenase, were constructed in S. oneidensis MR-1. Among the composition of carbon sources examined, 2% of N-acetylglucosamine, 1.5% of pyruvate and 2% of lactate were found to be the most optimal nutrients and resulted in 10.3 mg/L of isobutanol production with 48 h of microaerobic incubation. Finally, the effects of metal ions (electron acceptor) and direct electron transfer systems on isobutanol production were investigated, and Fe(2+) ions increased the isobutanol production up to 35%. Interestingly, deletion of mtrA and mtrB, genes responsible for membrane transport systems, did not have significant impact on isobutanol production. Finally, we applied engineered S. oneidensis to a bioelectrical reactor system to investigate the effect of a direct electron supply system on isobutanol production, and it resulted in an increased growth and isobutanol production (up to 19.3 mg/L). This report showed the feasibility of S. oneidensis MR-1 as a genetic host to produce valuable biochemicals and combine an electron-supplying system with biotechnological applications.

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