Sodium (Na+) concentration effects on metabolic pathway and estimation of ATP use in dark fermentation hydrogen production through stoichiometric analysis.

Batch experiments were conducted to investigate the effects of Na(+) concentration on hydrogen production with dark fermentation. The Na(+) concentration was varied from 0 to 8 g/L in the mixed culture using an anaerobic sludge treated by acid. The maximum hydrogen production was achieved with 1 g-Na(+)/L, whereas the hydrogen production was decreased over 2 g-Na(+)/L due to the inhibitory of Na(+). The mechanisms of Na(+) inhibition to the hydrogen production are studied using pure culture of Clostridium butyricum by calculating the energy balance. At a high sodium concentration, C. butyricum used a greater proportion of the ATP generated via fermentation for cell maintenance rather than for cell synthesis. Additionally, higher Na(+) concentrations shifted the fermentation process toward the acetate synthesis pathway instead of the butyrate pathway, and the value of Y(X/ATP) decreased. With high Na(+) concentrations, a greater ratio of hydrogen was produced via the oxidation of NADH. Excess hydrogen production decreased as the Na(+) concentration increased.

TRIAD1 is negatively regulated by the MDM2 E3 ligase.

Two RING fingers and DRIL1 (TRIAD1) is a proapoptotic protein that promotes p53 activation in several cancer cell lines, including MCF7, U2OS and A549 cells. In this study, we demonstrated that TRIAD1 is a novel ubiquitination target for proteasome-dependent degradation by murine double minute 2 (MDM2). TRIAD1 was found to interact with and be ubiquitinated by MDM2. RNA interference against MDM2 increased endogenous TRIAD1 protein stability. The functional study results suggested that TRIAD1 degradation by MDM2 suppresses TRIAD1-mediated cell growth. These data suggested a novel negative regulatory mechanism of TRIAD1 via MDM2 E3 ligase ubiquitination.

Effects of acid pre-treatment on bio-hydrogen production and microbial communities during dark fermentation.

Optimal conditions for acid pre-treatment were investigated for the enrichment of hydrogen-producing bacteria (HPB) in a mixed culture using three strong acids: HCl, HNO(3), and H2SO4 x HCl was selected as a suitable acid for the enrichment of HPB in the fermentation process. The volume of bio-hydrogen produced when the mixed culture was pre-treated using HCl at pH 2 was 3.2 times higher than that obtained without acid pre-treatment. Changes in the microbial community during acid pre-treatment were monitored using images obtained by the fluorescent in situ hybridization (FISH) method and the Live/Dead cell viability test. The tests clearly indicated that the Clostridium species of cluster I were the predominant strains involved in bio-H(2) fermentation, and could be selectively enriched by HCl pre-treatment.