Studies on the production of branched-chain alcohols in engineered Ralstonia eutropha

Wild-type Ralstonia eutropha H16 produces polyhydroxybutyrate (PHB) as an intracellular carbon storage material during nutrient stress in the presence of excess carbon. In this study, the excess carbon was redirected in engineered strains from PHB storage to the production of isobutanol and 3-methyl-1-butanol (branched-chain higher alcohols). These branched-chain higher alcohols can directly substitute for fossil-based fuels and be employed within the current infrastructure. Various mutant strains of R. eutropha with isobutyraldehyde dehydrogenase activity, in combination with the overexpression of plasmid-borne, native branched-chain amino acid biosynthesis pathway genes and the overexpression of heterologous ketoisovalerate decarboxylase gene, were employed for the biosynthesis of isobutanol and 3-methyl-1-butanol. Production of these branched-chain alcohols was initiated during nitrogen or phosphorus limitation in the engineered R. eutropha. One mutant strain not only produced over 180 mg/L branched-chain alcohols in flask culture, but also was significantly more tolerant of isobutanol toxicity than wild-type R. eutropha. After the elimination of genes encoding three potential carbon sinks (ilvE, bkdAB, and aceE), the production titer improved to 270 mg/L isobutanol and 40 mg/L 3-methyl-1-butanol. Semicontinuous flask cultivation was utilized to minimize the toxicity caused by isobutanol while supplying cells with sufficient nutrients. Under this semicontinuous flask cultivation, the R. eutropha mutant grew and produced more than 14 g/L branched-chain alcohols over the duration of 50 days. These results demonstrate that R. eutropha carbon flux can be redirected from PHB to branched-chain alcohols and that engineered R. eutropha can be cultivated over prolonged periods of time for product biosynthesis.United States. Dept. of EnergyUnited States. Advanced Research Projects Agency-Energ

How Well Can Aerosol Measurements from the Terra Morning Polar Orbiting Satellite Represent the Daily Aerosol Abundance and Properties?

The Terra mission, launched at the dawn of 1999, and Aqua mission to be launched soon, will possess innovative measurements of the aerosol daily spatial distribution, distinguish between dust, smoke and regional pollution and measure aerosol radiative forcing of climate. Their polar orbit gives daily global coverage, however measurements are acquired at specific time of the day. To what degree can present measurements from Terra taken between 10:00 and 11:30 AM local time, represent the daily average aerosol forcing of climate? Here we answer this question using 7 years of data from the distributed ground based 50-70 instrument Aerosol Robotic Network (AERONET) This (AERONET) half a million measurement data set shows that Terra aerosol measurements represent the daily average values within 5%. The excellent representation is found for large dust particles or small aerosol particles from Fires or regional pollution and for any range of the optical thickness, a measure of the amount of aerosol in the atmosphere