Development of renewable energy is rapidly being embraced by our society and industry to achieve the nation's economic growth goals and to help address the world's energy and global warming crises. Currently most of the bioethanol production is from the fermentation of agricultural crops and residues. There is much debate concerning the cost effectiveness and energy efficiency of such biomass based ethanol production processes. Here, we report the creation of a Synechocystis sp. PCC 6803 strain that can photoautotrophically convert CO 2 to bioethanol. Transformation was performed using a double homologous recombination system to integrate the pyruvate decarboxylase (pdc) and alcohol dehydrogenase II (adh) genes from obligately ethanol producing Zymomonas mobilis into the Synechocystis PCC 6803 chromosome under the control of the strong, light driven psbAII promoter. PCR based assay and ethanol production assay were used to screen for stable transformants. A computerized photobioreactor system was established for the experimental design and data acquisition for the analysis of the cyanobacterial cell cultures and ethanol production. The system described here shows an average yield of 5.2 mmol OD 730 unit À1 litre À1 day À1 with no required antibiotic/selective agent.
BackgroundThe yeast Saccharomyces cerevisiae is an important microorganism for both industrial processes and scientific research. Consequently, there have been extensive efforts to characterize its cellular processes. In order to fully understand the relationship between yeast's genome and its physiology, the stockpiles of diverse biological data sets that describe its cellular components and phenotypic behavior must be integrated at the genome-scale. Genome-scale metabolic networks have been reconstructed for several microorganisms, including S. cerevisiae, and the properties of these networks have been successfully analyzed using a variety of constraint-based methods. Phenotypic phase plane analysis is a constraint-based method which provides a global view of how optimal growth rates are affected by changes in two environmental variables such as a carbon and an oxygen uptake rate. Some applications of phenotypic phase plane analysis include the study of optimal growth rates and of network capacity and function.ResultsIn this study, the Saccharomyces cerevisiae genome-scale metabolic network was used to formulate a phenotypic phase plane that displays the maximum allowable growth rate and distinct patterns of metabolic pathway utilization for all combinations of glucose and oxygen uptake rates. In silico predictions of growth rate and secretion rates and in vivo data for three separate growth conditions (aerobic glucose-limited, oxidative-fermentative, and microaerobic) were concordant.ConclusionsTaken together, this study examines the function and capacity of yeast's metabolic machinery and shows that the phenotypic phase plane can be used to accurately predict metabolic phenotypes and to interpret experimental data in the context of a genome-scale model.
The pH‐responsive core‐shell microspheres of poly(styrene‐co‐methylacrylic acid) (PS‐co‐PMAA) containing a polystyrene (PS) core and a poly(methylacrylic acid) (PMAA) shell are synthesized by one‐stage soap‐free copolymerization and the catalyst system palladium‐iminodiacetic acid (IDA‐Pd) is immobilized on the outer shell‐layer of the core‐shell microspheres to form the quasi‐homogeneous and easily accessible catalyst PS‐co‐PMAA‐IDA‐Pd. This quasi‐homogeneous PS‐co‐PMAA‐IDA‐Pd catalyst is highly dispersed in the reaction medium just like a homogeneous one and can be separated like a heterogeneous catalyst by adjusting the pH of the reaction medium. Suzuki reactions employing the quasi‐homogeneous PS‐co‐PMAA‐IDA‐Pd catalyst are efficiently performed in water as the sole solvent under mild conditions such as room temperature. The PS‐co‐PMAA‐IDA‐Pd catalyst is also used in Heck reactions of a wide range of aryl halides with styrene and proves to be efficient in aqueous solution. The PS‐co‐PMAA‐IDA‐Pd catalyst has a low leaching loss and can be reused at least 4 times without loss of activity.
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