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BackgroundProduction of fuels from the abundant and wasteful CO2 is a promising approach to reduce carbon emission and consumption of fossil fuels. Autotrophic microbes naturally assimilate CO2 using energy from light, hydrogen, and/or sulfur. However, their slow growth rates call for investigation of the possibility of heterotrophic CO2 fixation. Although preliminary research has suggested that CO2 fixation in heterotrophic microbes is feasible after incorporation of a CO2-fixing bypass into the central carbon metabolic pathway, it remains unclear how much and how efficient that CO2 can be fixed by a heterotrophic microbe.ResultsA simple metabolic flux index was developed to indicate the relative strength of the CO2-fixation flux. When two sequential enzymes of the cyanobacterial Calvin cycle were incorporated into an E. coli strain, the flux of the CO2-fixing bypass pathway accounts for 13 % of that of the central carbon metabolic pathway. The value was increased to 17 % when the carbonic anhydrase involved in the cyanobacterial carbon concentrating mechanism was introduced, indicating that low intracellular CO2 concentration is one limiting factor for CO2 fixation in E. coli. The engineered CO2-fixing E. coli with carbonic anhydrase was able to fix CO2 at a rate of 19.6 mg CO2 L−1 h−1 or the specific rate of 22.5 mg CO2 g DCW−1 h−1. This CO2-fixation rate is comparable with the reported rates of 14 autotrophic cyanobacteria and algae (10.5–147.0 mg CO2 L−1 h−1 or the specific rates of 3.5–23.7 mg CO2 g DCW−1 h−1).ConclusionsThe ability of CO2 fixation was created and improved in E. coli by incorporating partial cyanobacterial Calvin cycle and carbon concentrating mechanism, respectively. Quantitative analysis revealed that the CO2-fixation rate of this strain is comparable with that of the autotrophic cyanobacteria and algae, demonstrating great potential of heterotrophic CO2 fixation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-015-0268-1) contains supplementary material, which is available to authorized users.

Recycling of carbon dioxide (CO2) into fuels and chemicals is a potential approach to reduce CO2 emission and fossil-fuel consumption. Autotrophic microbes can utilize energy from light, hydrogen, or sulfur to assimilate atmospheric CO2 into organic compounds at ambient temperature and pressure. This provides a feasible way for biological production of fuels and chemicals from CO2 under normal conditions. Recently great progress has been made in this research area, and dozens of CO2-derived fuels and chemicals have been reported to be synthesized by autotrophic microbes. This is accompanied by investigations into natural CO2-fixation pathways and the rapid development of new technologies in synthetic biology. This review first summarizes the six natural CO2-fixation pathways reported to date, followed by an overview of recent progress in the design and engineering of CO2-fixation pathways as well as energy supply patterns using the concept and tools of synthetic biology. Finally, we will discuss future prospects in biological fixation of CO2.carbon dioxide fixation, synthetic biology, CO2-fixation pathway, energy supply Citation:

Natural CO 2 fixation cycles usually comprise multiple reactions, which may reduce the efficiency of the cycle. Here, we report the design and experimental demonstration of a minimized synthetic CO 2 fixation cycle which contains only four reactions. The cycle comprises pyruvate carboxylase, oxaloacetate acetylhydrolase, acetate-CoA ligase, and pyruvate synthase and is named the POAP cycle. The POAP cycle can condense two molecules of CO 2 into one molecule of oxalate in each step at the expense of two molecules of ATP and one reducing equivalent in the form of NAD(P)H. By identifying a ferredoxin from Hydrogenobacter thermophilus that can efficiently drive the rate-limiting reductive carboxylation step, the POAP cycle can be operated at 50 °C under anaerobic conditions, reaching a CO 2 fixation rate of 8.0 nmol CO 2 min −1 mg −1 CO 2 -fixing enzymes. The design and demonstration of the POAP cycle may provide a model to study CO 2 fixation in the earliest organisms.

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Epithelial-Mesenchymal Transition (EMT) is a critical step in the progression of cancer. Malignant melanoma, a cancer developed from pigmented melanocytes, metastasizes through an EMT-like process. Ten-eleven translocation (TET) enzymes, catalyzing the conversion of 5-methylcytosine (5mC) to 5-hydroxylmethylcytosine (5-hmC), are down regulated in melanoma. However, their roles in the progression and the EMT-like process of melanoma are not fully understood. Here we report that DNA methylation induced silencing of TET2 and TET3 are responsible for the EMT-like process and the metastasis of melanoma. TET2 and TET3 are down regulated in the TGF-β1-induced EMT-like process, and the knocking down of TET2 or TET3 induced this EMT-like process. A DNA demethylating agent antagonized the TGF-β-induced suppression of TET2 and TET3. Furthermore, a ChIP analysis indicated that enhanced recruitment of DNMT3A (DNA Methyltransferase 3A) is the mechanism by which TGF-β induces the silencing of TET2 and TET3. Finally, the overexpression of the TET2 C-terminal sequence partially rescues the TGF-β1-induced EMT-like process in vitro and inhibits tumor growth and metastasis in vivo. Hence, our data suggest an epigenetic circuitry that mediates the EMT activated by TGF-β. As an effector, DNMT3A senses the TGF-β signal and silences TET2 and TET3 promoters to induce the EMT-like process and metastasis in melanoma.

Targeting mitotic kinase monopolar spindle 1 (Mps1) for tumor therapy has been investigated for many years. Although it was suggested that Mps1 regulates cell viability through its role in spindle assembly checkpoint (SAC), the underlying mechanism remains less defined. In an endeavor to reveal the role of high levels of mitotic kinase Mps1 in the development of colon cancer, we unexpectedly found the amount of Mps1 required for cell survival far exceeds that of maintaining SAC in aneuploid cell lines. This suggests that other functions of Mps1 besides SAC are also employed to maintain cell viability. Mps1 regulates cell viability independent of its role in cytokinesis as the genetic depletion of Mps1 spanning from metaphase to cytokinesis affects neither cytokinesis nor cell viability. Furthermore, we developed a single-cycle inhibition strategy that allows disruption of Mps1 function only in mitosis. Using this strategy, we found the functions of Mps1 in mitosis are vital for cell viability as short-term treatment of mitotic colon cancer cell lines with Mps1 inhibitors is sufficient to cause cell death. Interestingly, Mps1 inhibitors synergize with microtubule depolymerizing drug in promoting polyploidization but not in tumor cell growth inhibition. Finally, we found that Mps1 can be recruited to mitochondria by binding to voltage-dependent anion channel 1 (VDAC1) via its C-terminal fragment. This interaction is essential for cell viability as Mps1 mutant defective for interaction fails to main cell viability, causing the release of cytochrome c. Meanwhile, deprivation of VDAC1 can make tumor cells refractory to loss of Mps1-induced cell death. Collectively, we conclude that inhibition of the novel mitochondrial function Mps1 is sufficient to kill tumor cells.

China initiated its acetone-butanol-ethanol (ABE) industry in the 1950s; it peaked in the 1980s, and ended at the end of the last century owing to the development of more competitive petrochemical pathways. However, driven by the high price of crude oil and environmental concerns raised by the over-consumption of petrochemical products, biofuels and bio-based chemicals including butanol have garnered global attention again. Currently, butanol produced from ABE fermentation is mainly used as an industrial solvent or a platform chemical for several bulk derivatives, and is also believed to be a potential biofuel. A number of plants have been built or rebuilt in recent years in China for butanol production with the ABE process. Chinese researchers also show great interest in the improvement of the production strains and corresponding processes. They have applied conventional mutagenesis methods to improve butanol-producing strains such as the Clostridium acetobutylicum mutant strains EA2018 (butanol ratio of 70%) and Rh8 (butanol tolerance of 19 g/L). The omics technologies, such as genome sequencing, proteomic and transcriptomic analysis, have been adapted to elucidate the characteristics of different butanol-producing bacteria. Based on the group II intron method, the genetic manipulation system of C. acetobutylicum was greatly improved, and some successful engineering strains were developed. In addition, research in China also covers the downstream processes. This article reviews up-to-date progress on biobutanol production in China.