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A brief background

    
    Global climate change poses a significant threat to human society, and more and more countries are elevating "carbon neutrality" as a national strategy, proposing a vision for a carbon free future. As the world's largest developing country and largest coal consumer, China has proposed the "dual carbon" goal to achieve carbon peak as soon as possible and work together with other countries to achieve net zero CO2 emissions by around the middle of this century.
   In 2017, greenhouse gas emissions from commercial flights in the global civil aviation industry accounted for over 96% of the industry's total emissions, becoming the largest source of emissions for the civil aviation industry. The basic emissions of CO2 generated by the combustion of aviation coal are releasing in the atmospheric stratosphere, and its greenhouse effect harm is far greater than that of other industries. At the same time, aviation transportation has strong international attributes, making it one of the focuses of global climate change response and facing severe challenges in reducing greenhouse gas emissions.
    So, we seek to produce an environmentally friendly and sustainable biofuel through synthetic biology technology. Initially, we planned to use straw as raw material for zero carbon production of biofuels and maximize the substitution of chemical fuels. Ultimately, we hope to achieve the goal of optimizing the World energy resources situation!
    As a new discipline, synthetic biology can always solve the practical problems around us with the most advanced technology, the lowest cost and the highest efficiency, and bring infinite possibilities for future human life.

Our team put stress on using biology to synthesize Isobutanol

    
    Biofuels (ethanol, butanol, Isobutanol, etc.) are a kind of renewable energy with great potential. Compared with fossil fuels, biofuels are mainly produced from renewable biomass resources, so CO2 generated by combustion will not increase emissions, and can even reduce the net emissions of greenhouse gases. All major economies in the world regard the development of synthetic Bioenergy as an important strategic choice to ensure energy security, environmental quality and economic development. Among all kinds of biofuels, Isobutanol, as a branched chain tetracarbyl alcohol, has the advantages of high heat density, high combustion value, low volatility and low hygroscopicity compared with traditional Ethanol fuel and n-butanol, and is regarded as a higher performance biofuel that can replace ethanol. Isobutanol can also be used as aviation fuel to promote aerospace development.

Taking Isobutanol as the main line, our therapy consists of three parts

    
    Zymomonas mobilis is a GRAS safe strain that naturally produces ethanol. It is currently the only known microorganism that can utilize the Entner-Doudoroff(ED) pathway to metabolize ethanol under anaerobic conditions. It has advantages such as fast glucose metabolism, high ethanol yield (98%), strong ethanol tolerance (15% v/v), small genome size, and complete genetic manipulation tools.
   In the first section:By introducing the five acting genes Bsals, ilvC, ilvD, kdcA, adhA in the Isobutanol biosynthesis pathway into the Z. mobilis, and adopting the corresponding static transformation strategies, including optimizing the plasmid; Increase the copy of kdcA to improve the stability of gene expression; Using dcpf1 to inhibit the expression of pdc genes in the ethanol production pathway using CRISPRi; Cofactor balance strategy, we have strengthened the biosynthesis path of Isobutanol to achieve high yield of aviation fuel Isobutanol.
   In the second section:We introduced PHB pathway related genes PhbCAB, zwf and EUP pathway genes into the optimized strain for Isobutanol production, so that PHB can be co produced while Isobutanol production is high, increasing part of the income.
   In the last section:In order to fix CO2 during metabolism, we introduced the carbon dioxide fixation pathway Calvin cycle, and introduced carboxylated somatic cell apparatus to assist carbon dioxide fixation. Ultimately, we achieved zero carbon production of Isobutanol and PHB.

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