Qingdao Energy Research Institute, etc. developed high CO2 tolerant industrial oil-producing microalgae

Industrial microalgae can directly convert sunlight and flue gas into biodiesel, so it is one of the important measures to deal with global warming. However, the high concentration of CO2 in flue gas and the resulting acidic culture conditions often inhibit the growth of microalgae. Therefore, improving CO2 tolerance is one of the key bottlenecks in the design and construction of super photosynthetic carbon-fixing cell factories. Recently, the Single Cell Center of the Qingdao Institute of Bioenergy and Process Research, Chinese Academy of Sciences, by reversing the evolutionary clockwise research idea, was the first to elucidate the mechanism of industrial microalgae responding to high concentrations of CO2, and developed a high CO2 tolerant industrial oil-producing microalgal cell factory . The result was published online at Metabolic Engineering on March 21.

Greenhouse gases such as CO2 emitted by human activities have caused major environmental and social problems such as global climate change and ocean acidification. Using industrial oil-producing microalgae to directly convert industrial CO2 emission sources such as flue gas into advanced biofuels such as diesel fuel is of great significance for reducing greenhouse gas emissions and curbing global warming. Marine phytoplankton, including microalgae, adapt to the current CO2 content of 0.04% of the Earth ’s atmosphere, and fix 40% of the global fixed amount of CO2 every year. However, the CO2 content in the flue gas is higher than 5%, which is more than 100 times the atmospheric carbon content. The resulting acidification of the culture environment, while reducing the occurrence of biological pollution, also generally inhibits the growth and reproduction of industrial oil-producing microalgae, thereby greatly reducing the economics of the industrial biological carbon-fixing oil production process. Nannochloropsis spp. Is an industrial oil-producing microalgae that can be cultivated outdoors on a large scale around the world. They have the outstanding advantages of fast growth speed, strong carbon dioxide tolerance, both seawater and freshwater cultivation, and perfect genetic operation. Researchers such as Wei Li from the Single Cell Center of Qingdao Energy Institute have proposed scientific assumptions that their use and tolerance of CO2 are related to the Carbon Concentrating Mechanism (CCM). First, using system biology ideas, combined with research methods such as subcellular localization, a key target related to high CO2 stress was discovered, namely a special carbonic anhydrase (CA2) located in the cytoplasm. Compared with 5% CO2 culture, CA2 is specifically activated at very low CO2 concentration, so it is a key gene for the CCM system to sense and respond to CO2 concentration in the environment.

Furthermore, the researchers proposed that since CCM is an evolutionary result of algae gradually adapting to the current atmosphere (low-concentration CO2 environment) from the ancient atmosphere (high-concentration CO2 environment), if artificially destroying or inhibiting CCM activity, can it "reverse the clock of evolution" , Artificially realize the "return to ancestor" of industrial microalgae, so as to restore its adaptability to high concentration of CO2? The experiment proves that under 5% CO2, the engineered Nannochloropsis strains targeting CA2 knockdown can increase the biomass yield by more than 30%, and the oil content is not affected. This excellent trait can be exhibited in various types of light cultivation facilities and cultivation scales at various spatial scales, and has considerable genetic stability. Further research found that knocking down CA2 significantly improved the intracellular pH microenvironment, thereby alleviating the toxic effects of extracellular hydrogen ions on cells and ultimately maintaining biomass growth (see figure). Interestingly, the growth advantage of the engineered algae strain is only exhibited under the flue gas culture conditions. If the air concentration is CO2, the engineered algae strain loses its growth advantage. Therefore, this study not only proved that the adaptability of industrial microalgae CO2 content can be rationally regulated, but also invented an original ecological control strategy for engineering algae strains.

How to improve the CO2 utilization efficiency of food and energy crops has always been the goal of the industry. Existing work usually takes the promotion and improvement of CCM activity as the core idea to improve the carbon sequestration efficiency of crops. The study "reverses the way" and proposes that by reversing the evolution of CCM and inhibiting its activity, it can increase the yield of crops under high CO2 conditions.

This work was presided over by Xu Jian, a researcher at the Single Cell Research Center of the Qingdao Energy Institute and Ansgar Poetsch, a professor at the Ruhr University in Germany, with the help of Hu Qiang and Hu Hanhua, researchers at the Institute of Hydrobiology, Chinese Academy of Sciences. The research was supported by the CO2 key deployment project of the Chinese Academy of Sciences, the “One Three Five” project of the Institute and the National Natural Science Foundation of China.

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