Advanced Imaging Technologies to Empower Bioenergy Discoveries

Abstract: Plants and microbes are major organisms engineered for the manufacturing renewable fuels and chemicals to replace petroleum. However, in vivo quantification of target molecule production is highly challenging, which hinders the development of high efficient systems. In particular, the 3-D, in vivo, and real-time quantification of chemical species represents a major challenge. Recent advancements of CARS and SRS technologies offered unique solutions for 3-D in vivo, realtime imaging. We first utilized SRS to carry out thorough analysis of an engineered subcellular organelle for hydrocarbon. Photosynthetic terpene production represents one of the most energy and carbon efficient biological routes for reducing CO2 to hydrocarbons. We carried out thorough pathway and organelle engineering to improve squalene accumulation in tobacco. In particular, we have modified the animal oleosin protein and engineered it into the tobacco leaves to produce subcellular oil droplet to enhance squalene production.  The SRS imaging revealed clear formation of droplets in plant leaves. Furthermore, the chemical species identification indicated that the droplets contains squalene. The imaging results well correlates with the GC/MS results, highlighting four times increase of squalene production in the engineered plants with squalene droplets. Furthermore, we have engineered cyanobacteria to produce limonene at high productivity. Using SRS, we were able to identify the hydrophobic interact among the cells caused by limonene, which further leads to the development of auto-sediment-based low-cost harvesting technology. Using this technology, we were able to develop a semi-continuous cultivation to achieve highest reported outdoor algal productivity. Furthermore, we have used Raman imaging to quantitively evaluate the beta-carotenoid changes during plant stress responses. Furthermore, in our latest efforts, we are exploring the possibility of using QUIP and QGI to image plant samples for studying photosynthesis process, especially during dark to light transition. Overall, the cutting-edge imaging technology has substantially improved the fundamental understanding and technology development in bioproduction.

Bio: Joshua Yuan serves as the Lucy & Stanley Lopata Professor, the Chair for the Department of Energy, Environmental and Chemical Engineering, and the Director of Carbon Utilization Redesign through Biomanufacturing (CURB) Engineering Research Center at Washington University in St. Louis. Previously, he was a faculty member at Texas A&M University since 2008 and was appointed as the Chair for Synthetic Biology and Renewable Products in 2018. He has served as the Director of the Synthetic and Systems Biology Innovation Hub since 2015. Joshua has built a career in developing, disseminating, and implementing sustainability solutions for the future. His research focuses on carbon capture and utilization, renewable biomaterials, biomass processing, and biorefining, as well as synthetic and systems biology. His group has developed various routes for carbon capture and conversion, including the algae-based and electro-microbial hybrid processes. In particular, his team achieved the highest reported algal outdoor productivity through artificial intelligence-guided process design and microbial engineering. He has been awarded four U.S. patents and has five pending. He has written more than 100 peer-reviewed journal articles, published in Joule, Chem, Matter, Nature Communications, Green Chemistry, Advanced Sciences, and PNAS, among others. He has won numerous awards and honors, including the Excellence in Innovation Award from Texas A&M University System in 2017 and the Gamma Sigma Delta Outstanding Graduate Student Award in 2007, among others. He is now a Fellow of the Royal Society of Chemistry.

Advanced Imaging Technologies to Empower Bioenergy Discoveries

Abstract: Plants and microbes are major organisms engineered for the manufacturing renewable fuels and chemicals to replace petroleum. However, in vivo quantification of target molecule production is highly challenging, which hinders the development of high efficient systems. In particular, the 3-D, in vivo, and real-time quantification of chemical species represents a major challenge. Recent advancements of CARS and SRS technologies offered unique solutions for 3-D in vivo, realtime imaging. We first utilized SRS to carry out thorough analysis of an engineered subcellular organelle for hydrocarbon. Photosynthetic terpene production represents one of the most energy and carbon efficient biological routes for reducing CO2 to hydrocarbons. We carried out thorough pathway and organelle engineering to improve squalene accumulation in tobacco. In particular, we have modified the animal oleosin protein and engineered it into the tobacco leaves to produce subcellular oil droplet to enhance squalene production.  The SRS imaging revealed clear formation of droplets in plant leaves. Furthermore, the chemical species identification indicated that the droplets contains squalene. The imaging results well correlates with the GC/MS results, highlighting four times increase of squalene production in the engineered plants with squalene droplets. Furthermore, we have engineered cyanobacteria to produce limonene at high productivity. Using SRS, we were able to identify the hydrophobic interact among the cells caused by limonene, which further leads to the development of auto-sediment-based low-cost harvesting technology. Using this technology, we were able to develop a semi-continuous cultivation to achieve highest reported outdoor algal productivity. Furthermore, we have used Raman imaging to quantitively evaluate the beta-carotenoid changes during plant stress responses. Furthermore, in our latest efforts, we are exploring the possibility of using QUIP and QGI to image plant samples for studying photosynthesis process, especially during dark to light transition. Overall, the cutting-edge imaging technology has substantially improved the fundamental understanding and technology development in bioproduction.

Bio: Joshua Yuan serves as the Lucy & Stanley Lopata Professor, the Chair for the Department of Energy, Environmental and Chemical Engineering, and the Director of Carbon Utilization Redesign through Biomanufacturing (CURB) Engineering Research Center at Washington University in St. Louis. Previously, he was a faculty member at Texas A&M University since 2008 and was appointed as the Chair for Synthetic Biology and Renewable Products in 2018. He has served as the Director of the Synthetic and Systems Biology Innovation Hub since 2015. Joshua has built a career in developing, disseminating, and implementing sustainability solutions for the future. His research focuses on carbon capture and utilization, renewable biomaterials, biomass processing, and biorefining, as well as synthetic and systems biology. His group has developed various routes for carbon capture and conversion, including the algae-based and electro-microbial hybrid processes. In particular, his team achieved the highest reported algal outdoor productivity through artificial intelligence-guided process design and microbial engineering. He has been awarded four U.S. patents and has five pending. He has written more than 100 peer-reviewed journal articles, published in Joule, Chem, Matter, Nature Communications, Green Chemistry, Advanced Sciences, and PNAS, among others. He has won numerous awards and honors, including the Excellence in Innovation Award from Texas A&M University System in 2017 and the Gamma Sigma Delta Outstanding Graduate Student Award in 2007, among others. He is now a Fellow of the Royal Society of Chemistry.