Park, Sujeong Park, Bo-ram Jeong, Deokyeol Park, Jongbeom Ko, Ja Kyong Kim, Soo-Jung Kim, Jeong-Sun Jin, Yong-Su Kim, Soo Rin 2024-01-12T06:32:31Z 2024-01-12T06:32:31Z 2023-11-21 2023-11 1359-5113 https://pubs.kist.re.kr/handle/201004/79753 Industrial biotechnology based on yeast fermentation is a promising strategy that can alleviate global warming and climate change. However, Saccharomyces cerevisiae, widely used in bioprocesses, releases a large amount of carbon dioxide (CO2) during fermentation. This study developed a mixotrophic CO2-fixing S. cerevisiae to achieve carbon neutrality and sustainability in bioprocess. A CO2-fixation pathway was constructed in a xylose-utilizing S. cerevisiae by heterologous expression of ribulose-1,5-bisphosphate carboxylase-oxygenase (RuBisCO) and phosphoribulokinase (PRK). Furthermore, a delta-integration strategy was utilized, and the RuBisCO gene copy number was increased to 10 copies to improve the efficiency of CO2-fixation. An additional Cas9-based genome editing was performed to overexpress other CO2-fixation related genes. The resulting CO2-fixing yeast, SJ03, exhibited the highest RuBisCO activity. During anaerobic xylose fermentation, ethanol concentration was increased by 17% and ethanol yield was increased by 16% compared to the control strain. In addition, CO2 emissions decreased by 7%. These results suggest that overexpression of the CO2-fixation pathway coupled with xylose utilization in S. cerevisiae might reduce CO2 emission in bioprocesses. English Elsevier Applied Science Functional expression of RuBisCO reduces CO2 emission during fermentation by engineered Saccharomyces cerevisiae Article 10.1016/j.procbio.2023.10.013 1 Process Biochemistry, v.134, no.Part 1, pp.286 - 293 Process Biochemistry 134 Part 1 286 293 N scie scopus 001102937700001 Biochemistry & Molecular Biology Biotechnology & Applied Microbiology Engineering, Chemical Biochemistry & Molecular Biology Biotechnology & Applied Microbiology Engineering Article Carbon dioxide fixation Ribulose-1 5-bisphosphate carboxylase/oxygenase Saccharomyces cerevisiae Delta-integration strategy CRISPR/Cas9