4.6 Article

Functional expression of RuBisCO reduces CO2 emission during fermentation by engineered Saccharomyces cerevisiae

Journal

PROCESS BIOCHEMISTRY
Volume 134, Issue -, Pages 286-293

Publisher

ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2023.10.013

Keywords

Carbon dioxide fixation; Ribulose-1; 5-bisphosphate carboxylase/oxygenase; Saccharomyces cerevisiae; Delta-integration strategy; CRISPR/Cas9

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This study developed a mixotrophic CO2-fixing Saccharomyces cerevisiae by constructing a CO2-fixation pathway and increasing the gene copy number of ribulose-1,5-bisphosphate carboxylase-oxygenase. The resulting yeast strain exhibited increased ethanol concentration and yield and reduced CO2 emissions during xylose fermentation.
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.

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