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

Sujeong Park; Bo Ram Park; Deokyeol Jeong; Jongbeom Park; Ja Kyong Ko; Soo Jung Kim; Jeong Sun Kim; Yong Su Jin; Soo Rin Kim

doi:10.1016/j.procbio.2023.10.013

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

Sujeong Park, Bo Ram Park, Deokyeol Jeong, Jongbeom Park, Ja Kyong Ko, Soo Jung Kim, Jeong Sun Kim, Yong Su Jin, Soo Rin Kim

Research output: Contribution to journal › Article › peer-review

Abstract

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.

Original language	English (US)
Pages (from-to)	286-293
Number of pages	8
Journal	Process Biochemistry
Volume	134
DOIs	https://doi.org/10.1016/j.procbio.2023.10.013
State	Published - Nov 2023

Keywords

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

ASJC Scopus subject areas

Bioengineering
Biochemistry
Applied Microbiology and Biotechnology

Online availability

10.1016/j.procbio.2023.10.013

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Cite this

@article{320315184e7a417596d2351b5b47ee75,

title = "Functional expression of RuBisCO reduces CO2 emission during fermentation by engineered Saccharomyces cerevisiae",

abstract = "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.",

keywords = "5-bisphosphate carboxylase/oxygenase, CRISPR/Cas9, Carbon dioxide fixation, Delta-integration strategy, Ribulose-1, Saccharomyces cerevisiae",

author = "Sujeong Park and Park, {Bo Ram} and Deokyeol Jeong and Jongbeom Park and Ko, {Ja Kyong} and Kim, {Soo Jung} and Kim, {Jeong Sun} and Jin, {Yong Su} and Kim, {Soo Rin}",

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year = "2023",

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language = "English (US)",

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AU - Park, Sujeong

AU - Park, Bo Ram

AU - Jeong, Deokyeol

AU - Park, Jongbeom

AU - Ko, Ja Kyong

AU - Kim, Soo Jung

AU - Kim, Jeong Sun

AU - Jin, Yong Su

AU - Kim, Soo Rin

PY - 2023/11

Y1 - 2023/11

N2 - 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.

AB - 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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