Genetic engineering system for syngas-utilizing acetogen, Eubacterium limosum KIST612

Jiyeong Jeong; Ji Yeon Kim; Byeonghyeok Park; In Geol Choi; In Seop Chang

doi:10.1016/j.biteb.2020.100452

Genetic engineering system for syngas-utilizing acetogen, Eubacterium limosum KIST612

Jiyeong Jeong, Ji Yeon Kim, Byeonghyeok Park, In Geol Choi, In Seop Chang

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

24 Citations (Scopus)

Abstract

Acetogens are capable of fixing inorganic gases to organic acids with/without bio-alcohols via the Wood-Ljungdahl pathway. Because of their potential to produce value-added metabolites, acetogens have received attention in the field of biorefinery. Eubacterium limosum KIST612 is one of the promising acetogens, which has been well studied for its syngas utilizing ability, and its genomic data are available. A strain-specific genetic engineering system for maximal benefits has not yet been studied in the strain, though. In the present study, we first developed a foreign gene expression system using a native constitutive promoter and the CRISPR/Cas9-based gene editing system for KIST612. Transcriptomics-based candidate selection of native promoter and GUS reporter gene assay enabled the identification of promoter of rubredoxin oxidoreductase (P_rbo) as a strong constitutive promoter in the strain. Application of Cas9-mediated genome editing was able to effectively delete the pyrF gene in the chromosome of KIST612.

Original language	English
Article number	100452
Journal	Bioresource Technology Reports
Volume	11
DOIs	https://doi.org/10.1016/j.biteb.2020.100452
Publication status	Published - 2020 Sept

Bibliographical note

Funding Information:
This work was supported by the C1 Gas Refinery Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT ( NRF-2015M3D3A1A01064883 ) and New & Renewable Energy Core Technology Program of the Korea Institute of Energy Technology Evaluation and Planning (KETEP) grants from the Ministry of Trade, Industry and Energy (No. 20173010092460 ).

Publisher Copyright:
© 2020 Elsevier Ltd

Keywords

Cas9-mediated genome editing
Eubacterium limosum
Native constitutive promoter
Syngas-utilizing acetogen

ASJC Scopus subject areas

Bioengineering
Renewable Energy, Sustainability and the Environment
Environmental Engineering
Waste Management and Disposal

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.biteb.2020.100452

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title = "Genetic engineering system for syngas-utilizing acetogen, Eubacterium limosum KIST612",

abstract = "Acetogens are capable of fixing inorganic gases to organic acids with/without bio-alcohols via the Wood-Ljungdahl pathway. Because of their potential to produce value-added metabolites, acetogens have received attention in the field of biorefinery. Eubacterium limosum KIST612 is one of the promising acetogens, which has been well studied for its syngas utilizing ability, and its genomic data are available. A strain-specific genetic engineering system for maximal benefits has not yet been studied in the strain, though. In the present study, we first developed a foreign gene expression system using a native constitutive promoter and the CRISPR/Cas9-based gene editing system for KIST612. Transcriptomics-based candidate selection of native promoter and GUS reporter gene assay enabled the identification of promoter of rubredoxin oxidoreductase (Prbo) as a strong constitutive promoter in the strain. Application of Cas9-mediated genome editing was able to effectively delete the pyrF gene in the chromosome of KIST612.",

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note = "Funding Information: This work was supported by the C1 Gas Refinery Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT ( NRF-2015M3D3A1A01064883 ) and New & Renewable Energy Core Technology Program of the Korea Institute of Energy Technology Evaluation and Planning (KETEP) grants from the Ministry of Trade, Industry and Energy (No. 20173010092460 ). Publisher Copyright: {\textcopyright} 2020 Elsevier Ltd",

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AU - Chang, In Seop

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Genetic engineering system for syngas-utilizing acetogen, Eubacterium limosum KIST612

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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