Identification of crucial roles of transcription factor IhfA on high production of free fatty acids in Escherichia coli
Transcription factor engineering has unique advantages in improving the performance of microbial cell factories due to the global regulation of gene transcription. Omics analyses and reverse engineering enable learning and subsequent incorporation of novel design strategies for further engineering....
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| Format: | Article |
| Language: | English |
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KeAi Communications Co., Ltd.
2024-03-01
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| Series: | Synthetic and Systems Biotechnology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405805X24000073 |
| _version_ | 1797272412716269568 |
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| author | Lixia Fang Ziyi Han Xueru Feng Xueyan Hao Mengxiao Liu Hao Song Yingxiu Cao |
| author_facet | Lixia Fang Ziyi Han Xueru Feng Xueyan Hao Mengxiao Liu Hao Song Yingxiu Cao |
| author_sort | Lixia Fang |
| collection | DOAJ |
| description | Transcription factor engineering has unique advantages in improving the performance of microbial cell factories due to the global regulation of gene transcription. Omics analyses and reverse engineering enable learning and subsequent incorporation of novel design strategies for further engineering. Here, we identify the role of the global regulator IhfA for overproduction of free fatty acids (FFAs) using CRISPRi-facilitated reverse engineering and cellular physiological characterization. From the differentially expressed genes in the ihfAL− strain, a total of 14 beneficial targets that enhance FFAs production by above 20 % are identified, which involve membrane function, oxidative stress, and others. For membrane-related genes, the engineered strains obtain lower cell surface hydrophobicity and increased average length of membrane lipid tails. For oxidative stress-related genes, the engineered strains present decreased reactive oxygen species (ROS) levels. These gene modulations enhance cellular robustness and save cellular resources, contributing to FFAs production. This study provides novel targets and strategies for engineering microbial cell factories with improved FFAs bioproduction. |
| first_indexed | 2024-03-07T14:28:58Z |
| format | Article |
| id | doaj.art-5654d1d4a8bb43939b3e13c6e67b15dc |
| institution | Directory Open Access Journal |
| issn | 2405-805X |
| language | English |
| last_indexed | 2024-03-07T14:28:58Z |
| publishDate | 2024-03-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Synthetic and Systems Biotechnology |
| spelling | doaj.art-5654d1d4a8bb43939b3e13c6e67b15dc2024-03-06T05:27:59ZengKeAi Communications Co., Ltd.Synthetic and Systems Biotechnology2405-805X2024-03-0191144151Identification of crucial roles of transcription factor IhfA on high production of free fatty acids in Escherichia coliLixia Fang0Ziyi Han1Xueru Feng2Xueyan Hao3Mengxiao Liu4Hao Song5Yingxiu Cao6Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China; Frontiers Research Institute for Synthetic Biology, Tianjin University, ChinaFrontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China; Frontiers Research Institute for Synthetic Biology, Tianjin University, ChinaFrontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China; Frontiers Research Institute for Synthetic Biology, Tianjin University, ChinaFrontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China; Frontiers Research Institute for Synthetic Biology, Tianjin University, ChinaFrontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China; Frontiers Research Institute for Synthetic Biology, Tianjin University, ChinaFrontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China; Frontiers Research Institute for Synthetic Biology, Tianjin University, ChinaFrontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China; Frontiers Research Institute for Synthetic Biology, Tianjin University, China; Corresponding author. Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China.Transcription factor engineering has unique advantages in improving the performance of microbial cell factories due to the global regulation of gene transcription. Omics analyses and reverse engineering enable learning and subsequent incorporation of novel design strategies for further engineering. Here, we identify the role of the global regulator IhfA for overproduction of free fatty acids (FFAs) using CRISPRi-facilitated reverse engineering and cellular physiological characterization. From the differentially expressed genes in the ihfAL− strain, a total of 14 beneficial targets that enhance FFAs production by above 20 % are identified, which involve membrane function, oxidative stress, and others. For membrane-related genes, the engineered strains obtain lower cell surface hydrophobicity and increased average length of membrane lipid tails. For oxidative stress-related genes, the engineered strains present decreased reactive oxygen species (ROS) levels. These gene modulations enhance cellular robustness and save cellular resources, contributing to FFAs production. This study provides novel targets and strategies for engineering microbial cell factories with improved FFAs bioproduction.http://www.sciencedirect.com/science/article/pii/S2405805X24000073Free fatty acidsCRISPRiihfACell membraneROS |
| spellingShingle | Lixia Fang Ziyi Han Xueru Feng Xueyan Hao Mengxiao Liu Hao Song Yingxiu Cao Identification of crucial roles of transcription factor IhfA on high production of free fatty acids in Escherichia coli Synthetic and Systems Biotechnology Free fatty acids CRISPRi ihfA Cell membrane ROS |
| title | Identification of crucial roles of transcription factor IhfA on high production of free fatty acids in Escherichia coli |
| title_full | Identification of crucial roles of transcription factor IhfA on high production of free fatty acids in Escherichia coli |
| title_fullStr | Identification of crucial roles of transcription factor IhfA on high production of free fatty acids in Escherichia coli |
| title_full_unstemmed | Identification of crucial roles of transcription factor IhfA on high production of free fatty acids in Escherichia coli |
| title_short | Identification of crucial roles of transcription factor IhfA on high production of free fatty acids in Escherichia coli |
| title_sort | identification of crucial roles of transcription factor ihfa on high production of free fatty acids in escherichia coli |
| topic | Free fatty acids CRISPRi ihfA Cell membrane ROS |
| url | http://www.sciencedirect.com/science/article/pii/S2405805X24000073 |
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