Engineering Halomonas bluephagenesis via small regulatory RNAs |
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| Institution: | 1. College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, China;2. MOE Key Lab of Bioinformatics, School of Life Sciences, Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, 100084, China;3. Center for Synthetic and Systems Biology, Tsinghua University, Beijing, 100084, China;4. MOE Key Laboratory for Industrial Biocatalysis, Dept Chemical Engineering, Tsinghua University, Beijing, 100084, China;5. Shandong Provincial Research Center for Bioinformatic Engineering and Technology, School of Life Sciences, Shandong University of Technology, Zibo, 255049, China;1. Department of Chemical and Biomolecular Engineering, The University of Tennessee, Knoxville, TN, USA;2. Center of Bioenergy Innovation, Oak Ridge National Laboratory, Oak Ridge, TN, USA;3. Chemical Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, 37830, USA;4. Department of Biological Engineering, Konkuk University, Seoul, South Korea;1. Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis, Saint Louis, MO, 63130, USA;2. The Edison Family Center for Genome Sciences and Systems Biology, Washington University School of Medicine in St. Louis, Saint Louis, MO, 63110, USA;3. Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, Saint Louis, MO, 63110, USA;4. Mass Spectrometry Resource, Division of Endocrinology, Metabolism, and Lipid Research, Department of Medicine, Washington University School of Medicine, St. Louis, MO, 63110, USA;5. Department of Molecular Microbiology, Washington University School of Medicine in St. Louis, Saint Louis, MO, 63110, USA;6. Department of Biomedical Engineering, Washington University in St. Louis, Saint Louis, MO, 63130, USA;7. Institute of Materials Science & Engineering, Washington University in St. Louis, Saint Louis, MO, 63130, USA;1. Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH, 4058, Basel, Switzerland;2. SIB Swiss Institute of Bioinformatics, Mattenstrasse 26, CH, 4058, Basel, Switzerland;3. Faculty of Science, University of Basel, Mattenstrasse 26, CH, 4058, Basel, Switzerland;1. National Engineering Research Center of Cereal Fermentation and Food Biomanufacturing, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China;2. Jiangsu Provincial Research Center for Bioactive Product Processing Technology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China |
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| Abstract: | Halomonas bluephagenesis, a robust and contamination-resistant microorganism has been developed as a chassis for “Next Generation Industrial Biotechnology”. The non-model H. bluephagenesis requires efficient tools to fine-tune its metabolic fluxes for enhanced production phenotypes. Here we report a highly efficient gene expression regulation system (PrrF1-2-HfqPa) in H. bluephagenesis, small regulatory RNA (sRNA) PrrF1 scaffold from Pseudomonas aeruginosa and a target-binding sequence that downregulate gene expression, and its cognate P. aeruginosa Hfq (HfqPa), recruited by the scaffold to facilitate the hybridization of sRNA and the target mRNA. The PrrF1-2-HfqPa system targeting prpC in H. bluephagenesis helps increase 3-hydroxyvalerate fraction in poly(3-hydroxybutyrate-co-3-hydroxyvalerate) to 21 mol% compared to 3.1 mol% of the control. This sRNA system repressed phaP1 and minD simultaneously, resulting in large polyhydroxybutyrate granules. Further, an sRNA library targeting 30 genes was employed for large-scale target identification to increase mevalonate production. This work expands the study on using an sRNA system not based on Escherichia coli MicC/SgrS-Hfq to repress gene expression, providing a framework to exploit new powerful genome engineering tools based on other sRNAs. |
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| Keywords: | Synthetic sRNA PHB PHA Mevalonate Metabolic engineering |
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