期刊
ACS SYNTHETIC BIOLOGY
卷 8, 期 4, 页码 633-640出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.8b00487
关键词
riboswitch; inducible and repressible systems; thermophiles; Clostridium thermocellum; gene regulation
资金
- National Renewable Energy Laboratory Director's Fellowship Program - Laboratory Directed Research and Development [06271601]
- US Department of Energy (DOE) Energy Efficiency and Renewable Energy (EERE) Fuel Cell Technologies Office [DE AC36 08 GO28308]
- National Renewable Energy Laboratory Research Data Initiative
Thermophilic bacteria are attractive hosts to produce bio-based chemicals. While various genetic manipulations have been employed in the metabolic engineering of thermophiles, a robust means to regulate gene expression in these bacteria (similar to 55 degrees C) is missing. Our bioinformatic search for various riboswitches in thermophilic bacteria revealed that major classes of riboswitches are present, suggesting riboswitches' regulatory roles in these bacteria. By building synthetic constructs incorporating natural and engineered purine riboswitch sequences originated from foreign species, we quantified respective riboswitches activities in repressing and up-regulating gene expression in Geobacillus thermoglucosidasius using a green fluorescence protein. The elicited regulatory response was ligand-concentration-dependent. We further demonstrated that riboswitch-mediated gene expression of adhE (responsible for ethanol production) in Clostridium thermocellum can modulate ethanol production, redirect metabolites, and control cell growth in the adhE knockout mutant. This work has made tunable gene expression feasible across different thermophiles for broad applications including biofuels production and gene-to-trait mapping.
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