Synthetic reconstruction of extreme high hydrostatic pressure resistance in Escherichia coli |
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Affiliation: | 1. State Key Laboratory of Ocean Engineering, Shanghai Jiao Tong University, Shanghai 200240, China;2. Deep-Sea Microbial Cell Biology, Department of Deep Sea Science, Sanya Institute of Deep-Sea Science and Engineering, Chinese Academy of Sciences, 62 Fenghuang Road, Sanya 572000, China;3. Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California, San Diego, La Jolla, CA, USA;4. State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai 200240, China |
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Abstract: | Although high hydrostatic pressure (HHP) is an interesting parameter to be applied in bioprocessing, its potential is currently limited by the lack of bacterial chassis capable of surviving and maintaining homeostasis under pressure. While several efforts have been made to genetically engineer microorganisms able to grow at sublethal pressures, there is little information for designing backgrounds that survive more extreme pressures. In this investigation, we analyzed the genome of an extreme HHP-resistant mutant of E. coli MG1655 (designated as DVL1), from which we identified four mutations (in the cra, cyaA, aceA and rpoD loci) causally linked to increased HHP resistance. Analysing the functional effect of these mutations we found that the coupled effect of downregulation of cAMP/CRP, Cra and the glyoxylate shunt activity, together with the upregulation of RpoH and RpoS activity, could mechanistically explain the increased HHP resistance of the mutant. Using combinations of three mutations, we could synthetically engineer E. coli strains able to comfortably survive pressures of 600–800 MPa, which could serve as genetic backgrounds for HHP-based biotechnological applications. |
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Keywords: | High hydrostatic pressure Engineering of stress resistance Central carbon metabolism RpoS activity Heat shock response |
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