Optimizing Recombinant Protein Production in the Escherichia coli Periplasm Alleviates Stress

摘要: In Escherichia coli, many recombinant proteins are produced in the periplasm. To direct these to this compartment, they equipped with an N-terminal signal sequence so that can traverse cytoplasmic membrane via protein-conducting Sec translocon. Recently, using single-chain variable antibody fragment BL1, we have shown harmonizing target gene expression intensity translocon capacity be used improve production yields of a protein Here, studied consequences improving BL1 periplasm by proteomics approach. When is not harmonized capacity, impaired translocation secretory proteins, misfolding/aggregation cytoplasm, and inefficient energy metabolism result poor growth low yields. The harmonization results normal growth, enhanced yields, and, surprisingly, composition proteome is-besides target-the same as cells empty vector. Thus, efficiently without causing any notable detrimental effects host. Finally, show under optimized conditions, small fraction released into extracellular milieu outer vesicles. We envisage our observations design strategies further E. coliIMPORTANCE bacterium coli widely produce proteins. Usually, trial-and-error-based screening approaches identify conditions lead high for optimization its accompanied alleviation stress. This indicates monitoring stress responses could facilitate