scholarly journals Bicistronic Design-Based Continuous and High-Level Membrane Protein Production in Escherichia coli

2019 ◽  
Vol 8 (7) ◽  
pp. 1685-1690 ◽  
Author(s):  
Nico J. Claassens ◽  
Max Finger-Bou ◽  
Bart Scholten ◽  
Frederieke Muis ◽  
Jonas J. de Groot ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Protein Production ◽  
High Level

Development of Escherichia coli Strains That Withstand Membrane Protein-Induced Toxicity and Achieve High-Level Recombinant Membrane Protein Production

2016 ◽  
Vol 6 (2) ◽  
pp. 284-300 ◽  
Author(s):  
Dimitra Gialama ◽  
Kalliopi Kostelidou ◽  
Myrsini Michou ◽  
Dafni Chrysanthi Delivoria ◽  
Fragiskos N. Kolisis ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Protein Production ◽  
High Level

Shaping Escherichia coli for recombinant membrane protein production

2018 ◽  
Vol 365 (15) ◽  
Author(s):  
Alexandros Karyolaimos ◽  
Henry Ampah-Korsah ◽  
Zhe Zhang ◽  
Jan-Willem de Gier
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Protein Production

scholarly journals Stress-Responsive Systems Set Specific Limits to the Overproduction of Membrane Proteins in Bacillus subtilis

2009 ◽  
Vol 75 (23) ◽  
pp. 7356-7364 ◽  
Author(s):  
Jessica C. Zweers ◽  
Thomas Wiegert ◽  
Jan Maarten van Dijl
Keyword(s):  
Bacillus Subtilis ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Drug Targets ◽  
Protein Production ◽  
Cell Envelope ◽  
Regulatory System ◽  
Interesting Candidate ◽  
High Level ◽  
And Function

ABSTRACT Essential membrane proteins are generally recognized as relevant potential drug targets due to their exposed localization in the cell envelope. Unfortunately, high-level production of membrane proteins for functional and structural analyses is often problematic. This is mainly due to their high overall hydrophobicity. To develop new concepts for membrane protein overproduction, we investigated whether the biogenesis of overproduced membrane proteins is affected by stress response-related proteolytic systems in the membrane. For this purpose, the well-established expression host Bacillus subtilis was used to overproduce eight essential membrane proteins from B. subtilis and Staphylococcus aureus. The results show that the σW regulon (responding to cell envelope perturbations) and the CssRS two-component regulatory system (responding to unfolded exported proteins) set critical limits to membrane protein production in large quantities. The identified sigW or cssRS mutant B. subtilis strains with significantly improved capacity for membrane protein production are interesting candidate expression hosts for fundamental research and biotechnological applications. Importantly, our results pinpoint the interdependent expression and function of membrane-associated proteases as key parameters in bacterial membrane protein production.

scholarly journals NlpI Facilitates Deposition of C4bp on Escherichia coli by Blocking Classical Complement-Mediated Killing, Which Results in High-Level Bacteremia

2012 ◽  
Vol 80 (10) ◽  
pp. 3669-3678 ◽  
Author(s):  
Yu-ting Tseng ◽  
Shainn-Wei Wang ◽  
Kwang Sik Kim ◽  
Ying-Hsiang Wang ◽  
Yufeng Yao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Neonatal Meningitis ◽  
Content Type ◽  
E Coli ◽  
Classical Complement Pathway ◽  
High Level ◽  
Classical Complement

ABSTRACTNeonatal meningitisEscherichia coli(NMEC) is the most common Gram-negative organism that is associated with neonatal meningitis, which usually develops as a result of hematogenous spread of the bacteria. There are two key pathogenesis processes for NMEC to penetrate into the brain, the essential step for the development ofE. colimeningitis: a high-level bacteremia and traversal of the blood-brain barrier (BBB). Our previous study has shown that the bacterial outer membrane protein NlpI contributes to NMEC binding to and invasion of brain microvascular endothelial cells, the major component cells of the BBB, suggesting a role for NlpI in NMEC crossing of the BBB. In this study, we showed that NlpI is involved in inducing a high level of bacteremia. In addition, NlpI contributed to the recruitment of the complement regulator C4bp to the surface of NMEC to evade serum killing, which is mediated by the classical complement pathway. NlpI may be involved in the interaction between C4bp and OmpA, which is an outer membrane protein that directly interacts with C4bp on the bacterial surface. The involvement of NlpI in two key pathogenesis processes of NMEC meningitis may make this bacterial factor a potential target for prevention and therapy ofE. colimeningitis.

Functional Requirements for DjlA- and RraA-Mediated Enhancement of Recombinant Membrane Protein Production in the Engineered Escherichia coli Strains SuptoxD and SuptoxR

2017 ◽  
Vol 429 (12) ◽  
pp. 1800-1816 ◽  
Author(s):  
Dimitra Gialama ◽  
Dafni Chrysanthi Delivoria ◽  
Myrsini Michou ◽  
Artemis Giannakopoulou ◽  
Georgios Skretas
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Protein Production ◽  
Functional Requirements ◽  
Engineered Escherichia Coli

scholarly journals Use of Amino Acids as Inducers for High-Level Protein Expression in the Single-Protein Production System

2010 ◽  
Vol 76 (18) ◽  
pp. 6063-6068 ◽  
Author(s):  
S. Thangminlal Vaiphei ◽  
Lili Mao ◽  
Tsutomu Shimazu ◽  
Jung-Ho Park ◽  
Masayori Inouye
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Protein Production ◽  
Target Protein ◽  
Complete Suppression ◽  
Tryptophan Residues ◽  
Single Protein ◽  
High Level ◽  
Isotopic Incorporation ◽  
Specific Mrna

ABSTRACT By taking advantage of MazF, an ACA codon-specific mRNA interferase, Escherichia coli cells can be converted into a bioreactor producing only a single protein of interest by using an ACA-less mRNA for the protein. In this single-protein production (SPP) system, we engineered MazF by replacing two tryptophan residues in positions 14 and 83 with Phe (W14F) and Leu (W83L), respectively. Upon the addition of an inducer (IPTG [isopropyl-β-d-thiogalactopyranoside]), the mutated MazF [MazF(ΔW)] can still be produced even in the absence of tryptophan in the medium by using a Trp auxotroph, while a target protein having Trp residues cannot be produced. However, at 3 h after the addition of IPTG, the addition of tryptophan to the medium exclusively induces production of the target protein at a high level. A similar SPP system was also constructed with the use of a His-less protein [MazF(ΔH)] and a His auxotroph. Using these dual-induction systems, isotopic enrichments of 13C, 15N, and 2H were highly improved by almost complete suppression of the production of the unlabeled target protein. In both systems, isotopic incorporation reached more than 98% labeling efficiency, significantly reducing the background attributable to the unlabeled target protein.

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