scholarly journals Periplasmic Expression of 4/7 α-Conotoxin TxIA Analogs in E. coli Favors Ribbon Isomer Formation – Suggestion of a Binding Mode at the α7 nAChR

2019 ◽  
Vol 10 ◽  
Author(s):  
Yamina El Hamdaoui ◽  
Xiaosa Wu ◽  
Richard J. Clark ◽  
Julien Giribaldi ◽  
Raveendra Anangi ◽  
...  
Keyword(s):  
Binding Mode ◽  
E Coli ◽  
Periplasmic Expression ◽  
Α7 Nachr

Procaryotic Expression of Single-Chain Variable-Fragment (scFv) Antibodies: Secretion in L-Form Cells of Proteus mirabilis Leads to Active Product and Overcomes the Limitations of Periplasmic Expression in Escherichia coli

1998 ◽  
Vol 64 (12) ◽  
pp. 4862-4869 ◽  
Author(s):  
Jörg F. Rippmann ◽  
Michaela Klein ◽  
Christian Hoischen ◽  
Bodo Brocks ◽  
Wolfgang J. Rettig ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Proteus Mirabilis ◽  
Binding Activity ◽  
Host Cells ◽  
Heterologous Proteins ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
E Coli ◽  
Active Product ◽  
Periplasmic Expression

ABSTRACT Recently it has been demonstrated that L-form cells ofProteus mirabilis (L VI), which lack a periplasmic compartment, can be efficiently used in the production and secretion of heterologous proteins. In search of novel expression systems for recombinant antibodies, we compared levels of single-chain variable-fragment (scFv) production in Escherichia coliJM109 and P. mirabilis L VI, which express four distinct scFvs of potential clinical interest that show differences in levels of expression and in their tendencies to form aggregates upon periplasmic expression. Production of all analyzed scFvs in E. coli was limited by the severe toxic effect of the heterologous product as indicated by inhibition of culture growth and the formation of insoluble aggregates in the periplasmic space, limiting the yield of active product. In contrast, the L-form cells exhibited nearly unlimited growth under the tested production conditions for all scFvs examined. Moreover, expression experiments with P. mirabilis L VI led to scFv concentrations in the range of 40 to 200 mg per liter of culture medium (corresponding to volume yields 33- to 160-fold higher than those with E. coli JM109), depending on the expressed antibody. In a translocation inhibition experiment the secretion of the scFv constructs was shown to be an active transport coupled to the signal cleavage. We suppose that this direct release of the newly synthesized product into a large volume of the growth medium favors folding into the native active structure. The limited aggregation of scFv observed in the P. mirabilis L VI supernatant (occurring in a first-order-kinetics manner) was found to be due to intrinsic features of the scFv and not related to the expression process of the host cells. The P. mirabilis L VI supernatant was found to be advantageous for scFv purification. A two-step chromatography procedure led to homogeneous scFv with high antigen binding activity as revealed from binding experiments with eukaryotic cells.

scholarly journals Decoding Antibacterial and Antibiofilm Properties of Cinnamon and Cardamom Essential Oils: A Combined Molecular Docking and Experimental Study

2021 ◽  
Author(s):  
Elahe Pourkhosravani ◽  
fatemeh dehghan nayeri ◽  
Mitra Mohammadi Bazargani
Keyword(s):  
Cell Attachment ◽  
Antibacterial Properties ◽  
Binding Mode ◽  
Dilution Method ◽  
Antibiofilm Activity ◽  
Bacterial Strains ◽  
Biofilm Growth ◽  
Computational Docking ◽  
E Coli ◽  
Industrial Strains

Abstract This study sets out to compare the antibacterial and antibiofilm profiles of Ci/Ca EOs alone and in combination together against infectious bacterial strains. MIC assay was carried out to survey the effectiveness of prepared EOs by two-fold serial dilution method and MTT evaluation. Synergic antibacterial properties of EOs against target strains were studied by using checkerboard titration method. Biofilm growth and development were evaluated using CV and XTT reduction assays. Antibacterial activity was observed for EOs against both bacterial strains with stronger activity for CiEO against both bacteria. The synergistic antibacterial effect was observed only against B. subtilis. Based on the FIC index, combinations could not inhibit the growth of E. coli. The pure EOs and their combination inhibited cell attachment for both studied bacteria with stronger effect on E. coli. CV and XTT reduction assays results showed that Ci EO and its combination with CaEO had the highest antibiofilm activity at lowest MIC value 0.08% and 0.04/0.02% against biofilm formed by E. coli and B. subtilis respectively, indicating a high antibiofilm potential. Computational docking analyses also postulated that the active constituents of evaluated EOs have the potential to interact with different bacterial targets, suggested binding mode of action of EOs metabolites. By and large, synergistic anti-biofilm properties of EOs may provide further options for developing novel formula to inhibit a variety of infectious clinical and industrial strains without (or less) toxicity effects on human body.

scholarly journals 1,2H hyperfine spectroscopy and DFT modeling unveil the demethylmenasemiquinone binding mode to E. coli nitrate reductase A (NarGHI)

2020 ◽  
Vol 1861 (8) ◽  
pp. 148203
Author(s):  
Maryam Seif Eddine ◽  
Frédéric Biaso ◽  
Julia Rendon ◽  
Eric Pilet ◽  
Bruno Guigliarelli ◽  
...  
Keyword(s):  
Nitrate Reductase ◽  
Binding Mode ◽  
E Coli ◽  
Dft Modeling ◽  
Hyperfine Spectroscopy

scholarly journals New insights into the binding mode of pyridine-3-carboxamide inhibitors of E. coli DNA gyrase

2019 ◽  
Vol 27 (16) ◽  
pp. 3546-3550 ◽  
Author(s):  
Sarah Narramore ◽  
Clare E.M. Stevenson ◽  
Anthony Maxwell ◽  
David M. Lawson ◽  
Colin W.G. Fishwick
Keyword(s):  
Dna Gyrase ◽  
Binding Mode ◽  
E Coli

Effect of DnaK/DnaJ/GrpE and DsbC Chaperons on Periplasmic Expression of Fab Antibody by E. coli SEC Pathway

2017 ◽  
Vol 25 (1) ◽  
pp. 67-74 ◽  
Author(s):  
Hassan Dariushnejad ◽  
Safar Farajnia ◽  
Nosratollah Zarghami ◽  
Maryam Aria ◽  
Asghar Tanomand
Keyword(s):  
E Coli ◽  
Periplasmic Expression ◽  
Sec Pathway

Crystal structure of E. coli ZinT with one zinc-binding mode and complexed with citrate

2018 ◽  
Vol 500 (2) ◽  
pp. 139-144 ◽  
Author(s):  
Jinli Chen ◽  
Lulu Wang ◽  
Fei Shang ◽  
Yuesheng Dong ◽  
Nam-Chul Ha ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Binding Mode ◽  
Zinc Binding ◽  
E Coli

scholarly journals Molecular Modeling Approach for the Discovery of Escherichia coli O157:H7 Interaction Inhibitors of Natural Product Compounds

2019 ◽  
Vol 31 (10) ◽  
pp. 2287-2290
Author(s):  
S. Pitchuanchom ◽  
M. Nontakitticharoen ◽  
H. Thaisuchat
Keyword(s):  
Escherichia Coli ◽  
Binding Mode ◽  
Escherichia Coli O157 ◽  
Structure Based Drug Design ◽  
E Coli ◽  
Speed Up ◽  
Automated Docking ◽  
Binding Orientations ◽  
Natural Product Compounds ◽  
Novel Drug

The aim of this study is to report the development of Escherichia coli O157:H7 template for structure-based drug design. This template was validated by redocking with crystal ligand I. The results showed a good matching of docked and the crystallographic binding orientations with root mean square deviation (RMSD) less than 2.0 Å. Moreover, the developed template was applied to predict binding mode of 19 known E. coli inhibitors and 40 natural products. The results showed that the binding energy of almost E. coli inhibitors was related to their biological activity. The use of developed E. coli O157:H7 template in automated docking could speed up the process of novel drug discovery by allowing designed inhibitors to be tested computationally before the compounds are synthesized.

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