Rare Codon Analysis in Rickettsia Affecting Recombinant Protein Expression in Escherichia coli

2020 ◽  
Vol 4 (1) ◽  
pp. 30-35
Author(s):  
Mahfoud Bakli ◽  
Raul Pascalau ◽  
Laura Smuleac

Abstract Rickettsia species are important emerging pathogens causing rickettsial diseases, which are important cause death worldwide. The number of recombinant proteins used for diagnostic and therapeutic applications has increased dramatically, which is important in determination of protein function, structure and antigensity. Although E. coli is widely used expression system, the codon bias can hamper protein expression due to the presence of rare codons in gene sequence coding protein of interest. Using bioinformatics tools, rare codon analysis of rickettsial genes was performed and compared to not expressed proteins in both R. prowazekii and R. rickettsii. A negative correlation between frequencies of rare codons in Rickettsia and success of rickettsial protein expression was observed. This study suggested a useful tool to improve rickettsial recombinant protein expression in E. coli.

2021 ◽  
Vol 12 ◽  
Author(s):  
Gema Lozano Terol ◽  
Julia Gallego-Jara ◽  
Rosa Alba Sola Martínez ◽  
Adrián Martínez Vivancos ◽  
Manuel Cánovas Díaz ◽  
...  

Recombinant protein production for medical, academic, or industrial applications is essential for our current life. Recombinant proteins are obtained mainly through microbial fermentation, with Escherichia coli being the host most used. In spite of that, some problems are associated with the production of recombinant proteins in E. coli, such as the formation of inclusion bodies, the metabolic burden, or the inefficient translocation/transport system of expressed proteins. Optimizing transcription of heterologous genes is essential to avoid these drawbacks and develop competitive biotechnological processes. Here, expression of YFP reporter protein is evaluated under the control of four promoters of different strength (PT7lac, Ptrc, Ptac, and PBAD) and two different replication origins (high copy number pMB1′ and low copy number p15A). In addition, the study has been carried out with the E. coli BL21 wt and the ackA mutant strain growing in a rich medium with glucose or glycerol as carbon sources. Results showed that metabolic burden associated with transcription and translation of foreign genes involves a decrease in recombinant protein expression. It is necessary to find a balance between plasmid copy number and promoter strength to maximize soluble recombinant protein expression. The results obtained represent an important advance on the most suitable expression system to improve both the quantity and quality of recombinant proteins in bioproduction engineering.


PLoS ONE ◽  
2018 ◽  
Vol 13 (7) ◽  
pp. e0198715 ◽  
Author(s):  
Matteo Rovere ◽  
Alex Edward Powers ◽  
Dushyant Shailesh Patel ◽  
Tim Bartels

Author(s):  
N. Hemamalini ◽  
S. Ezhilmathi ◽  
A. Angela Mercy

Escherichia coli is the most extensively used organism in recombinant protein production. It has several advantages including a very short life cycle, ease of genetic manipulation and the well-known cell biology etc. which makes E. coli as the perfect host for recombinant protein expression. Despite many advantages, E. coli also have few disadvantages such as coupled transcription and translation and lack of eukaryotic post-translational modifications. These challenges can be overcome by adopting several strategies such as, using different E. coli expression vectors, changing the gene sequence without altering the functional domain, modified E. coli strain usage, changing the culture parameters and co-expression with a molecular chaperone. In this review, we present the level of strategies used to enhance the recombinant protein expression and its stability in E. coli.


2008 ◽  
Vol 135 (1) ◽  
pp. 34-44 ◽  
Author(s):  
Karin Dürrschmid ◽  
Helga Reischer ◽  
Wolfgang Schmidt-Heck ◽  
Thomas Hrebicek ◽  
Reinhard Guthke ◽  
...  

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