High-level soluble expression of hIGF-1 fusion protein in recombinant Escherichia coli

Zhang, Danping; Wei, Peilian; Fan, Limei; Lian, Jiazhang; Huang, Lei; Cai, Jin; Xu, Zhinan

doi:10.1016/j.procbio.2010.05.014

Cited by 13 publications

(5 citation statements)

References 23 publications

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“…For example, the effect of different engineered hosts including BL21(DE3) pLysS, BL21(DE3) and Rosetta on soluble expression of recombinant TNF-α was assessed by papaneophytou et al Their results showed lower yield of soluble TNF-α in Rosetta compared to the other two hosts 14 . The effective role of the engineered strains on solubility demonstrated here is also in line with Zhang et al's study which has showed the higher solubility of IGF1-thioredoxin fusion in Rosetta-gami (DE3) than that in Rosetta (DE3) and Bl21 (DE3) bacteria 15 . Herein, the maximum soluble amount of scFv was achieved in E. coli Origami(DE3) which is a type of mutant strain with mutation in thioredoxin reductase (trxB) and glutathione reductase (gor) genes.…”

Section: Discussionsupporting

confidence: 90%

Machine learning modeling for solubility prediction of recombinant antibody fragment in four different E. coli strains

Hashemi

Basafa

Behravan

2022

Sci Rep

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The solubility of proteins is usually a necessity for their functioning. Recently an emergence of machine learning approaches as trained alternatives to statistical models has been evidenced for empirical modeling and optimization. Here, soluble production of anti-EpCAM extracellular domain (EpEx) single chain variable fragment (scFv) antibody was modeled and optimized as a function of four literature based numerical factors (post-induction temperature, post-induction time, cell density of induction time, and inducer concentration) and one categorical variable using artificial neural network (ANN) and response surface methodology (RSM). Models were established by the CCD experimental data derived from 232 separate experiments. The concentration of soluble scFv reached 112.4 mg/L at the optimum condition and strain (induction at cell density 0.6 with 0.4 mM IPTG for 24 h at 23 °C in Origami). The predicted value obtained by ANN for the response (106.1 mg/L) was closer to the experimental result than that obtained by RSM (97.9 mg/L), which again confirmed a higher accuracy of ANN model. To the author’s knowledge this is the first report on comparison of ANN and RSM in statistical optimization of fermentation conditions of E.coli for the soluble production of recombinant scFv.

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Section: Discussionsupporting

confidence: 90%

Machine learning modeling for solubility prediction of recombinant antibody fragment in four different E. coli strains

Hashemi

Basafa

Behravan

2022

Sci Rep

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“…E. coli Rosetta-gami (Merck KGaA, Darmstadt, Germany) strains are Origami strains that harbor the pRARE plasmid and thereby combine the advantages of Origami strains (disulfide bond formation) with that of Rosetta (providing tRNAs for rare codons). Zhang et al [188] compared the performance of BL21, Rosetta, and Rosetta-gami for the production of human insulin-like growth factor 1 (hIGF-1) and observed that Rosetta-gami clearly outperformed the other two strains. Furthermore, from Table 3 it can be concluded that E. coli strains are mainly used for the production of small proteins.…”

Section: Escherichia Coli Strains Used In Rppmentioning

confidence: 99%

Increasing recombinant protein production in Escherichia coli through metabolic and genetic engineering

Waegeman¹,

Soetaert²

2011

J Ind Microbiol Biotechnol

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Different hosts have been used for recombinant protein production, ranging from simple bacteria, such as Escherichia coli and Bacillus subtilis, to more advanced eukaryotes as Saccharomyces cerevisiae and Pichia pastoris, to very complex insect and animal cells. All have their advantages and drawbacks and not one seems to be the perfect host for all purposes. In this review we compare the characteristics of all hosts used in commercial applications of recombinant protein production, both in the area of biopharmaceuticals and industrial enzymes. Although the bacterium E. coli remains a very often used organism, several drawbacks limit its possibility to be the first-choice host. Furthermore, we show what E. coli strains are typically used in high cell density cultivations and compare their genetic and physiological differences. In addition, we summarize the research efforts that have been done to improve yields of heterologous protein in E. coli, to reduce acetate formation, to secrete the recombinant protein into the periplasm or extracellular milieu, and to perform post-translational modifications. We conclude that great progress has been made in the incorporation of eukaryotic features into E. coli, which might allow the bacterium to regain its first-choice status, on the condition that these research efforts continue to gain momentum.

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“…122 To date, several expression systems, such as E. coli, 123 yeast, 124 cell-free systems, 125 transgenic plants, 126 and rabbits, 127 have been applied to produce IGF-1. 123 Among these expression systems, recombinant E. coli has been more beneficial rather than other hosts of easy handling and culture, and high yields make it the most popular host to produce IGF-1. 128 Ranjbari et al (2015) expressed 0.694 g/L IGF-1 internally as an inclusion body that highly burdened the successive purification.…”

Section: Cfps System For Insulin-like Growth Factor I Productionmentioning

confidence: 99%

Cell‐free protein synthesis system: A new frontier for sustainable biotechnology‐based products

Maharjan,

Park

2023

Biotech and App Biochem

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Cell‐free protein synthesis (CFPS) system is an innovative technology with a wide range of potential applications that could challenge current thinking and provide solutions to environmental and health issues. CFPS system has been demonstrated to be a successful way of producing biomolecules in a variety of applications, including the biomedical industry. Although there are still obstacles to overcome, its ease of use, versatility, and capacity for integration with other technologies open the door for it to continue serving as a vital instrument in synthetic biology research and industry. In this review, we mainly focus on the cell‐free based platform for various product productions. Moreover, the challenges in the bio‐therapeutic aspect using cell‐free systems and their future prospective for the improvement and sustainability of the cell free systems.

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High-level soluble expression of hIGF-1 fusion protein in recombinant Escherichia coli

Cited by 13 publications

References 23 publications

Machine learning modeling for solubility prediction of recombinant antibody fragment in four different E. coli strains

Machine learning modeling for solubility prediction of recombinant antibody fragment in four different E. coli strains

Increasing recombinant protein production in Escherichia coli through metabolic and genetic engineering

Cell‐free protein synthesis system: A new frontier for sustainable biotechnology‐based products

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