Codon optimization enhances protein expression of human peptide deformylase in E. coli

Han, Jiwon; Choi, Yun‐Seok; Kim, Won-Je; Jeon, Young Ho; Lee, Seung Kyu; Lee, Bong-Jin; Ryu, Kyoung‐Seok

doi:10.1016/j.pep.2009.10.005

Cited by 28 publications

(15 citation statements)

References 30 publications

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“…Therefore, foreign gene sequences can impair the translational machinery of the host cells and elicit several stress responses that will contribute to the metabolic burden (Jana and Deb, 2005;Gustafsson et al, 2004;Kane, 1995;Schweder et al, 2002). Different strategies have been applied either by performing codon-optimization of the heterologous genes, according to the frequency of codon usage in the host (Burgess-Brown et al, 2008;Han et al, 2010;Hale and Thompson, 1998;Yang et al, 2004;Angov et al, 2008;Niemitalo et al, 2005), or through the co-expression of genes that encode the tRNAs that are scarce in the host (Calderone et al, 1996;Spanjaard et al, 1990;Saxena and Walker, 1992;Del et al, 1995). However, such approaches have shown some adverse effects, such as the lower specific activities of the target products due to unpredictable structural alterations (Gustafsson et al, 2004).…”

Section: Genetic Modificationsmentioning

confidence: 99%

Metabolic responses to recombinant bioprocesses in Escherichia coli

Carneiro

Ferreira

Rocha

2013

Journal of Biotechnology

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Escherichia coli has been widely used for the production of recombinant proteins. However, the unbalances between host metabolism and recombinant biosynthesis continue to hamper the efficiency of these recombinant bioprocesses. The additional drainage of biosynthetic precursors toward recombinant processes burdens severely the metabolism of cells that, ultimately, elicits a series of stress responses, reducing biomass growth and recombinant protein production. Several strategies to overcome these metabolic limitations have been implemented; however, in most cases, improvements in recombinant protein expression were achieved at the expense of biomass growth arrest, which significantly hampers the efficiency of recombinant bioprocesses. With the advent of high throughput techniques and modelling approaches that provide a system-level understanding of the cellular systems, it is now expected that new advances in recombinant bioprocesses are achieved. By providing means to deal with these systems, our understanding on the metabolic behaviour of recombinant cells will advance and can be further explored to the design of suitable hosts and more efficient and cost-effective bioprocesses. Here, we review the major metabolic responses associated with recombinant processes and the engineering strategies relevant to overcome these stresses. Moreover, the advantages of applying systems levels engineering strategies to enhance recombinant protein production in E. coli cells are discussed and future perspectives on the advances of mathematical modelling approaches to study these systems are exposed.

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Section: Genetic Modificationsmentioning

confidence: 99%

Metabolic responses to recombinant bioprocesses in Escherichia coli

Carneiro

Ferreira

Rocha

2013

Journal of Biotechnology

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“…This could be as a result of changes in secondary structure of refolded rNAM compared to the native form that was shown by data obtained from circular dichroism analysis (manuscript under review). Different approaches are designed to eradicate the inclusion bodies including codon optimization strategy, fusion tags utilization [22], changing the expression host [9], and culture condition [26,14,29] such as lower growth temperature [24].…”

Section: Discussionmentioning

confidence: 99%

Design and Expression Optimization of a Chimeric Derivative of NetB, Alpha-Toxin and Metallopeptidase Proteins as a Subunit Vaccine Against Clostridium perfringens

Katalani¹,

Nematzadeh

Ahmadian

et al. 2019

vacres

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Avian necrotic enteritis (NE) is a multi-virulence disease caused by the bacterium Clostridium perfringens. Several toxins of this bacteria are components of different candidate vaccines, and have been considered as important factors in pathogenesis of NE. A fusion subunit protein composed of immunodominant segments of NetB, Alpha-toxin and metallopeptidase proteins (NAM) was designed. The high level production of NE subunit vaccine candidate is important for the in vitro and in vivo evaluation and later to decrease the production costs. Therefore, the Response Surface Methodology (RSM) was used for optimizing E. coli culture condition. To this end, induction conditions including cell optical density prior induction (OD600nm), IPTG concentration, post-induction temperature and time were modified. The statistical analysis revealed that all variables except IPTG had significant effects on production and solubility of rNAM. A 7.39 fold increase in production of soluble rNAM was achieved when the post induction temperature, IPTG concentration, the pre-induction OD and time were 19 ºC, 0.55 mM, 0.8 respectively in 8 h after induction. Our study indicated that the RSM method is a simple and superior strategy for protein expression improvement which is considered as a major limitation in production of vaccine candidate and other recombinant proteins using different hosts. Katalani1 et al Design and Expression Optimization of a Chimeric … * Recombinant NAM concentration yield (mg/L) obtained from densitometry analysis with different expression conditions of IPTG concentration, OD600, temperature and induction time using a central composite design for four variables.

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“…The Peptide Mass Fingerprinting (PMF) and bioinformatics analysis indicated that the 23 kDa protein was identical to the N-terminus part of the target protein. We inferred that the 23 kDa protein was a partial product resulting from premature termination [15,16]. …”

Section: Discussionmentioning

confidence: 99%

Expression, Purification and Identification of CtCVNH, a Novel Anti-HIV (Human Immunodeficiency Virus) Protein from Ceratopteris thalictroides

Sun

Wang

2013

IJMS

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CVN (cyanovirin-N) is an anti-HIV protein. CVNH (cyanovirin-N homology) represents its homology. In a previous study, we first reported the full-length sequences of the CVNH gene cloned from Ceratopteris thalictroides. Based on the finding, the coding sequence of CtCVNH was optimized in the study, and then a pET prokaryotic expression vector was constructed. The purification and identification of CtCVNH protein were investigated, as well. SDS-PAGE analysis indicated that a 31 kDa protein was overexpressed and mainly accumulated in the soluble fraction. Only a single protein was obtained after the Ni- nitrilotriacetic acid (NTA) affinity chromatography. The purified protein was identified to be the recombinant CtCVNH by both Western blot and peptide mass fingerprinting analysis.

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Codon optimization enhances protein expression of human peptide deformylase in E. coli

Cited by 28 publications

References 30 publications

Metabolic responses to recombinant bioprocesses in Escherichia coli

Metabolic responses to recombinant bioprocesses in Escherichia coli

Design and Expression Optimization of a Chimeric Derivative of NetB, Alpha-Toxin and Metallopeptidase Proteins as a Subunit Vaccine Against Clostridium perfringens

Expression, Purification and Identification of CtCVNH, a Novel Anti-HIV (Human Immunodeficiency Virus) Protein from Ceratopteris thalictroides

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