Co-expression of the small heat shock protein, Lo18, with β-glucosidase in Escherichia coli improves solubilization and reveals various associations with overproduced heterologous protein, GroEL/ES

Ronez, Florian; Desroche, Nicolas; Arbault, Patrice; Guzzo, Jean

doi:10.1007/s10529-012-0854-2

Cited by 7 publications

(2 citation statements)

References 9 publications

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“…It is now known that molecular chaperones assist the nascent polypeptide to fold properly during the process of protein synthesis and thus prevent protein aggregation. Few molecular chaperones are found to improve folding and solubilization of misfolded protein, while other chaperones are involved in prevention of protein aggregation [25][26][27]. The commonly used molecular chaperones in E. coli are GroEL, GroES, DnaK, DnaJ and Trigger factor ( Table 2).…”

Section: Plasmids Carrying Molecular Chaperones For Optimization Of Pmentioning

confidence: 99%

Plasmids for Optimizing Expression of Recombinant Proteins in E. coli

Al-Hejin¹,

Bora²,

Ahmed³

2019

Plasmid

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Plasmids are important vectors for the transfer of genetic material among microbes. The transfer of plasmids causes transmission of genes involved in pathogenesis and survival, to the host bacteria leading to their evolution and adaptation to diverse environmental conditions. A large number of plasmids of varying sizes have been discovered and isolated from various microorganisms. Plasmids are also valuable tools to genetically manipulate microbes for various purposes including production of recombinant proteins. Escherichia coli is the most preferred microbe for production of recombinant proteins, due to rapid growth rate, cost-effectiveness, high yield of the recombinant proteins and easy scale-up process. Several plasmids have been designed to optimize the expression of heterologous proteins in E. coli. In order to circumvent the issues of protein refolding, the codon usage in E. coli, the absence of post-translational modifications, such as glycosylation and low recovery of functionally active recombinant proteins, various plasmids have been designed and constructed. This chapter summarizes the recent technological advancements that have extended the use of the E. coli expression system to produce more complex proteins, including glycosylated recombinant proteins and therapeutic antibodies.

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Section: Plasmids Carrying Molecular Chaperones For Optimization Of Pmentioning

confidence: 99%

Plasmids for Optimizing Expression of Recombinant Proteins in E. coli

Al-Hejin¹,

Bora²,

Ahmed³

2019

Plasmid

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“…It is also prefered to increase the solubility of the proteins in E. coli by co-expression of proteins with molecular chaperones is a frequently used method. In E. coli, chaperone proteins such as DnaK, DnaJ, GrpE, GroEL, GroES and trigger factors are widely used alone or in combination [11][12][13][14][15]. GroEL-GroES and DnaK-DnaJ-GrpE, the most efficient chaperone combinations, have been reported to achieve protein refolding [16][17][18].…”

Section: Introductionmentioning

confidence: 99%

High-Level Production of MMLV Reverse Transcriptase Enzyme in Escherichia Coli

Kaplan

İmamoğlu

Gökçe

2021

International Journal of Advances in Engineering and Pure Sciences

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Reverse transcriptase (RT) of Moloney murine leukemia virus (MMLV) is the most widely used enzyme for cDNA synthesis and RNA amplification. In this study, we aimed to produce MMLV RT enzyme recombinantly due to its importance in molecular studies. In this context, the DNA fragment encoding the MMLV RT enzyme was cloned into pTOLT plasmid and expressed in E. coli BL21 (DE3) pLysE cells. Since the high-level expression of the protein caused the protein molecules to aggregate in the inclusion bodies, co-expression of MMLV RT and chaperone plasmids (pG-KJE8, pGro7, pKJE7, pGTf2, pTf16) was performed to obtain the MMLV RT protein in soluble form. Contrary to our expectations, because it could not be obtained in soluble form, the protein was recovered from the inclusion bodies using refolding process. Finally, the protein was purified by affinity chromatography and the activity of the protein was checked using RT-PCR technique.

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Optimization of Culture Parameters and Novel Strategies to Improve Protein Solubility

Arya

Sabir

Bora

et al. 2014

Methods in Molecular Biology

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The production of recombinant proteins, in soluble form in a prokaryotic expression system, still remains a challenge for the biotechnologist. Innovative strategies have been developed to improve protein solubility in various protein overexpressing hosts. In this chapter, we would focus on methods currently available and amenable to "desired modifications," such as (a) the use of molecular chaperones; (b) the optimization of culture conditions; (c) the reengineering of a variety of host strains and vectors with affinity tags; and (d) optimal promoter strengths. All these parameters are evaluated with the primary objective of increasing the solubilization of recombinant protein(s) during overexpression in Escherichia coli.

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Co-expression of the small heat shock protein, Lo18, with β-glucosidase in Escherichia coli improves solubilization and reveals various associations with overproduced heterologous protein, GroEL/ES

Cited by 7 publications

References 9 publications

Plasmids for Optimizing Expression of Recombinant Proteins in E. coli

Plasmids for Optimizing Expression of Recombinant Proteins in E. coli

High-Level Production of MMLV Reverse Transcriptase Enzyme in Escherichia Coli

Optimization of Culture Parameters and Novel Strategies to Improve Protein Solubility

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