High-level expression of soluble recombinant proteins in Escherichia coli using an HE-maltotriose-binding protein fusion tag. 2018

Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China.

Recombinant proteins are commonly expressed in prokaryotic expression systems for large-scale production. The use of genetically engineered affinity and solubility enhancing fusion proteins has increased greatly in recent years, and there now exists a considerable repertoire of these that can be used to enhance the expression, stability, solubility, folding, and purification of their fusion partner. Here, a modified histidine tag (HE) used as an affinity tag was employed together with a truncated maltotriose-binding protein (MBP; consisting of residues 59-433) from Pyrococcus furiosus as a solubility enhancing tag accompanying a tobacco etch virus protease-recognition site for protein expression and purification in Escherichia coli. Various proteins tagged at the N-terminus with HE-MBP(Pyr) were expressed in E. coli BL21(DE3) cells to determine expression and solubility relative to those tagged with His6-MBP or His6-MBP(Pyr). Furthermore, four HE-MBP(Pyr)-fused proteins were purified by immobilized metal affinity chromatography to assess the affinity of HE with immobilized Ni2+. Our results showed that HE-MBP(Pyr) represents an attractive fusion protein allowing high levels of soluble expression and purification of recombinant protein in E. coli.

UI MeSH Term Description Entries
D009842 Oligopeptides Peptides composed of between two and twelve amino acids. Oligopeptide
D010450 Endopeptidases A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS. Endopeptidase,Peptide Peptidohydrolases
D010957 Plasmids Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS. Episomes,Episome,Plasmid
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D011993 Recombinant Fusion Proteins Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes. Fusion Proteins, Recombinant,Recombinant Chimeric Protein,Recombinant Fusion Protein,Recombinant Hybrid Protein,Chimeric Proteins, Recombinant,Hybrid Proteins, Recombinant,Recombinant Chimeric Proteins,Recombinant Hybrid Proteins,Chimeric Protein, Recombinant,Fusion Protein, Recombinant,Hybrid Protein, Recombinant,Protein, Recombinant Chimeric,Protein, Recombinant Fusion,Protein, Recombinant Hybrid,Proteins, Recombinant Chimeric,Proteins, Recombinant Fusion,Proteins, Recombinant Hybrid
D002846 Chromatography, Affinity A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D006639 Histidine An essential amino acid that is required for the production of HISTAMINE. Histidine, L-isomer,L-Histidine,Histidine, L isomer,L-isomer Histidine
D012995 Solubility The ability of a substance to be dissolved, i.e. to form a solution with another substance. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed) Solubilities

Related Publications

Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
August 1997, Protein expression and purification,
Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
July 2022, International journal of molecular sciences,
Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
February 2010, BMC biotechnology,
Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
January 2016, PloS one,
Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
May 1995, Journal of bacteriology,
Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
January 2007, Nature protocols,
Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
January 2012, Methods in molecular biology (Clifton, N.J.),
Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
January 1992, Chinese journal of biotechnology,
Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
April 2016, Protein expression and purification,
Yingqian Han, and Wanying Guo, and Bingqian Su, and Yujie Guo, and Jiang Wang, and Beibei Chu, and Guoyu Yang
November 2008, Protein expression and purification,
Copied contents to your clipboard!