title: An insight into fusion technology aiding efficient recombinant protein production for functional proteomics.
abstract: Advancements in peptide fusion technologies to maximize the protein production has taken a big leap to fulfill the demands of post-genomics era targeting elucidation of structure/function of the proteome and its therapeutic applications, by over-expression in heterologous expression systems. Despite being most preferred protein expression system armed with variety of cardinal fusion tags, expression of the functionally active recombinant protein in E. coli remains plagued. The present review critically analyses the aptness of well-characterized fusion tags utilized for over-expression of recombinant proteins with improved solubility and their compatibility with downstream purification procedures. The combinatorial tandem affinity strategies have shown to provide more versatile options. Solubility decreasing fusion tags have proved to facilitate the overproduction of antimicrobial peptides. Efficient removal of fusion tags prior to final usage is of utmost importance and has been summarized discussing the efficiency of various enzymatic and chemical methods of tag removal. Unfortunately, no single fusion tag works as a magic bullet to completely fulfill the requirements of protein expression and purification in active form. The information provided might help in selection and development of a successful protocol for efficient recombinant protein production for functional proteomics.
authors: Department of Botany, University of Allahabad, Allahabad, 211002, India.
authors: Dinesh K
authors: DK
authors: Yadav
authors: Amity Institute of Biotechnology, Amity University, Haryana, Amity Education Valley, Gurgaon, Manesar, 122413, India.
authors: Neelam
authors: N
authors: Yadav
authors: Department of Biochemistry, Sri Venkateshawara College, University of Delhi, New Delhi, India.
authors: Sarika
authors: S
authors: Yadav
authors: Research and Scientific Unit, College of Nursing & Allied Health Sciences, Jazan University, Jazan, 45142, Saudi Arabia.
authors: Shafiul
authors: S
authors: Haque
authors: Amity Institute of Microbial Technology, Amity University, Sector 125, Noida, 201313, Uttar Pradesh, India. Electronic address: ntuteja@amity.edu.
authors: Narendra
authors: N
authors: Tuteja
mesh_terms: D000818:Animals; D001709:Biotechnology; D019921:Biotinylation; D000939:Epitopes / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D023281:Genomics; D006801:Humans; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D015202:Protein Engineering; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D011549:Pseudomonas / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012995:Solubility
chemical_list: D000939:Epitopes; D010455:Peptides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins
keywords: Affinity tags; Fusion protein; Proteases; Recombinant protein; SUMO; Solubility enhancing tag; Tag removal; Tandem affinity purification