title: Statistical approaches to maximize recombinant protein expression in Escherichia coli: a general review.
abstract: The supply of many valuable proteins that have potential clinical or industrial use is often limited by their low natural availability. With the modern advances in genomics, proteomics and bioinformatics, the number of proteins being produced using recombinant techniques is exponentially increasing and seems to guarantee an unlimited supply of recombinant proteins. The demand of recombinant proteins has increased as more applications in several fields become a commercial reality. Escherichia coli (E. coli) is the most widely used expression system for the production of recombinant proteins for structural and functional studies. However, producing soluble proteins in E. coli is still a major bottleneck for structural biology projects. One of the most challenging steps in any structural biology project is predicting which protein or protein fragment will express solubly and purify for crystallographic studies. The production of soluble and active proteins is influenced by several factors including expression host, fusion tag, induction temperature and time. Statistical designed experiments are gaining success in the production of recombinant protein because they provide information on variable interactions that escape the "one-factor-at-a-time" method. Here, we review the most important factors affecting the production of recombinant proteins in a soluble form. Moreover, we provide information about how the statistical design experiments can increase protein yield and purity as well as find conditions for crystal growth.
authors.affiliation: Veterinary School, University of Thessaly, Trikalon 224, Karditsa 43100, Greece; Institute for Research and Technology - Thessaly (I.RE.TE.TH.), The Centre for Research & Technology Hellas (CE.R.TH.), Technology Park of Thessaly, 1st Industrial Area, Volos 38500, Greece.
authors.forename: Christos P
authors.initials: CP
authors.lastname: Papaneophytou
authors.affiliation: Veterinary School, University of Thessaly, Trikalon 224, Karditsa 43100, Greece; Institute for Research and Technology - Thessaly (I.RE.TE.TH.), The Centre for Research & Technology Hellas (CE.R.TH.), Technology Park of Thessaly, 1st Industrial Area, Volos 38500, Greece. Electronic address: gkontopidis@vet.uth.gr.
authors.forename: George
authors.initials: G
authors.lastname: Kontopidis
mesh_terms: D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D023281:Genomics; D011499:Protein Processing, Post-Translational; D040901:Proteomics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins
keywords: Escherichia coli; Fractional factorial; Recombinant protein; Response surface methodology (RSM); Solubility enhancement; Statistically designed experiments