title: Production of soluble recombinant proteins in Escherichia coli: effects of process conditions and chaperone co-expression on cell growth and production of xylanase.
abstract: In this study, effects of temperature, inducer concentration, time of induction and co-expression of molecular chaperones (GroEL-GroES and DnaKJE), on cell growth and solubilization of model protein, xylanases, were investigated. The yield of soluble xylanases increased with decreasing cultivation temperature and inducer level. In addition, co-expression of DnaKJE chaperone resulted in increased soluble xylanases though the time of induction of chaperone and target protein had a bearing on this yield. A combination of chaperone co-expression and partial induction resulted in ∼40% (in DnaKJE) and 33% (in GroEL-GroES) of total xylanase yield in soluble fraction. However, the conditions for maximum yield of soluble r-XynB and maximum % soluble expression of r-XynB were different. Higher expression of soluble xylanases in a scalable semi-synthetic medium showed potential of the process for soluble enzyme production.
authors.affiliation: CSIR - Institute of Microbial Technology, Sector 39-A, Chandigarh 160036, India.
authors.forename: Kamna
authors.initials: K
authors.lastname: Jhamb
authors.forename: Debendra K
authors.initials: DK
authors.lastname: Sahoo
mesh_terms: D001709:Biotechnology / Q000379:methods*; D043364:Endo-1,4-beta Xylanases / Q000096:biosynthesis*; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology / Q000254:growth & development* / Q000378:metabolism*; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D018832:Molecular Chaperones / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012995:Solubility; D013696:Temperature; D013997:Time Factors
chemical_list: D018832:Molecular Chaperones; D011994:Recombinant Proteins; D007544:Isopropyl Thiogalactoside; D043364:Endo-1,4-beta Xylanases