title: [Optimization of expression conditions of an induction strategy for improving liver targeted interferon (IFN-CSP) production in E. coli].
abstract: Expression conditions of induction strategies for the cytoplasmic inclusion bodies (IBs) production of liver targeted interferon IFN-CSP by recombinant Escherichia coli (E. coli) BL21 (DE3) were optimized in shake-flask cultures in this study. The factors of the optimized protocol included in the present study were pH, inducer IPTG (isopropyl beta-D-thiogalactoside) concentration, culture growth temperature, incubation time and induction point. The effects of those factors were investigated by 'single variable at a time' method, aimed to analyze characterization of the recombinant strain. Orthogonal experimental design was further used to optimize the above critical factors for IFN-CSP production. According to the expression optimization result, it was confirmed that the main influence factors were cell density and induction temperature. The IFN-CSP gene expression optimized conditions were: pH value of the culture medium was 6.0, culture temperature 37 degrees C, adding IPTG to final concentration 0.4 mmol/L when the recombinant strain growth density OD600 achieved 0.8 and induction time 4 h. At this point, the IBs represented 74.3% of the total cellular protein. Compared with the non-optimized condition, IFN-CSP production obtained in optimized induction strategies were increased by approx. 1.2-fold. The optimized induction strategy yielded 688.8 mg/L of IFN-CSP, providing experimental data to study the biology activity and productive technology of IFN-CSP.
authors.forename: Yanting
authors.initials: Y
authors.lastname: Huang
authors.forename: Xuemei
authors.initials: X
authors.lastname: Lu
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.forename: Xiaobao
authors.initials: X
authors.lastname: Jin
authors.forename: Jiayong
authors.initials: J
authors.lastname: Zhu
mesh_terms: D001709:Biotechnology / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism*; D015870:Gene Expression; D007372:Interferons / Q000096:biosynthesis*; D008099:Liver; D013696:Temperature
chemical_list: D003470:Culture Media; D007372:Interferons