title: High purity recombinant human growth hormone (rhGH) expression in Escherichia coli under phoA promoter.
abstract: ABSTACT Recombinant human Growth Hormone (rhGH) is an important protein for human growth and is in high demand in clinics. Hence, it is necessary to develop an efficient fermentation process to produce highly pure rhGH. In this study, rhGH was expressed in Escherichia coli under alkaline phosphatase (phoA) promoter. The cultivation conditions for high expression level and purity of rhGH were investigated. The best initial phosphate concentration for rhGH expression, out of the 4 levels of initial phosphate concentration tests performed, was 12.6 mmol/L. Subsequently, 2 fed-batch cultivations under low dissolved oxygen (DO) (0% - 10%) and high DO (20% - 30%) conditions were carried out. High purity rhGH (92%) was obtained from 20% - 30% DO-stat cultivation, although the biomass did not show any significant difference. In summary, this research provided an efficient fermentation process for high purity rhGH production from E. coli under phoA promoter, which can lower the production and purification costs for large-scale production of rhGH.
authors.affiliation: a State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology , Shanghai , People's Republic of China.
authors.forename: Hao
authors.initials: H
authors.lastname: Song
authors.affiliation: a State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology , Shanghai , People's Republic of China.
authors.forename: Jingxin
authors.initials: J
authors.lastname: Jiang
authors.affiliation: a State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology , Shanghai , People's Republic of China.
authors.forename: Xuedong
authors.initials: X
authors.lastname: Wang
authors.affiliation: b Institute of Food Science and Engineering, University of Shanghai for Science and Technology , Shanghai , People's Republic of China.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Zhang
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli; D029968:Escherichia coli Proteins / Q000235:genetics*; D005285:Fermentation; D019382:Human Growth Hormone / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D019382:Human Growth Hormone; D000469:Alkaline Phosphatase; C114089:phoA protein, E coli
keywords: Escherichia coli; alkaline phosphatase; fed-batch; fermentation strategy; rhGH; variant