title: Studies on the production of human parathyroid hormone by recombinant Escherichia coli.
abstract: Expression of human parathyroid hormone, hPTH(-1-84), by Escherichia coli N4830:pEX-PPTH was studied in controlled bioreactors. The hPTH is expressed as a fusion protein under control of the bacteriophage lambda PR promoter. In batch runs, low biomass concentrations but high specific hPTH productivities were obtained with complex TY (bactotryptone and yeast extract) medium whereas high biomass concentration and low specific productivities were found when fructose was used instead of bactotryptone (YF medium). The preinduction temperature was always 30 degrees C; the temperature shift to induce production of fusion protein was varied from 36 to 42 degrees C. Formation of hPTH passed a pronounced maximum as a function of induction temperature when using YF medium. However, the optimum temperature shift was 38 degrees C for both media used. For this temperature increase both media yielded about the same volumetric hPTH productivity (approx. 30 mg hPTH/1 per hour). By applying a fed-batch strategy for the YF medium, the productivity of the recombinant protein could be further increased more than fourfold. Compared to shake-flask experiments, the hPTH yield could be increased by a factor larger than 20.
authors.affiliation: GBF--Gesellschaft für Biotechnologische Forschung mbH, Biochemical Engineering, Braunschweig, Germany.
authors.forename: M P
authors.initials: MP
authors.lastname: Harder
authors.forename: E A
authors.initials: EA
authors.lastname: Sanders
authors.forename: E
authors.initials: E
authors.lastname: Wingender
authors.forename: W D
authors.initials: WD
authors.lastname: Deckwer
mesh_terms: D001709:Biotechnology; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics*; D005285:Fermentation; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D010281:Parathyroid Hormone / Q000096:biosynthesis* / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic; D013696:Temperature
chemical_list: D003470:Culture Media; D010281:Parathyroid Hormone; D011993:Recombinant Fusion Proteins