title: Fed-batch production of recombinant human calcitonin precursor fusion protein using Staphylococcus carnosus as an expression-secretion system.
abstract: A pH-auxostatic fed-batch process was developed for the secretory production of a fusion protein consisting of the pro-part of Staphylococcus hyicus lipase and two synthetic human calcitonin (hCT) precursor repeats under the control of a xylose-inducible promotor from Staphylococcus xylosus. Using glycerol as the energy source and pH-controlled addition of yeast extract resulted in the production of 2000 mg 1(-1) of the fusion protein (420 mg 1(-1) of the recombinant hCT precursor) within 14 h, reaching 45 g 1(-1) cell dry mass with Staphylococcus carnosus in a stirred-tank reactor. Product titer and space-time yield (30 mg calcitonin precursor 1(-1) h(-1)) were thus improved by a factor of 2, and 4.5, respectively, compared to Escherichia coli expression-secretion systems for the production of calcitonin precursors. Two hundred grams of the fusion protein was secreted by the recombinant S. carnosus on a 150-1 scale (scale-up factor of 50) with a minimum use of technical-grade yeast extract (40 mg fusion protein g(-1) yeast extract).
authors.affiliation: Lehrstuhl für Bioverfahrenstechnik, Technische Universität München, Garching, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Dilsen
authors.forename: W
authors.initials: W
authors.lastname: Paul
authors.forename: A
authors.initials: A
authors.lastname: Sandgathe
authors.forename: D
authors.initials: D
authors.lastname: Tippe
authors.forename: R
authors.initials: R
authors.lastname: Freudl
authors.forename: J
authors.initials: J
authors.lastname: Thömmes
authors.forename: M R
authors.initials: MR
authors.lastname: Kula
authors.forename: R
authors.initials: R
authors.lastname: Takors
authors.forename: C
authors.initials: C
authors.lastname: Wandrey
authors.forename: D
authors.initials: D
authors.lastname: Weuster-Botz
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D019149:Bioreactors; D002116:Calcitonin / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D003470:Culture Media; D005285:Fermentation; D005990:Glycerol / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000235:genetics; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000378:metabolism; D013210:Staphylococcus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
chemical_list: D000596:Amino Acids; D003470:Culture Media; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D002116:Calcitonin; D008049:Lipase; D005990:Glycerol