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Production of a recombinant, immunogenic protein, P64k, of Neisseria meningitidis in Escherichia coli in fed-batch fermenters
Authors:Raúl Espinosa  Evelyn Caballero  Alexis Musacchio  Ricardo Silva
Institution:(1) División de Vacunas, Centro de Ingeniería Genética y Biotecnología, P.O. 6162, Cubanacán, Playa, Ciudad Habana, 10 600, Cuba
Abstract:P64k is a Neisseria meningitidis high molecular weight protein present in meningococcal vaccine preparations. The lpdA gene, codifying for this protein, was cloned in Escherichia coli and the P64k protein was expressed in Escherichia coli K12 W3110 under the control of the tryptophan promoter. The recombinant bacteria were grown in batch or fed-batch cultures. P64k was expressed as an intracellular soluble form at about 40% of the total cellular protein. A final productivity of 215 mg l–1 h–1 and 11 g cell dry wt l–1 were obtained when the fed-batch culture conditions were optimised, compared to 30% of total protein, and a productivity of 76 mg l–1 h–1 and 5.1 g cell dry wt l–1 in batch cultivation.
Keywords:fed-batch culture  immunogenic protein  Neisseria meningitidis  outer membrane protein  vaccine
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