Abstract: A cassette sequence for the transformation of a host cell includes at least: a first nucleotide sequence encoding a peptide or protein of interest to be produced by the host cell. The first sequence is linked to a second nucleotide sequence providing resistance to a toxin or encoding an antitoxin peptide to the toxin. The nucleotide sequences are organized in such a way that production of the peptide encoded by the second nucleotide sequence(s) is translationally coupled to production of the peptide encoded by the first nucleotide sequence.

Abstract: Methods for the production of a bacterial toxin such as diphtheria toxin, and methods for producing conjugates of a bacterial toxins, are provided.

Abstract: Methods for producing a conjugate of a bacterial toxin, including diphtheria toxin or CRM197, are provided.

Abstract: Compositions and methods related to periplasmic expression of a toxin including diphtheria toxin or CRM197 are provided herein.

Abstract: A cassette sequence for the transformation of a host cell includes at least: a first nucleotide sequence encoding a peptide or protein of interest to be produced by the host cell. The first sequence is linked to a second nucleotide sequence providing resistance to a toxin or encoding an antitoxin peptide to the toxin. The nucleotide sequences are organized in such a way that production of the peptide encoded by the second nucleotide sequence(s) is translationally coupled to production of the peptide encoded by the first nucleotide sequence.

Abstract: Chemically defined media for the industrial-scale culture of Bordetella species.

Abstract: Compositions and methods related to periplasmic expression of a toxin including diphtheria toxin or CRM197 are provided herein.

Abstract: Methods for identifying culture conditions for a cell, the method comprising: (a) obtaining a genome scale stoichiometric metabolic model of the cell, and (b) performing constraint based optimization on the metabolic model of the cell using at least one yield based constraint.

Abstract: A process for fermenting Bordetella species comprising incubating a sample of bacteria of a Bordetella species in a first environment under at least one bvg (Bordetella virulence genes) modulating condition, for at least 5 generations, to produce a mature culture, and then incubating the mature culture in a second environment in the absence of the at least one bvg modulating condition.

Abstract: Polynucleotides are provided herein.

Abstract: Chemically defined media for the industrial-scale culture of Bordetella species.

Abstract: The present invention provides a process for periplasmic expression of a bacterial toxoid comprising the steps of: a) growing a culture of a gram negative host cell in a fermentation medium, wherein the host cell is transformed with a polynucleotide, and wherein the polynucleotide encodes the bacterial toxoid and a periplasmic signal sequence; or providing a gram negative host cell wherein the host cell is transformed with a polynucleotide, the polynucleotide encodes the bacterial toxoid and a periplasmic signal sequence and wherein the gram negative host cell comprises the bacterial toxoid expressed in the periplasm; a(i)) inducing expression of the bacterial toxoid; b) maturing the host cell, wherein the maturing step comprises: I) subjecting the host cell to a pH shock; II) incubating the host cell with no feed addition; and/or III) subjecting the host cell to a temperature below ?20° C.; and c) extracting the bacterial toxoid from the host cell wherein the extraction process comprises osmotic shock.

Abstract: A process for fermenting Bordetella species comprising incubating a sample of bacteria of a Bordetella species in a first environment under at least one bvg (Bordetella virulence genes) modulating condition, for at least 5 generations, to produce a mature culture, and then incubating the mature culture in a second environment in the absence of the at least one bvg modulating condition.

Abstract: The present invention provides a process for periplasmic expression of a bacterial toxoid comprising the steps of: a), growing a culture of a gram negative host cell in a fermentation medium, wherein the host cell is transformed with a polynucleotide, and wherein the polynucleotide encodes the bacterial toxoid and a periplasmic signal sequence; or providing a gram negative host cell wherein the host cell is transformed with a polynucleotide, the polynucleotide encodes the bacterial toxoid and a periplasmic signal sequence and wherein the gram negative host cell comprises the bacterial toxoid expressed in the periplasm; a(i)) inducing expression of the bacterial toxoid; b) maturing the host ceil, wherein the maturing step comprises; I) subjecting the host cell to a pH shock; II) incubating the host ceil with no feed addition; and/or III) subjecting the host ceil to a temperature below ?20° C.; and c) extracting the bacterial toxoid from the host cell wherein the extraction process comprises osmotic shock.

Abstract: Polynucleotides are provided herein.