Chemically defined medium for the industrial scale culture of a species of Bordetella

Patent metadata

Field	Value
Publication number	US-9765294-B2
Application number	US-201415021345-A
Country	US
Kind code	B2
Filing date	Sep 11, 2014
Priority date	Sep 13, 2013
Publication date	Sep 19, 2017
Grant date	Sep 19, 2017

Abstract

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

First claim

What is claimed is:

1. A chemically defined medium for the industrial scale culture of a species of Bordetella comprising:
(i) an iron component selected from the group consisting of Fe(II) complexed to an organic compound and Fe(III) complexed to an organic compound, where the organic compound is selected from heme, haemoglobin, myoglobin, transferrin, ferritin, lactoferrin, enterobactin, aerobactin, alcaligin, coprogen, ferrichrome, desferrioxamine, ferroxamine, hydroxamate, citrate and dihydroxybenzoylserine;
(ii) 3-(N-morpholino)propanesulfonic acid (MOPS);
(iii) dimethyl-β-cyclodextrin; and
(iv) an amino acid selected from the group consisting of aspartate at a concentration 1000 μM or greater, glycine at a concentration of 1000 μM or greater, methionine at a concentration of 500 μM or greater and leucine at a concentration of 1500 μM or greater,

wherein said chemically defined medium does not comprise FeSO4 or tris(hydroxymethyl)aminomethane.

2. A chemically defined medium according to claim 1 , further comprising a component selected from the group consisting of:
(i) 2 μM or greater copper;
(ii) 2 μM or greater magnesium;
(iii) an additive selected from the group consisting of zinc, cobalt, thiamine, riboflavin and pantothenate; and
(iv) an additive selected from the group consisting of 0.4 μM or greater biotin, 50 μM or greater calcium, 15 μM or greater niacin, and 25 μM or greater ascorbic acid.

3. A chemically defined medium according to claim 1 , further comprising an inorganic source of sulphur selected from the group consisting of thiosulfate, trithionate, tetrathionate, peroxodisulfate, sulphide and sulphite, and wherein said medium does not comprise an organic source of sulfur.

4. The chemically defined medium according to claim 3 comprising greater than 0.005 mM thiosulfate.

5. The chemically defined medium according to claim 3 comprising greater than 0.003 mM trithionate.

6. The chemically defined medium according to claim 3 comprising greater than 0.002 mM tetrathionate.

7. The chemically defined medium according to claim 3 comprising greater than 0.005 mM peroxodisulfate.

8. The chemically defined medium according to claim 3 comprising greater than 0.010 mM sulphide.

9. The chemically defined medium according to claim 3 comprising greater than 0.010 mM sulphite.

10. The chemically defined medium of claim 1 comprising MOPS at a concentration of greater than 2 mM.

11. The chemically defined medium according to claim 1 , further comprising copper in the form of copper chloride.

12. The chemically defined medium according to claim 1 , further comprising an inorganic source of nitrogen selected from an ammonium salt and ammonium chloride.

13. The chemically defined medium according to claim 1 , further comprising a source of carbon selected from the group consisting of glutamate, proline, citrate, lactate, acetate, pyruvate, fumarate and succinate.

14. The chemically defined medium according to claim 1 , further comprising a component selected from the group consisting of:
(i) greater than 0.1 μM zinc;
(ii) greater than 0.05 μM cobalt;
(iii) greater than 100 μM calcium;
(iv) greater than 20 μM niacin;
(v) greater than 50 μM ascorbic acid;
(vi) greater than 0.1 μM thiamine;
(vii) greater than 0.4 μM biotin;
(viii) greater than 0.1 μM riboflavin; and
(ix) greater than 0.1 μM pantothenate.

15. The chemically defined medium according to claim 1 further comprising an amino acid selected from the group consisting of:
(i) glutamate at a concentration of greater than 50 mM;
(ii) alanine at a concentration of greater than 1000 μM;
(iii) phenylalanine at a concentration of greater than 500 μ;
(iv) histidine at a concentration of greater than 50 μM;
(v) isoleucine at a concentration of greater than 500 μM;
(vi) lysine at a concentration of greater than 500 μM;
(vii) proline at a concentration of greater than 1000 μM;
(viii) serine at a concentration of greater than 500 μM;
(ix) valine at a concentration of greater than 1000 μM; and
(x) tyrosine at a concentration of greater than 25 μM.

16. The chemically defined medium according to claim 1 further comprising glutathione at a concentration of greater than 100 μM.

17. The chemically defined medium according to claim 1 further comprising a component selected from the group consisting of:
(i) chloride at a concentration of less than 45 mM;
(ii) acetate at a concentration of greater than 1 mM; and
(iii) potassium at a concentration of greater than 1 mM.

18. A fermentation process for growing a species of Bordetella in a chemically defined medium (CDM) comprising
(a) inoculating a chemically defined medium according to claim 1 with a species of Bordetella ; and
(b) maintaining the species of Bordetella in the chemically defined medium for a period of time sufficient to allow biomass accumulation.

Assignees

• Glaxosmithkline Biologicals Sa
• Glaxosmithkline Biologicals Sa

Inventors

• Dehottay Philippe Marc Helene
• Goffin Philippe
• Branco Dos Santos Filipe
• Teusink Bas

Classifications

• C12N1/20 Primary

  Bacteria; Culture media therefor · CPC title