Fermentation processes for cultivating streptococci and purification processes for obtaining cps therefrom

Patent metadata

Field	Value
Publication number	US-2016137973-A1
Application number	US-201615003263-A
Country	US
Kind code	A1
Filing date	Jan 21, 2016
Priority date	Dec 20, 2007
Publication date	May 19, 2016
Grant date	—

Abstract

This invention is in the field of bacterial cultures and specifically relates to the optimization of culture conditions to improve the production of bacterial capsular polysaccharides from Streptococcus strains in fed batch culture and to novel purification methods suitable for production scale purification of bacterial capsular polysaccharides from Streptococcus strains resulting in higher levels of purity than previously obtained for production scale.

First claim

1 . A cultivating medium comprising a Streptococcus strain, a phosphate source, a carbon source, a vitamin source, and an amino acid source to grow Streptococcus , wherein said vitamin source consists of six or fewer vitamins selected from the following list of seven vitamins: biotin, niacinamide, calcium pantothenate, riboflavin, thiamine hydrochloride, pyridoxine hydrochloride and folic acid, wherein two of the vitamins have to be calcium pantothenate and niacinamide.

2 . The cultivating medium of claim 1 , wherein said Streptococcus is Streptococcus agalactiae.

3 . The cultivating medium in claim 1 , wherein said phosphate source consists of K 2 HPO 4 , KH 2 PO 4 , Na 2 HPO 4 .H2O, NaH 2 PO 4 .H2O, or NaCl.

4 . The cultivating medium of claim 1 , wherein said carbon source is glucose.

5 . The cultivating medium of claim 1 , wherein said vitamin source consists of six or fewer from the following list of seven vitamins: biotin, niacinamide, calcium pantothenate, riboflavin, thiamine hydrochloride, pyridoxine hydrochloride, and folic acid, wherein two have to be calcium pantothenate and niacinamide.

6 . The cultivating medium of claim 1 , wherein said vitamin source consists of five or fewer from the following list of seven vitamins: biotin, niacinamide, calcium pantothenate, riboflavin, thiamine hydrochloride, pyridoxine hydrochloride, and folic acid, wherein two have to be calcium pantothenate and niacinamide.

7 . The cultivating medium of claim 1 , wherein said vitamin source consists of four or fewer from the following list of seven vitamins: biotin, niacinamide, calcium pantothenate, riboflavin, thiamine hydrochloride, pyridoxine hydrochloride, and folic acid, wherein two have to be calcium pantothenate and niacinamide.

8 . The cultivating medium of claim 1 , wherein said vitamin source consists of three or fewer from the following list of seven vitamins: biotin, niacinamide, calcium pantothenate, riboflavin, thiamine hydrochloride, pyridoxine hydrochloride, and folic acid, wherein two have to be calcium pantothenate and niacinamide.

9 . The cultivating medium of claim 1 , wherein said vitamin source consists of calcium pantothenate and niacinamide.

10 . The cultivating medium of claim 1 , wherein said amino acid source consists of nineteen or fewer from the following list of nineteen amino acids: alanine, arginine, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, aspartic acid, cysteine hydrochloride, glutamic acid, and tyrosine, wherein fifteen have to be arginine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, valine, cysteine hydrochloride, glutamic acid, and tyrosine.

11 . The cultivating medium of claim 1 , wherein said amino acid source consists of eighteen or fewer from the following list of nineteen amino acids: alanine, arginine, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, aspartic acid, cysteine hydrochloride, glutamic acid, and tyrosine, wherein fifteen have to be arginine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, valine, cysteine hydrochloride, glutamic acid, and tyrosine.

12 . The cultivating medium of claim 1 , wherein said amino acid source consists of seventeen or fewer from the following list of nineteen amino acids: alanine, arginine, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, aspartic acid, cysteine hydrochloride, glutamic acid, and tyrosine, wherein fifteen have to be arginine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, valine, cysteine hydrochloride, glutamic acid, and tyrosine.

13 . The cultivating medium of claim 1 , wherein said amino acid source consists of sixteen or fewer from the following list of nineteen amino acids: alanine, arginine, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, aspartic acid, cysteine hydrochloride, glutamic acid, and tyrosine, wherein fifteen have to be arginine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, valine, cysteine hydrochloride, glutamic acid, and tyrosine.

14 . The cultivating medium of claim 1 , wherein said amino acid source consists of arginine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, valine, cysteine hydrochloride, glutamic acid, and tyrosine.

15 . A cultivating medium comprising a Streptococcus strain, a yeast extract, a phosphate source, a carbon source, a vitamin source, and optionally an amino acid source to grow Streptococcus , wherein said vitamin source consists of four or fewer vitamins selected from the following list of five vitamins: biotin, niacinamide, riboflavin, thiamine hydrochloride and pyridoxine hydrochloride, wherein one of the vitamins has to be biotin.

16 . The cultivating medium of claim 15 , wherein said Streptococcus is Streptococcus agalactiae.

17 . The cultivating medium of claim 15 , wherein said vitamin source consists of four or fewer vitamins selected from the following list of five vitamins: biotin, niacinamide, riboflavin, thiamine hydrochloride and pyridoxine hydrochloride, wherein one of the vitamins has to be biotin.

18 . The cultivating medium of claim 15 , wherein said vitamin source consists of three or fewer vitamins selected from the following list of five vitamins: biotin, niacinamide, riboflavin, thiamine hydrochloride and pyridoxine hydrochloride, wherein one of the vitamins has to be biotin.

19 . The cultivating medium of claim 15 , wherein said vitamin source consists of two or fewer vitamins selected from the following list of five vitamins: biotin, niacinamide, riboflavin, thiamine hydrochloride and pyridoxine hydrochloride, wherein one of the vitamins has to be biotin.

20 . The cultivating medium of claim 15 , wherein said vitamin source consists of biotin.

Assignees

• Glaxosmithkline Biolog Sa

Inventors

• Costantino Paolo
• Norelli Francesco
• Berti Francesco
• Olivieri Roberto
• Bazzocchi Giulia
• Cicala Concetta
• Fontani Silvia

Classifications

• A61P31/04

  Antibacterial agents · CPC title

• C12P19/04

  Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds · CPC title

• C12N2500/34

  Sugars · CPC title

• C12Q3/00

  Condition responsive control processes (apparatus therefor C12M1/36; controlling or regulating in general G05) · CPC title

• C12N2500/32

  Amino acids · CPC title