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

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.

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.