Efficient process for purification of high molecular weight hyaluronic acid

A technology of hyaluronic acid and molecular weight, applied in the effective field for purifying high-molecular-weight hyaluronic acid, can solve problems such as environmental hazards, and achieve the effect of saving cost and reducing load

Inactive Publication Date: 2009-08-19
RELIANCE LIFE SCI PVT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of such solvents creates handling problems in scale-up operations and leads to environmental hazards

Method used

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  • Efficient process for purification of high molecular weight hyaluronic acid
  • Efficient process for purification of high molecular weight hyaluronic acid
  • Efficient process for purification of high molecular weight hyaluronic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0115] Example 1: Bacterial strains and media. S. equi subsp. zooepidemic ATCC 39920 was obtained from the American Type Culture Collection. Bacteria were maintained on brain heart infusion agar or tryptone soy broth.

Embodiment 2

[0116] Example 2: Estimation of HA

[0117] After precipitation with an equal volume of isopropanol and redissolution in 3% sodium acetate solution, it was determined by carbazole (Bitter, T., and Muir, M., A modified uronic acid carbazole reaction. Anal Biochem 4: 330-334, 1962 ) routinely estimates HA in fermentation broth.

Embodiment 3

[0118] Embodiment 3: Culture medium optimization

[0119] For the medium optimization experiment, after 2.5% casein hydrolyzate enzyme, 1% yeast extract, 0.2% K 2 HPO 4 , 0.15% NaCl, 0.04% MgSO 4 ·7H 2 Streptococcus zooepidemicus was cultured in a medium consisting of O and 2% carbon source (sucrose). The organisms were grown in Erlenmeyer flasks at 37°C for 24 hours at 200 rpm. To evaluate the effect of metal ions on HA production, at a final concentration of 0.025%, filter-sterilized CuSO 4 , ZnSO 4 and MnSO 4 The solution was added to the medium. see Figure 5 .

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Abstract

The present invention provides an efficient and inproved process for the production and purification of high molecular weight hyaluronic acid (HA). One embodiment relates to providing growth conditions for production of high molecular weight HA in Streptococcus zooepidemicus. For example, growth in medium comprising a lower protein concentration, e.g., 1% casein hydrolyzate, and a higher sugar concentration, e.g., 5% sucrose, gave 5-6 g / L of high molecular weight HA in the range of 3.5-4.0 x 10Da. The present invention also provides an efficient process for the purification of HA comprising treatment with silica gel then active carbon, and subsequent diafiltration with less volume of solvent.

Description

[0001] Cross references to related applications [0002] This application claims the benefit of Indian Provisional Application Nos. 1065 / MUM / 2006, filed 6th July, 2006 and 1874 / MUM / 2006, filed 13th November, 2006, the contents of which are hereby incorporated by reference in their entirety. Field of Invention [0003] The present invention relates to improved techniques for the production and purification of hyaluronic acid and its salts. The invention also relates to the production and process optimization and purification of hyaluronic acid and its salts with biomedical applications. Background of the Invention [0004] Hyaluronic acid (HA) is a naturally occurring biopolymer with biological functions in bacteria and higher animals including humans. Naturally occurring HA can be found in higher animal tissues, especially as a spacer filler (Balazs, Viscosupplementation: a new concept in the treatment of osteoarthritis. J. Rheumatol. Suppl.; 39: 3-9, (Aug. 1993)) . It is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/26C08B37/00
CPCC08B37/0003C08B37/0072C12P19/26
Inventor V·兰加斯瓦米V·桑托什J·达门德拉V·纳塔拉H·维兰卡A·卡帕特
Owner RELIANCE LIFE SCI PVT
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