Purification and characterization of hyaluronic acid produced by Streptococcus zooepidemicus strain 3523-7
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Date
2013-09-17
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Publisher
Plovdiv University Press “Paisii Hilendarski”
Abstract
Hyaluronic acid (HA) is a hydrated gel and comprises repeating units of glucuronic acid and N-acetylglucosamine. Production and recovery of HA has gained great importance due to its vast clinical applications. In pursuit of obtaining highly pure HA, we have developed a fed-batch fermentation process using Streptococcus zooepidemicus in a 25 L bioreactor that resulted in a maximum yield of 2.3 g/L HA. In addition, we have devised an efficient method for separation and recovery of hyaluronic acid from a highly viscous broth by treating with trichloroacetic acid (0.1%) and charcoal (1-2%), passing through filtration (0.45 μm) and ultrafiltration that resulted in recovery of 72.2% of clinical grade HA with molecular weight of 2.5×106 Da. We have also characterized our purified HA using FTIR and NMR spectroscopy. These studies revealed the similarity in both the FTIR spectrum as well as NMR spectrum of both reference standard and purified HA from S. zooepidemicus indicating that the reported process is more efficient in terms of better yield and high quality (99.2%).
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Keywords
Hyaluronic acid, fed-batch fermentation, purification, FTIR analysis, NMR analysis, Streptococcus zooepidemicus