Separation Science and Technology, Vol.44, No.4, 906-923, 2009
Purification and Characterization of Microbial Hyaluronic Acid by Solvent Precipitation and Size-Exclusion Chromatography
Samples of hyaluronic acid (HA) produced by submerged fermentation using a synthetic culture medium were recovered, purified, fractioned, and characterized using solvent precipitation and size-exclusion chromatography (SEC). The samples showed a wide molar mass distribution in the range 103-107 Da, most of which had an average molar mass between 104 and 105 Da after purification by sequential precipitations. Fractions of HA with molar mass above 105 Da were purified by SEC in a semi-preparative scale with nearly no protein contamination. Characterization and fractionation of the HA was carried out by SEC in a analytical and semi-preparative scale using Shodex OHPak SB806M HQ and Superose 6 columns, respectively.
Keywords:Hyaluronic acid;precipitation;pullulan;recovery and purification;size-exclusion chromatography