Objective The use and commercial value of hyaluronic acid (HA) as an important element in the pharmaceutical, biomedical, and cosmetics industry is because of its purity. Four recombinant strains of Corynebacterium glutamicum containing different genes were used to produce HA. Results The production parameters were measured and strain 183.2, with the highest amount of HA (2.15 mg/ml), was selected for further experiments. HA was precipitated by different ratios of ethanol-isopropanol at 4 degrees C and - 20 degrees C. Active charcoal (1%) was added to the solvent precipitation mixture at pH 5 and 10. Finally, to achieve more purity and separation, gel filtration chromatography was used. The best result was obtained using an ethanol-isopropanol ratio of 1:1 of at - 20 degrees C, followed by active charcoal treatment at the acidic pH, and three fractions of the chromatography with molecular weights of 27, 27-110, and < 27 KDa were more analyzed with electrophoresis and FTIR. Conclusions The present study described a simple, economical, and reproducible method resulting in a high yield for low-MW HA from C. glutamicum.