Preparation, purification, and characterization of low-molecular-weight hyaluronic acid
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ORIGINAL RESEARCH PAPER
Preparation, purification, and characterization of low-molecular-weight hyaluronic acid Mohammad Karami . Mahvash Khodabandeh Shahraky Fatemeh Tabandeh . Dina Morshedi . Saeed Aminzade
. Masume Ranjbar .
Received: 16 June 2020 / Accepted: 22 October 2020 Ó Springer Nature B.V. 2020
Abstract 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 °C and - 20 °C. Active charcoal (1%) was added to the solvent precipitation mixture at pH 5 and 10.
M. Karami M. K. Shahraky (&) M. Ranjbar F. Tabandeh D. Morshedi S. Aminzade Department of Bioprocess Engineering, National Institute of Genetic Engineering and Biotechnology (NIGEB), Sharake–Pajoohesh, Km 15, Tehran-Karaj Highway, P.O.Box: 14965/161, Tehran, Iran e-mail: [email protected] M. Karami e-mail: [email protected] M. Ranjbar e-mail: [email protected] F. Tabandeh e-mail: [email protected] D. Morshedi e-mail: [email protected] S. Aminzade e-mail: [email protected]
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 °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. Keywords Low-MW HA Purification Corynebacterium glutamicum Active charcoal Gel filtration chromatography
Introduction Hyaluronic acid is a natural polymer with high viscosity and viscoelastic properties and many applications in the pharmaceutical and cosmetic industries (Sze et al. 2016; Heldin et al. 2019), and its production and purification are, therefore, critical. The difficulties and limitations of hyaluronic acid extraction from animal sources have motivated its production by microbial fermentation, but its production from a microbial source such as Streptococcus zooepidemicus is also subject to many obstacles due to its pathogenicity and endotoxins in wild-type strains. Therefore, the
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Biotechnol Lett
production of hyaluronic acid from non-pathogenic bacteria has become a common practice (Lu et al. 2016; Pan et al. 2017). Corynebacterium is a Gram-positive bacterium and the most important GRAS microorganism for the production of certain amino-acids on an industrial scale, with a high potential for producing hyaluronic acid. Corynebacte
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