Design of aqueous two-phase systems for purification of hyaluronic acid produced by metabolically engineered Lactococcus lactis

Rajendran, Vivek and Puvendran, Kirubhakaran and Guru, Bharath Raja and Jayaraman, Guhan (2017) Design of aqueous two-phase systems for purification of hyaluronic acid produced by metabolically engineered Lactococcus lactis. Journal of Separation Science, 39 (4). pp. 655-662. ISSN 1615-9306

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Abstract

Hyaluronic acid has a wide range of biomedical applications and its commercial value is highly dependent on its purity and molecular weight. This study highlights the utility of aqueous two-phase separation as a primary recovery step for hyaluronic acid and for removal of major protein impurities from fermentation broths.Metabolically engineered cultures of a lactate dehydrogenase mutant strain of Lactococcus lactis (L. lactis NZ9020) were used to produce high-molecular-weight hyaluronic acid. The cell-free fermentation broth was partially purified using a polyethylene glycol/potassium phosphate system, resulting in nearly 100% recovery of hyaluronic acid in the salt-rich bottom phase in all the aqueous two-phase separation experiments. These experiments were optimized for maximum removal of protein impurities in the polyethylene glycol rich top phase. The removal of protein impurities resulted in substantial reduction ofmembrane fouling in the subsequent diafiltration process, carried out with a 300 kDa polyether sulfone membrane. This step resulted in considerable purification of hyaluronic acid, without any loss in recovery and molecular weight. Diafiltration was followed by an adsorption step to remove minor impurities and achieve nearly 100% purity. The final hyaluronic acid product was characterized by Fourier-transform IR and NMR spectroscopy, confirming its purity

Item Type: Article
Uncontrolled Keywords: Aqueous two-phase Systems / Diafiltration / Hyaluronic acid / Lactococcus lactis
Subjects: Engineering > MIT Manipal > Biotechnology
Depositing User: MIT Library
Date Deposited: 13 Jul 2017 09:01
Last Modified: 13 Jul 2017 09:01
URI: http://eprints.manipal.edu/id/eprint/149316

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