Design of aqueous two-phase systems for purification of hyaluronic acid produced by metabolically engineeredLactococcus lactis
| Autor: | Vivek Rajendran, Kirubhakaran Puvendran, Bharath Raja Guru, Guhan Jayaraman |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0106 biological sciences
0301 basic medicine Aqueous solution Chromatography biology Membrane fouling Lactococcus lactis Filtration and Separation Polyethylene glycol biology.organism_classification 01 natural sciences Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Diafiltration 030104 developmental biology chemistry Biochemistry Potassium phosphate 010608 biotechnology Hyaluronic acid Fermentation |
| Zdroj: | Journal of Separation Science. 39:655-662 |
| ISSN: | 1615-9306 |
| DOI: | 10.1002/jssc.201500907 |
| Popis: | 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 of membrane 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. |
| Databáze: | OpenAIRE |
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