Extractive purification of recombinant thermostable lipase from fermentation broth of Escherichia coli using an aqueous polyethylene glycol impregnated resin system
This study aimed at recovery of thermostable lipase from Escherichia coli BL21 using porous glass beads grafted with polyethylene glycol (PEG) in aqueous impregnated resins system (AIRS). The influencing parameters such as concentration and pH of extraction solution, concentration of NaCl, size of the beads, and pH of the desorption solution on the partition behaviour of lipase were evaluated. Smaller adsorbent (4 mm) had a 65.5% of recovery yield with approximately two-fold higher purification factor compared to that obtained with the larger adsorbent. Recombinant lipase was purified successfully using AIRS with a purification factor of 7.6 and yield of 78.4% under optimum conditions of 18% (w/w) PEG 4000, 10% (w/w) of potassium citrate at pH 9 with 3% (w/w) of NaCl. Optimum desorption was obtained with 4.0 mm of porous glass beads at pH 9.
KeywordsAqueous impregnated resins system Aqueous phase Purification Recovery Lipase
This work was funded by Universiti Sains Malaysia under USM Short Term Grant (PO4865, 304/PTENKIND/6315138).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
- Gedikli M, Ceylan Ş, Erzengin M, Odabaşı M (2014) A novel matrix for hydrophobic interaction chromatography and its application in lysozyme adsorption. Acta Biochim Pol 61(4):731–737Google Scholar
- Ketnawa S, Rungraeng N, Rawdkuen S (2017) Phase partitioning for enzyme separation: an overview and recent applications. Int Food Res J 24(1):1–24Google Scholar
- Ramyadevi D, Subathira A, Saravanan S (2012) Aqueous two-phase poly (ethylene glycol)–Maltodextrin system for protein partitioning from shrimp waste: influence of molecular weight and pH. Int J Environ Sci 2(4):2462–2469Google Scholar
- Shabram P, Vellekamp G, Ruan Q, Scandella C (2002) Purification of adenovirus. In: Curiel DT (ed) Adenoviral vectors for gene therapy, 2nd edn. Academic Press, San Diego, pp 197–230Google Scholar
- Zafarani-Moattar MT, Hamzehzadeh S (2011) Effect of pH on the phase separation in the ternary aqueous system containing the hydrophilic ionic liquid 1-butyl-3-methylimidazolium bromide and the kosmotropic salt potassium citrate at T = 298.15 K. Fluid Phase Equilib 304(1):110–120CrossRefGoogle Scholar