Liquid-liquid equilibrium in ATPS and their application in purification of recombinant collagen-like protein from Rhodopseudomonas palustris expressed in E.Coli system

• Main Authors: Awang, Norsyahidatul Azwa, Amid, Azura, Mohd Arshad, Zatul Iffah
• Format: Conference or Workshop Item
• Language: English
• Published: International Institute for Halal Research and Training (INHART) 2021
• Subjects: TP248.13 Biotechnology
• Online Access: http://irep.iium.edu.my/93335/1/93335_Liquid-liquid%20equilibrium%20in%20ATPS.pdf; http://irep.iium.edu.my/93335/; https://conference.iium.edu.my/vihasc/

The aqueous two-phase system is a purification technique for biomolecules that has been receiving considerable attention among researchers. It is a complex phenomenon that depends on various factors, such as molecular weight of the polymer, concentration of polymer and salt, temperature and pH. The present work aimed to develop binodal curves made up of different molecular weights of polyethylene glycol (PEG) (1500, 2000, 4000, 6000 and 8000 g/mol), potassium phosphate buffer and water as well as to evaluate their potential for purification of recombinant collagen-like protein (rCLP) from Rhosopseudomonas palustris. The result showed that a high molecular weight of PEG required a low concentration of potassium phosphate to form a two-phase system and, as the PEG molecular weight increased, the curve was distorted towards the origin. Furthermore, non-linear equations were satisfactorily fitted to the experimental data of binodal curves and the corresponding standard deviations for the systems were obtained. The ATPS with PEG2000/potassium phosphate buffer at pH 7.0 was found to be suitable for the recovery of rCLP from the cell lysate and led to preferential migration of molecules to the PEG-rich top phase.