Reverse micellar extraction of lactoferrin from its synthetic solution using CTAB/n-heptanol system

Abstract

The partitioning of Lactoferrin (LF) into the reverse micellar phase formed by a cationic surfactant, cetyltrimethylammonium bromide (CTAB) in -heptanol from the synthetic solution of LF was studied. The solubilization behaviour of LF into the reverse micellar phase and back extraction using a fresh stripping phase were improved by studying the effect of processing parameters, including surfactant concentration, solution pH, electrolyte salt concentration and addition of alcohol as co-solvent. Forward extraction of 100% was achieved at CTAB concentration of 50 mM in-heptanol solvent, pH of 10 and 1 M NaCl. The electrostatic force and hydrophobic interaction have major influence on LF extraction during forward and back extraction respectively. The size of the reverse micelles and their corresponding water content were measured at different operating conditions to assess their role on the LF extraction. The present reverse micellar system has potential to solubilise almost all the LF into the reverse micelles during forward extraction and could able to allow back extraction from the reverse micellar phase with addition of small amount of co-solvent.

Effect of Key Parameters on Jacalin Extraction from Aqueous Phase into Anionic Reverse Micelles

“…However, further increase of the Vorg/Vaq ratio resulted in a decrease in protein transfer. This result is in line with the observations reported by other researchers which found the efficiency of extraction was decrease with larger volume ratio of organic to aqueous phase beyond 1:1, caused by the change of AOT concentration in the total mixture [35].…”

Selective extraction of lactoferrin from acidic whey using CTAB/n-heptanol reverse micellar system

“…The variation in the concentration of metals and other impurities in the whey may significantly influence the selective extraction of LF. The selective solubilization of LF from the synthetic whey protein solution, which was prepared by mixing the pure whey proteins, namely, a-LA, b-LG, LF, BSA, and LPO, into the organic phase was carried out at the reported optimized condition (50 mM CTAB in n-heptanol at pH 10 with the addition of 1 M NaCl) (Pawar et al 2017). Further, the experiments were also conducted for the acidic bovine whey, which was prepared in the laboratory.…”

“…Back extraction was carried out by introducing an equal volume of fresh aqueous stripping phase containing 1.3 M KCl as an additive and 7% V/V n-butanol as co-solvents to the organic phase obtained from forward extraction. The phases were mixed at 800 rpm for 1 h using magnetic stirrer and centrifuged at 5000 g for 30 min (Pawar et al 2017). After the phase separation, the phases are subjected to LF analysis.…”

“…LF occurs in other biological fluids like saliva, seminal plasma and tears in very less concentration compared to milk and whey (Pawar et al 2017). Even though the raw material is easily available and cheap, the cost of the pure LF in the open market is high due to the difficulties associated with the purification process, which has multiple process steps.…”

“…Recently, the authors studied the extraction of LF from its synthetic solution using cationic reverse micelles formed by 50 mM CTAB in n-heptanol at pH 10 with the addition of 1 M NaCl and obtained 100% entrapment of protein in micellar phase. The 98% of LF was back-extracted into the aqueous stripping phase at a pH of 6 in the presence of additives (7% n-propanol or n-butanol) and electrolyte (1.3 M KCl) (Pawar et al 2017). The suitability of the reported system is studied by extending the obtained process conditions for the selective extraction of LF from model whey proteins solution as well as acidic bovine whey.…”