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Protein extraction using reverse micelles

Protein extraction using reverse micelles

Pham, Thi Minh Hai (2015) Protein extraction using reverse micelles. PhD thesis, University of Greenwich.

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Abstract

Reverse micelles are self-organized aggregates formed by a surfactant in a non-polar solvent or oil. The presence of a water pool in the polar core of reverse micelles is of considerable advantage in protein extraction. A lot of researches have been done with ionic reverse micelles applied in protein extraction. However, this ability of non-ionic reverse micelles has not been fully understood and therefore requires more research.
In this project, different surfactants (anionic AOT, cationic CTAB, non-ionic triblock L61 copolymer) were investigated for their ability to form RM and for their application in protein extraction. It was found that lysozyme could be extracted using an AOT RMS, but not with a CTAB RMS. For the first time, an aromatic solvent, p-xylene, was used for the extraction of lysozyme and it was found that the AOT in p-xylene RM system resulted in the higher lysozyme activity (73.81 %) compared to an AOT/isooctane RM system (43.2 %). The effect of different salts (KCl, KF, KBr) on the FE and BE of BSA was investigated using the CTAB in mixture of 1-bromooctane, 1-hexanol and petroleum ether. The results indicated that KCl gave the highest extraction efficiency of 64 % as compared to around 40 % with both cases of KF and KBr. The secondary structure of extracted BSA was maintained with KCl only. L61 pluronics polymers was investigated for its reverse micelles forming ability and it was established that small reverse micelles with a maximum W0 of 4 was formed. Because of the small size of L61 reverse micelles, lysozyme could not be extracted but was precipitated out when combined with the co-surfactant AOT. The activity of the recovered lysozyme from the precipitate was maintained (66% as compared with native lysozyme). Moreover, if L61 was used as a co-surfactant with AOT reverse micelles, extraction efficiency was improved (88 %) and the activity of the extracted ly-sozyme was increased (56 %) as compare to extraction with an AOT system alone (46 %). These studies thus gives useful insights in the role of individual and mixed surfactant systems in the extraction and precipitation of proteins.

Item Type: Thesis (PhD)
Uncontrolled Keywords: Reverse micellar system (RMS); Lysozyme extraction;
Subjects: Q Science > QD Chemistry
Faculty / Department / Research Group: Faculty of Engineering & Science
Faculty of Engineering & Science > Department of Pharmaceutical, Chemical & Environmental Sciences
Last Modified: 22 Nov 2017 12:52
Selected for GREAT 2016: None
Selected for GREAT 2017: None
Selected for GREAT 2018: None
URI: http://gala.gre.ac.uk/id/eprint/18122

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