Large scale purification of an almond oleosin using an organic solvent procedure

Oleosins are small proteins which are specific to oil-bodies, the plant organelles specialized in oil storage. These proteins are thought to stabilize the structure and to prevent the oil droplets from coalescing. Since oleosins are insoluble in water, they are usually purified either as aggregates...

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Bibliographic Details
Published in:Plant physiology and biochemistry 2001-07, Vol.39 (7), p.623-630
Main Authors: Beisson, Frédéric, Ferté, Natalie, Voultoury, Robert, Arondel, Vincent
Format: Article
Language:English
Subjects:
ALMENDRA
almond
ALMONDS
AMANDE
Analytical, structural and metabolic biochemistry
Biological and medical sciences
emulsifying agent
emulsion
EXTRACCION POR DISOLVENTES
EXTRACTION PAR SOLVANT
Fundamental and applied biological sciences. Psychology
Life Sciences
METHODE
METHODS
METODOS
Miscellaneous
OIL CROPS
oil-body
oleosin
oleosome
PLANTAS OLEAGINOSAS
PLANTE OLEAGINEUSE
PROTEINAS
PROTEINE
PROTEINS
PRUNUS DULCIS
PURIFICACION
PURIFICATION
SOLVENT EXTRACTION
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Summary:Oleosins are small proteins which are specific to oil-bodies, the plant organelles specialized in oil storage. These proteins are thought to stabilize the structure and to prevent the oil droplets from coalescing. Since oleosins are insoluble in water, they are usually purified either as aggregates in water or as polypeptides solubilized with SDS. These conditions raise major problems for performing biophysical studies, however. We have taken advantage of the solubility of almond oleosin in chloroform to purify it by differential extraction using solvent mixtures. Detergent-free oleosin was purified to apparent homogeneity with a purification yield of 20 % using almond meal as starting material. An oil-body suspension was reconstituted by sonicating the solvent-purified oleosin with phospholipids and triacylglycerols. Protease protection experiments suggest that oleosins are inserted into the reconstituted oil-body as they are in native organelles. This simple purification procedure allows to obtain a detergent-free solution of oleosin easy to handle which can be used for oil-body reconstitution and biophysical studies by the Wilhemly plate method.
ISSN:0981-9428
1873-2690
0981-9428
DOI:10.1016/S0981-9428(01)01275-X