Capture of sunflower seedlings lipase using polyclonal antibodies.
True lipolytic activity is observed in different subcellular fractions of germinating sunflower seedlings (Helianthus annuus L.) in delipidated oleosomes and microsomes. Triacylglycerol lipase (EC. 3.1.1.3) catalyses the first catabolic step of lipolysis. To our knowledge, this plant lipase has not yet been identified. Our aim was to develop a method to collect the lipase for further studies. An immunological method was used to capture sunflower seedling lipase from oleosomes and microsomes. This method uses an immunoaffinity column prepared with polyclonal antibodies (anti-P-61) directed against oleosomal activity. Our results verify that we have successfully adapted a purification procedure of plant lipase using anti-P-61. Since the eluted lipolytic activity is distributed among diverse proteic peaks, we changed the elution procedure: the introduction of CHAPS, a zwitterionic detergent, allowed us to recover all the lipolytic activity in a single proteic peak. This may help us to characterise the studied lipase.
Laboratoire de Biochimie des protéines, Département des Sciences Biologiques, Faculté des Sciences de Tunis, Campus Universitaire, Tunis, Tunisia. sellemab@gmail.com
Sellema
S
Bahri
Raja
R
Marrakchi
Amel
A
Benammar-Elgaaied
Jeannette
J
Ben Hamida
D004797:Enzyme-Linked Immunosorbent Assay; D006368:Helianthus / Q000201:enzymology*; D007158:Immunologic Techniques; D008049:Lipase / Q000276:immunology* / Q000302:isolation & purification* / Q000378:metabolism; D008861:Microsomes / Q000276:immunology; D015388:Organelles / Q000201:enzymology; D036226:Seedlings; D013347:Subcellular Fractions
D008049:Lipase