Purification of different extracellular lipases from Geotrichum candidum 4013 by using a simple strategy

0394851 - ÚOCHB 2014 GB eng A - Abstrakt
Brabcová, Jana - Zarevúcka, Marie - Palomo, J. M.
Purification of different extracellular lipases from Geotrichum candidum 4013 by using a simple strategy.
BIOTRANS 2013. Manchester: -, 2013. s. 105-106.
[BIOTRANS 2013. 21.07.2013-25.07.2013, Manchester]
Grant ostatní: AV ČR(CZ) M200551203
Institucionální podpora: RVO:61388963
Klíčová slova: lipase * purification * activity
Kód oboru RIV: CC - Organická chemie

Three different lipases from crude preparations of Geotrichum candidum 4013 have been purified in a simple fashion. A crude medium with extracellular lipases was offered to octyl agarose. The support specifically adsorbed two lipases. A completely pure new 45 kDa lipase was obtained after desorption using 0.4% Triton X-100, whereas 61 kDa lipase was desorbed after the incubation of the lipase matrix with 1% of this detergent. The addition of octadecyl-Sepabeads support to the non-adsorbed proteins on octyl-agarose permitted to selectively adsorb a third lipase, having a molecular weight of 59 kDa. Desorption of the enzyme using 1% Triton X-100 permitted to have also a pure sample of this enzyme as confirmed SDS-PAGE. These enzymes showed a lipolytic activity in zymography assay with methylumbelliferone butyrate as substrate. The highest yield of hydrolysis of racemic ester trans-2-(4-methoxybenzyl)-1-cyclohexanol was obtained by new lipase 45 kDa. The identity of the peptides was confirmed by using mass spectrometry and the primary sequence of N-terminal region of the novel lipase with molecular weight of 45 kDa was determined.
Trvalý link: http://hdl.handle.net/11104/0223112