Single-step purification of different lipases from Staphylococcus warneri.
Three different lipases from the extract crude of Staphylococcus warneri have been purified by specific lipase-lipase interactions using different lipases (TLL, RML, PFL, BTL2) covalently attached to a solid support as adsorption matrix. BTL2 immobilized on glyoxyl-DTT adsorbed selectivity only a 30 kDa lipase from the crude, which was desorbed by adding 0.1% triton X-100. Using glyoxyl-PFL as matrix, two new lipases (28 and 40 kDa) were adsorbed, and completely pure 40 kDa lipase was obtained after desorption using 0.01% triton, whereas 28 kDa lipase was desorbed after the incubation of the lipase matrix with 3% detergent. When using other matrixes as glyoxyl-TLL or glyoxyl-RML, different lipases were adsorbed. This methodology could be a very efficient and useful method to purify several lipases from crude extracts from different sources.
Volpato
Giandra
G
Departamento de Biocatalisis, Instituto de Catalisis (CSIC), Madrid, Spain.
Filice
Marco
M
Ayub
Marco A Z
MA
Guisan
Jose M
JM
Palomo
Jose M
JM
D000327:Adsorption; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D006038:Glyoxylates / Q000737:chemistry; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008958:Models, Molecular; D011485:Protein Binding; D012685:Sepharose / Q000737:chemistry; D013210:Staphylococcus / Q000201:enzymology*
D004800:Enzymes, Immobilized; D006038:Glyoxylates; C070399:glyoxyl agarose; D012685:Sepharose; D008049:Lipase