Synthesis activity-based zymography for detection of lipases and esterases.
A new zymography method for lipases and esterases was developed on the basis of the esterification reaction between fatty acids and alcohols. The enzymes were separated by SDS-PAGE and native PAGE. The gel was washed and then incubated in an aqueous solution containing fatty acids (oleic acid 18:1 or caprylic acid 8:0) and dodecanol. Synthesis was visualized by in situ precipitation of water-insoluble and non-diffusible fatty acid esters, such as dodecyl oleate and dodecyl octanoate. The synthesis activity-based zymography was confirmed with different enzyme samples, including commercial lipase preparations, purified recombinant lipase and cutinase, and crude culture supernatants of lipolytic enzyme-producing soil bacteria.
Chemical Biotechnology Research Center, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea.
Min-A
MA
Kwon
Hyun Suk
HS
Kim
Dae-Hyun
DH
Hahm
Jae Kwang
JK
Song
D001419:Bacteria / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D004950:Esterases / Q000032:analysis* / Q000302:isolation & purification; D008049:Lipase / Q000032:analysis* / Q000302:isolation & purification; D012988:Soil Microbiology
D004950:Esterases; D008049:Lipase