title: Enhancement of sequential zymography technique for the detection of thermophilic lipases and proteases.
abstract: Analysis of lipases and proteases present in cell-free fractions of thermophilic Bacillus sp. cultures were performed in an enhanced sequential zymography method. After the PAGE run, the gel was electrotransferred to another polyacrylamide gel containing a mixture of glycerol tributyrate, olive oil and gelatin. After transference, this substrate-mix gel was incubated for lipase detection, until bands appeared, and later stained with CBB for protease detection. Assets are, besides detecting two enzymes on a single gel, time and material saving.
authors.affiliation: Center for Environmental, Biology and Chemistry Research, Facultad de Ciencias y Tecnología, Universidad de Carabobo, Apartado Postal 3433, Av. Salvador Allende, Valencia, 2001, Carabobo, Venezuela, jwilkesm@uc.edu.ve.
authors.forename: Jeff
authors.initials: J
authors.lastname: Wilkesman
authors.forename: Zully
authors.initials: Z
authors.lastname: Hernández
authors.forename: Marleny
authors.initials: M
authors.lastname: Fernández
authors.forename: Lellys M
authors.initials: LM
authors.lastname: Contreras
authors.forename: Liliana
authors.initials: L
authors.lastname: Kurz
mesh_terms: D001407:Bacillus / Q000737:chemistry / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry*; D057075:Enzyme Assays; D004795:Enzyme Stability; D006358:Hot Temperature; D008049:Lipase / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D008049:Lipase; D010447:Peptide Hydrolases