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.