Multiple-layer substrate zymography for detection of several enzymes in a single sodium dodecyl sulfate gel.
We have developed a system to detect three hydrolytic enzymes (cellulase, lipase, and protease) using a single sodium dodecyl sulfate (SDS) gel and an electrotransfer system. After electrophoresis, proteins in the gel were transferred to three sandwiched substrate gels containing glycerol tributyrate, azo-carboxymethyl cellulose (Azo-CMC), and fibrin for detection of cellulase, lipase, and protease, respectively. We show that three cellulases (from a Paenibacillus sp. and two Bacillus sp. strains), one lipase (from a Staphylococcus sp.), and two proteases (from two Bacillus sp. strains) can be detected simultaneously with our zymogram system.
Choi
Nack-Shick
NS
Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology, Sinjeong-dong, Jeonbuk, Republic of Korea.
Kim
Bo-Hye
BH
Park
Chang-Su
CS
Han
Yun Jon
YJ
Lee
Hye Won
HW
Choi
Jong Hyun
JH
Lee
Seung-Goo
SG
Song
Jae Jun
JJ
D001426:Bacterial Proteins / Q000302:isolation & purification; D002480:Cellulase; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004798:Enzymes / Q000302:isolation & purification*; D008049:Lipase; D008722:Methods; D010447:Peptide Hydrolases; D012107:Research Design
D001426:Bacterial Proteins; D004798:Enzymes; D008049:Lipase; D002480:Cellulase; D010447:Peptide Hydrolases