Relationship between acrylamide concentration and enzymatic activity in an improved single fibrin zymogram gel system.
Based on the zymography analysis, Bacillus sp. DJ-4 (screened from Doen-Jang, a Korean traditional fermented food) secretes seven extracellular fibrinolytic enzymes (EFEs; 68, 64, 55, 45, 33, 27, and 13 kDa) in culture broth. These seven EFEs were analyzed by newly applied SDSfibrin zymography combined with gradient polyacrylamide (SDS-FZGP). This improved gel system was used with a 5-20% acrylamide gradient in a fibrin zymogram gel for the separation of proteins with molecular masses from below 10 kDa to over 100 kDa on one gel plate. Using this system, high molecular weight bands (HMWBs) were clearly and sharply resolved. We also examined the relationship between an acrylamide concentration and the enzymatic activity of EFE using densitometric analysis.
Choi
Nack-Shick
NS
Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology, Yusong, Taejon 305-600, Korea.
Kim
Byoung-Young
BY
Lee
Jin-Young
JY
Yoon
Kab-Seog
KS
Han
Kyoung-Yoen
KY
Kim
Seung-Ho
SH
D020106:Acrylamide; D000818:Animals; D001407:Bacillus / Q000201:enzymology; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D005337:Fibrin; D005342:Fibrinolysis; D005516:Food Microbiology; D066298:In Vitro Techniques; D012967:Sodium Dodecyl Sulfate
D020106:Acrylamide; D012967:Sodium Dodecyl Sulfate; D005337:Fibrin; D010450:Endopeptidases