Cloning of fibrinolytic enzyme gene from Bacillus subtilis isolated from Cheonggukjang and its expression in protease-deficient Bacillus subtilis strains.
Bacillus subtilis CH3-5 was isolated from cheonggukjang prepared according to traditional methods. CH3-5 secreted at least four different fibrinolytic proteases (63, 47, 29, and 20 kDa) into the culture medium. A fibrinolytic enzyme gene, aprE2, encoding a 29 kDa enzyme was cloned from the genomic DNA of CH3-5, and the DNA sequence determined. aprE2 was overexpressed in heterologous B. subtilis strains deficient in extracellular proteases using a E. coli-Bacillus shuttle vector. A 29 kDa AprE2 band was observed and AprE2 seemed to exhibit higher activities towards fibrin rather than casein.
Division of Applied Life Science, Graduate School, Gyeongsang National University, Jinju 660-701, Korea.
Seon-Ju
SJ
Jeong
Gun-Hee
GH
Kwon
Jiyeon
J
Chun
Jong Sang
JS
Kim
Cheon-Seok
CS
Park
Dae Young
DY
Kwon
Jeong Hwan
JH
Kim
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics*; D001483:Base Sequence; D002364:Caseins / Q000378:metabolism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D005285:Fermentation; D005337:Fibrin / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D013025:Glycine max / Q000382:microbiology*
D002364:Caseins; D011994:Recombinant Proteins; D005337:Fibrin; D010447:Peptide Hydrolases