title: Purification and characterization of a major fibrinolytic enzyme from Bacillus amyloliquefaciens MJ5-41 isolated from Meju.
abstract: Meju is a traditional Korean fermented soy product used as a key element for soy sauce and doenjang. Bacilli with antimicrobial activity were isolated from meju prepared by traditional methods at Sunchang county, Jeollabukdo, Korea. Six isolates were identified as Bacillus amyloliquefaciens by recA gene sequencing and RAPD-PCR. One isolate, B. amyloliquefaciens MJ5-41, showed the strongest fibrinolytic activity. A 27 kDa active fibrinolytic enzyme, AprE5-41, was purified from the culture supernatant of MJ5-41 grown on LB by chromatographic methods. The optimum pH and temperature for purified AprE5-41 were 7.0 and 45°C, respectively. AprE5-41 quickly degraded Aα and Bβ chains but not the gamma-chain of fibrinogen. AprE5-41 exhibited the highest specificity for N-succinyl-Ala-Ala-Pro-Phe pnitroanilide, a known substrate for α-chymotrypsin, cathepsin G, and subtilisin BPN'. The structural gene, aprE5-41, was cloned by PCR and successfully expressed in B. subtilis.
authors: Division of Applied Life Science (BK21 program), Graduate School, Gyeongsang National University, Jinju 660-701, Korea.
authors: Hyeon-Deok
authors: HD
authors: Jo
authors: Hwang A
authors: HA
authors: Lee
authors: Seon-Ju
authors: SJ
authors: Jeong
authors: Jeong Hwan
authors: JH
authors: Kim
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004795:Enzyme Stability; D005337:Fibrin / Q000378:metabolism; D005341:Fibrinolysin / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D005516:Food Microbiology; D006863:Hydrogen-Ion Concentration; D007723:Korea; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019105:Random Amplified Polymorphic DNA Technique; D011938:Rec A Recombinases / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D004269:DNA, Bacterial; D005337:Fibrin; D011938:Rec A Recombinases; D005341:Fibrinolysin