title: Properties of a Fibrinolytic Enzyme Secreted byBacillus amyloliquefaciensRSB34, Isolated from Doenjang.
abstract: Nine bacilli with fibrinolytic activities were isolated from doenjang, a traditional Korean fermented soy food. Among them, RSB34 showed the strongest activity and was identified asBacillus amyloliquefaciensby 16S rRNA andrecAgene sequencing. During growth on LB up to 96 h, RSB34 showed the highest fibrinolytic activity (83.23 mU/μl) at 48 h. Three bands of 23, 27, and 42 kDa in size were observed when the culture supernatant was analyzed by SDS-PAGE and 27 and 42 kDa bands by fibrin zymography. The gene encoding the 27 kDa fibrinolytic enzyme AprE34 was cloned by PCR. BLAST analyses confirmed that the gene was a homolog to genes encoding AprE-type proteases.aprE34was overexpressed inEscherichia coliBL21(DE3) using pET26b(+). Recombinant AprE34 was purified and examined for its properties. TheKmandVmaxvalues of recombinant AprE34 were 0.131 ± 0.026 mM and 16.551 ± 0.316 μM/l/min, respectively, when measured using an artificial substrate,N-succinyl-ala-ala-pro-phe-p-nitroanilide.aprE34was overexpressed inB. subtilisWB600 using pHY300PLK.B. subtilistransformants harboring pHYRSB34 (pHY300PLK withaprE34) showed higher fibrinolytic activity thanB. amyloliquefaciensRSB34.
authors: Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University, Jinju 52828, Republic of Korea.
authors: Zhuang
authors: Z
authors: Yao
authors: Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University, Jinju 52828, Republic of Korea.
authors: Xiaoming
authors: X
authors: Liu
authors: Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University, Jinju 52828, Republic of Korea.
authors: Jae Min
authors: JM
authors: Shim
authors: Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University, Jinju 52828, Republic of Korea.
authors: Kang Wook
authors: KW
authors: Lee
authors: Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University, Jinju 52828, Republic of Korea.|Institute of Agriculture and Life Sciences, Gyeongsang National University, Jinju 52828, Republic of Korea.
authors: Hyun-Jin
authors: HJ
authors: Kim
authors: Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University, Jinju 52828, Republic of Korea.|Institute of Agriculture and Life Sciences, Gyeongsang National University, Jinju 52828, Republic of Korea.
authors: Jeong Hwan
authors: JH
authors: Kim
keywords: Bacillus amyloliquefancies; doenjang; fibrinolytic enzymes; gene cloning