title: Improvement of Fibrinolytic Activity of Bacillus subtilis 168 by Integration of a Fibrinolytic Gene into the Chromosome.
abstract: Fibrinolytic enzyme genes (aprE2, aprE176, and aprE179) were introduced into the Bacillus subtilis 168 chromosome without any antibiotic resistance gene. An integration vector, pDG1662, was used to deliver the genes into the amyE site of B. subtilis 168. Integrants, SJ3-5nc, SJ176nc, and SJ179nc, were obtained after two successive homologous recombinations. The integration of each fibrinolytic gene into the middle of the amyE site was confirmed by phenotypes (Amy(-), Spec(S)) and colony PCR results for these strains. The fibrinolytic activities of the integrants were higher than that of B. subtilis 168 by at least 3.2-fold when grown in LB broth. Cheonggukjang was prepared by inoculating each of B. subtilis 168, SJ3-5nc, SJ176nc, and SJ179nc, and the fibrinolytic activity of cheonggukjang was 4.6 ± 0.7, 10.8 ± 0.9, 7.0 ± 0.6, and 8.0 ± 0.2 (U/g of cheonggukjang), respectively at 72 h. These results showed that construction of B. subtilis strains with enhanced fibrinolytic activities is possible by integration of a strong fibrinolytic gene via a marker-free manner.
authors.affiliation: Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Seon-Ju
authors.initials: SJ
authors.lastname: Jeong
authors.affiliation: Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Ji Yeong
authors.initials: JY
authors.lastname: Park
authors.affiliation: Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Jae Yong
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Kang Wook
authors.initials: KW
authors.lastname: Lee
authors.affiliation: Department of Food Science, Gyeongnam National University of Science and Technology, Jinju 660-758, Republic of Korea.
authors.forename: Kye Man
authors.initials: KM
authors.lastname: Cho
authors.affiliation: Namhae Garlic Research Institute, Namhae 668-812, Republic of Korea.
authors.forename: Gyoung Min
authors.initials: GM
authors.lastname: Kim
authors.affiliation: Namhae Garlic Research Institute, Namhae 668-812, Republic of Korea.
authors.forename: Jung-Hye
authors.initials: JH
authors.lastname: Shin
authors.affiliation: School of Food Science and Biotechnology (BK21 Plus), Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Jong-Sang
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University, Jinju 660-701, Republic of Korea.|Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Jeong Hwan
authors.initials: JH
authors.lastname: Kim
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D002876:Chromosomes, Bacterial; D003470:Culture Media / Q000737:chemistry; D005341:Fibrinolysin / Q000235:genetics* / Q000378:metabolism*; D005342:Fibrinolysis; D005798:Genes, Bacterial; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins; D005341:Fibrinolysin
keywords: Bacillus subtilis; chromosome integration; fibrinolytic genes; integration vector