Improvement of Fibrinolytic Activity of Bacillus subtilis 168 by Integration of a Fibrinolytic Gene into the Chromosome.

  title={Improvement of Fibrinolytic Activity of Bacillus subtilis 168 by Integration of a Fibrinolytic Gene into the Chromosome.},
  author={Seon-Ju Jeong and Ji Yeong Park and Jae Yong Lee and Kang-Wook Lee and Kye Man Cho and Gyoung Min Kim and Jung-Hye Shin and Jong-Sang Kim and Jeong Hwan Kim},
  journal={Journal of microbiology and biotechnology},
  volume={25 11},
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… 
4 Citations

Figures and Tables from this paper

Increase of a Fibrinolytic Enzyme Production through Promoter Replacement of aprE3-5 from Bacillus subtilis CH3-5.

Results confirmed that significant overproduction of a fibrinolytic enzyme can be achieved by suitable promoter modification, and this approach may have applications in the industrial production of AprE3-5 and related fibrin zymography enzymes.

Cloning, Secretory Expression, Partial Characterization, and Structural Modeling of an Alkaline Protease from Bacillus subtilis D-2

The data suggested that the secretory expression system with pET-32a in E. coli was successfully constructed and enzymatic specificity analysis of the alkaline protease indicated that it was suitable for various processing industries.

Production, purification, and functional properties of microbial fibrinolytic enzymes produced by microorganism obtained from soy-based fermented foods: developments and challenges.

According to the World Health Organization, cardiovascular disease (CVD) has become a major cause of chronic illness around the globe. It has been reported that soy-based fermented food (SFF) is very

Current Perspectives on the Physiological Activities of Fermented Soybean-Derived Cheonggukjang

The physiological activities of CGJ were examined and its potential as a functional food was confirmed, as it can be made without salt, unlike other pastes.



Characterization of AprE176, a fibrinolytic enzyme from Bacillus subtilis HK176.

Molecular modeling analysis suggested that the 176(th) residue of M179, threonine, was located near the cation-binding site compared with the wild type, which probably caused tight binding of M 179 with Ca(2+), which increased the thermostability of M178.

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 and aprE2 seemed to exhibit higher activities towards fibrin rather than casein, and the DNA sequence determined.

Characterization of a 27 kDa fibrinolytic enzyme from Bacillus amyloliquefaciens CH51 isolated from cheonggukjang.

Among four different growth media, tryptic soy broth was the best in terms of supporting cell growth and fibrinolytic activity of this strain of Bacillus amyloliquefaciens CH51, and it was accordingly named AprE51.

Microbial fibrinolytic enzymes: an overview of source, production, properties, and thrombolytic activity in vivo

Accumulation of fibrin in the blood vessels usually results in thrombosis, leading to myocardial infarction and other cardiovascular diseases. For thrombolytic therapy, microbial fibrinolytic enzymes

A new method for multiple gene inactivations in Bacillus subtilis 168, producing a strain free of selectable markers.

A novel method for repeated gene inactivation in Bacillus subtilis 168 is described, which inactivated nprE and aprE, two protease genes secreted by B. subtil is 168 free of selectable markers.

Developments in the use of Bacillus species for industrial production.

Developments from the understanding of the complex protein translocation machinery of Gram-positive bacteria should allow the resolution of current secretion challenges and make Bacillus species preeminent hosts for heterologous protein production.

Site-directed mutagenesis and over expression of aroG gene of Escherichia coli K-12.

Bacillus fermentation of soybeans

The fermentations containing E. faecium and Bacillus exhibited a greater initial pH decline and a slightly retarded subseqent increase in pH compared with fermentations with Bacillus only, and the presence of E. Faecium had no detectable effects on growth of the Bacillus, proteolytic activity, ammonia production or the final pH of the fermentations.

Development of a New Integration Site within theBacillus subtilis Chromosome and Construction of Compatible Expression Cassettes

It was shown that the integration of a regulatory gene at the lacA locus was able to control the expression of a transcriptional fusion at the amyE locus, demonstrating that both integration sites can be used simultaneously to obtain regulated expression of desired genes.

A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese Natto; a typical and popular soybean food in the Japanese diet

A novel fibrinolytic enzyme, named nattokinase, was demonstrated in the vegetable cheese Natto, which is a typical soybean food eaten in Japan, and it digested fibrin but also the plasmin substrate H-D-Val-Leu-Lys-pNA, which was more sensitive to the enzyme than other substrates tried.