Food Science and Biotechnology

Korean Society of Food Science and Technology (한국식품과학회)
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Heterologous Gene Expression of aprE2 Encoding a 29 kDa Fibrinolytic Enzyme from Bacillus subtilis in Bacillus licheniformis ATCC 10716

  • Kwon, Gun-Hee (Division of Applied Life Science (BK21 Program), Gyeongsang National University) ;
  • Jeong, Woo-Ju (Division of Applied Life Science (BK21 Program), Gyeongsang National University) ;
  • Lee, Ae-Ran (Division of Applied Life Science (BK21 Program), Gyeongsang National University) ;
  • Park, Jae-Yong (Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Cha, Jae-Ho (Department of Microbiology, Pusan National University) ;
  • Song, Young-Sun (School of Food and Life Science, Inje University) ;
  • Kim, Jeong-Hwan (Division of Applied Life Science (BK21 Program), Gyeongsang National University)
  • Published : 2008.12.31
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Abstract

The aprE2 gene from Bacillus subtilis CH3-5 was expressed in Bacillus licheniformis ATCC 10716 using a Bacillus-Escherichai coli shuttle vector, pHY300PLK. The fibrinolytic activity of transformant (TF) increased significantly compared to B. licheniformis 10716 control cell. During the 100 hr incubation in Luria-Bertaini broth at $37^{\circ}C$, fibrinolytic activity of B. licheniformis TF increased rapidly at the late growth stage, after 52 hr of incubation, which was confirmed by zymography using a fibrin gel. pHY3-5 was stably maintained in B. licheniformis without tetracycline (Tc) in the media, 60.9% of cells still maintained pHY3-5 after 100 hr of cultivation.

Keywords

References

  1. Ageitos JM, Vallejo JA, Sestelo ABF, Poza M, Villa TG. Purification and characterization of a milk-clotting protease from Bacillus licheniformis strain USC13. J. Appl. Microbiol. 103: 2205-2213 (2007) https://doi.org/10.1111/j.1365-2672.2007.03460.x
  2. Creusot N, Gruppen H. Hydrolysis of whey protein isolate with Bacillus licheniformis protease: Fractionation and identification of aggregating peptides. J. Agr. Food Chem. 55: 9241-9250 (2007) https://doi.org/10.1021/jf071584s
  3. Schallmey M, Singh A, Ward OP. Developments in the use of Bacillus species for industrial production. Can. J. Microbiol. 50: 1-17 (2004) https://doi.org/10.1139/w03-076
  4. Peng Y, Yang X, Zhang Y. Microbial fibrinolytic enzymes: An overview of source, production, properties, and thrombolytic activity in vivo. Appl. Microbiol. Biot. 69: 126-132 (2005) https://doi.org/10.1007/s00253-005-0159-7
  5. Hwang K-J, Choi K-H, Kim M-J, Park CS, Cha J. Purification and characterization of a new fibrinolytic enzyme of Bacillus licheniformis KJ-31, isolated from Korean traditional jeotgal. J. Microbiol. Biotechn. 17: 1469-1476 (2007)
  6. Sumi H, Hamada H, Tsushima H, Mihara H, Muraki H. A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese natto; a typical and popular soybean food in the Japanese diet. Experientia 43: 1110-1111 (1987) https://doi.org/10.1007/BF01956052
  7. Kim WK, Choi K-H, Kim Y-T, Park H-H, Choi J-Y, Lee Y-S, Oh H-I, Kwon I-B, Lee S-Y. Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. strain CK 11-4 screened from cheonggukjang. Appl. Environ. Microb. 62: 2482-2488 (1996)
  8. Jeong S-J, Kwon G-H, Chun J, Kim JS, Park C-S, Kwon DY, Kim JH. Cloning of fibrinolytic enzyme gene from Bacillus subtilis isolated from cheonggukjang and its expression in protease-deficient Bacillus subtilis strains. J. Microbiol. Biotechn. 17: 1018-1023 (2007)
  9. Noh K-Y, Kim D-H, Choi N-S, Kim S-H. Isolation of fibrinolytic enzyme producing strains from kimchi. Korean J. Food Sci. Technol. 31: 219-223 (1999)