Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
권호 표지

Isolation, Identification, and Characterization of Bacillus strains from the Traditional Korean Soybean-fermented Food, Chungkookjang

  • Joo, Myeong-Hoon (Graduate School of Molecular & Biomedical Technology, Inje University) ;
  • Hur, Sung-Ho (Department of Clinical Laboratory Science, Dong-eui Institute of Technology) ;
  • Han, Yong-Soo (Department of Cosmetology, Gwangyang Health College) ;
  • Kim, Ji-Yeon (Graduate School of Molecular & Biomedical Technology, College of General Education, Inje University)
  • Published : 2007.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

A total of 45 bacterial strains were isolated from the traditional Korean soybean-fermented food, Chungkookjang. Among these strains, seven strains were selected and identified based on morphological, physiological, and biochemical characteristics, as well as phylogenetic analysis using 16S rDNA sequences. All strains were Gram-positive, aerobic, motile, oxidase-positive, rod-shaped, and endospore-forming bacteria, and produced extracellular enzymes such as amylase, cellulase, lipase, protease, and xylanase. The isolates were grown in the presence of 0-11% (w/v) NaCl. Growth was optimal at pH 6-9 and at temperatures of $30-45^{\circ}C$. According to VITEK automicrobic system tests and supplementary tests, the isolates were similar to several species of the genus Bacillus. The phylogenetic analysis of seven bacterial strains based on comparisons of 16S rDNA sequences, revealed that the strains were closely related to Bacillus species. The identification of strains that produced surfactin was also carried out, based on PCR screening of the sfp gene. Among the seven isolated strains, six yielded a surfactin-positive result with PCR.

Keywords

References

  1. Agrawal M, Pradeep S, Chandraraj K, and Gummadi SN (2005) Hydrolysis of starch by amylase from Bacillus sp. KCA102: a statistical approach. Process Biochem 40, 2499-2507 https://doi.org/10.1016/j.procbio.2004.10.006
  2. Ali Amoozegar M, Zahra Fatemi A, Reza Karbalaei-heidari H, and Reza Razavi M (2006) Production of an extracellular alkaline metalloprotease from a newly isolated, moderately halophile, Salinivibrio sp. strain AF-2004. Microbiol Res (Article) 1-9
  3. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, and Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25, 3389-3402 https://doi.org/10.1093/nar/25.17.3389
  4. Ashiuchi M, Kamei T, Baek DH, Shin SY, Sung MH, Soda K, Yagi T, and Misono H (2001) Isolation of Bacillus subtilis (chungkookjang), a poly-${\gamma}$-glutamate procedure with high genetic competence. Appl Microbiol Biotechnol 57, 764-769 https://doi.org/10.1007/s00253-001-0848-9
  5. Banat IM, Makkar RS, and Cameotra SS (2000) Potential commercial applications of microbial surfactants. Appl Microbiol Biotechnol 53, 495-508 https://doi.org/10.1007/s002530051648
  6. Benson DA, Boguski MS, Lipman DJ, Ostell J, Oullette BFF, Rapp BA, and Wheeler DL (1999) GenBank. Nucleic Acids Res 27, 12-17 https://doi.org/10.1093/nar/27.1.12
  7. Besson F and Michel G (1992) Biosynthesis of iturin and surfactin by Bacillus subtilis: Evidence for amino acid activating enzymes. Biotechnol Lett 14, 1013-1018 https://doi.org/10.1007/BF01021050
  8. Carrillo PG, Mardaraz C, Pitta-Alvarez SI, and Giulietti AM (1996) Isolation and selection of biosurfactant-producing bacteria. World J Microbiol Biotechnol 12, 82-84 https://doi.org/10.1007/BF00327807
  9. Cherry JR and Fidantsef AL (2003) Directed evolution of industrial enzymes: an update. Curr Opin Biotechnol 14, 438-443 https://doi.org/10.1016/S0958-1669(03)00099-5
  10. Claus D and Berkeley RCW (1986) Genus Bacillus. In Bergey's Manual of Systematic Bacteriology, pp. 1105- 1139, Williams & Wilkins, Baltimore
  11. Desai JD and Banat IM (1997) Microbial production of surfactants and their commercial potential. Microbiol Mol Biol Rev 61, 47-64
  12. Diaz P, Prim N, and Pastor FIJ (1999) Direct fluorescencebased lipase activity assay. Biotechniques 27, 697-699
  13. Feignier C, Besson F, and Michel G (1995) Studies on lipopeptide biosynthesis by Bacillus subtilis: isolation and characterization of iturin-, surfactin + mutants. FEMS Microbiol Lett 127, 11-15
  14. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39, 783-791 https://doi.org/10.2307/2408678
  15. Hsieh FC, Li MC, Lin TC, and Kao SS (2004) Rapid detection and characterization of surfactin-producing Bacillus subtilis and closely related species based on PCR. Curr Microbiol 49, 186-191
  16. Hwang MH, Lim JH, Yun HI, Rhee MH, Cho JY, Hsu WH, and Park SC (2005) Surfactin C inhibits the lipopolysaccharide- induced transcription of interleukin-1 and inducible nitric oxide synthase and nitric oxide production in murine RAW 264.7 cells. Biotechol Lett 27, 1605-1608 https://doi.org/10.1007/s10529-005-2515-1
  17. Kang MJ, Kim JI, and Kwon TW (2003) Effect of Chongkukjang on blood glucose and lipid profile in neonatal streptozotocin-induced diabetic rats. Food Sci Biotechnol 12, 544-547
  18. Karanth NGK, Deo PG, and Veenanadig NK (1999) Microbial production of biosurfactants and their importance. Curr Sci 77, 116-126
  19. Kim K, Jung SY, Lee DK, Jung JK, Park JK, Kim DK, and Lee CH (1998) Supression of inflammatory responses by surfactin, a selective inhibitor of platelet cytosolic phopholipase A2. Biochem Pharmacol 55, 975-985 https://doi.org/10.1016/S0006-2952(97)00613-8
  20. Kim SS, Lee JH, Ahn YS, Kim JH, and Kang DK (2003a) A fibrinolytic enzyme from Bacillus amyloliquefaciens D4-7 isolated from Chungkook-jang: It's characterization and influence of additive on thermostability. Korean J Microbiol Biotechnol 31, 271-276
  21. Kim Y, Cho JY, Kuk JH, Moon JH, Cho JI, Kim YC, and Park KH (2004) Identification and antimicrobial activity of phenylacetic acid produced by Bacillus licheniformis isolated from fermented soybean, Chungkook-Jang. Curr Microbiol 48, 312-317 https://doi.org/10.1007/s00284-003-4193-3
  22. Kim YS, Jung HJ, Park YS, and Yu TS (2003b) Characteristics of flavor and functionality of Bacillus subtilis K-20 Chungkukjang. Korean J Food Sci Technol 35, 475-478
  23. Ko JH, Yan JP, Zhu L, and Qi YP (2004) Identification of two novel fibrinolytic enzymes from Bacillus subtilis QK02. Comp Biochem Physiol C Toxicol Pharmacol 137, 65-74 https://doi.org/10.1016/j.cca.2003.11.008
  24. Kwon HY, Kim YS, Kwon GS, Kwon CS, and Sohn HY (2004) Isolation of immuno-stimulating strain Bacillus pumilus JB-1 from Chungkukjang and fermentational characteristics of JB-1. Korean J Microbiol Biotechnol 32, 291-296
  25. Lane DJ (1991) 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systematics, pp. 115-175, John Wiley & Sons, New York
  26. Lee CH, Yang EI, Song GS, Chai OH, and Kim YS (2005a) Chongkukjang mucilage stimulates immunohistochemical activities of gastrointestinal tract in rats. Food Sci Biotechnol 14, 813-817
  27. Lee EJ and Kim JK (2004) Characteristics of taste components of Chongkukjang fermented with Bacillus subtilis. Food Sci Biotechnol 13, 572-575
  28. Lee MY, Park SY, Jung KO, Park KY, and Kim SD (2005b) Quality and functional characteristics of Chungkukjang prepared with various Bacillus sp. isolated from traditional Chungkukjang. J Food Sci 70, M191-196
  29. Mulligan CN (2005) Environmental applications for biosurfactants. Environ Pollut 133, 183-198 https://doi.org/10.1016/j.envpol.2004.06.009
  30. Nakano MM, Corbell N, Besson J, and Zuber P (1992) Isolation and characterization of sfp: A gene that functions in the production of the lipopeptide biosurfactant, surfactin, in Bacillus subtilis. Mol Gen Genet 232, 313-321
  31. Nakano MM, Marahiel MA, and Zuber P (1988) Identification of a genetic locus required for biosynthesis of the lipopeptide antibiotic surfactin in Bacillus subtilis. J Bacteriol 170, 5662-5668 https://doi.org/10.1128/jb.170.12.5662-5668.1988
  32. Peypoux F, Bonmatin JM, and Wallacb J (1999) Recent trends in the biochemistry of surfactin. Appl Microbial Biotechnol 51, 553-563 https://doi.org/10.1007/s002530051432
  33. Rosenberg E and Ron EZ (1999) High- and low-molecularmass microbial surfactants. Appl Microbiol Biotechnol 52, 154-162 https://doi.org/10.1007/s002530051502
  34. Saitou N and Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic tree. Mol Biol Evol 4, 406-425
  35. Sinchaikul S, Sookkheo B, Topanuruk S, Juan HF, Phutrakul S, and Chen ST (2002) Bioinformatics, functional genomics, and proteomics study of Bacillus sp. J Chromatogr B Analyt Technol Biomed Life Sci 771, 261-287 https://doi.org/10.1016/S1570-0232(02)00054-5
  36. Teather RM and Wood PJ (1982) Use of congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Appl Environ Microbiol 43, 777-780
  37. Thompson, JD, Gibson TJ, Plewniak F, Jeanmougin F, and Higgins DG (1997) The CLUSTAL-X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25, 4876-4882 https://doi.org/10.1093/nar/25.24.4876
  38. Weisburg WG, Barns SM, Pelletier DA, and Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173, 697-703 https://doi.org/10.1128/jb.173.2.697-703.1991