Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Isolation and Characterization of Chitinoloytic Strain, Bacillus atrophaeus CJ-3.

전통발효식품으로부터 Chitin 분해 미생물의 분리 및 특성 규명

  • 고보경 (신라대학교 식품영양학과) ;
  • 최인순 (신라대학교 생물과학과) ;
  • 이상현 (신라대학교 생명공학과) ;
  • 임채오 (경상대학교 응용생명과학부) ;
  • 이성호 (경상대학교 응용생명과학부) ;
  • 갈상완 (진주산업대학교 미생물공학과) ;
  • 최영주 (신라대학교 식품영양학과)
  • Published : 2004.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

A bacterial strain CJ-3 which produced chitinase was isolated from Korean traditional soy sauce. Using 16S rDNA analysis, the strain CJ-3 was identified as Bacillus atrophaeus. The approximate molecular weight of the putative chitinase enzyme was 31.0 kDa and the enzyme activity was remarkably induced by addition of colloidal chitin (0.5, 1.0, 2.0%). The antioxidant activity was increased 53% by the browning reaction products of B. atrophneus CJ-3. Escherichia. coli lipopolysaccharides (LPS)-induced production of nitric oxide(NO) was reduced up to 45% by the browning reaction product in RAW264.7 macrophage. Inhibition of cell viability in the presence of LPS was recovered to normal level by the browning reaction product. These results suggest that browning reaction of B. atrophaeus CJ-3 plays an important role for activation of immune system. B. atrophaeus CJ-3 exhibited optimum temperature and pH of 37$^{\circ}C$ and pH 7.0∼8.0, respectively. The major intracelluar free amino acid was determined to be glutamate.

전통발효 식품인 간장으로부터 chitinase활성이 높으면서 갈변반응을 일으키는 균주인 CJ-3를 분리하여 갈변반응생성물이 면역기능, 항산화력 및 세포활성에 미치는 영향과 생화학적 특성을 규명하였다. 분리된 CJ-3 균주는 16S rDNA 염기서열분석에 의하여 Bacillus atrophaeus로 동정되었으며 분리된 균주의 분자량은 대략 31.0 kDa으로 colloidal chitin(0.5, 1.0, 2.0%)의 첨가에 의하여 chitinase 활성이 현저히 증가되었다 B. atrophaeus CJ-3에 의해서 유도되는 갈변반응물 200${\mu}\ell$ 첨가로 LPS에 의하여 유도되는 nitric oxide (NO) 활성을 45%까지 감소시켰다. MTT 분석을 통한 세포활성에서도 갈변반응물은 LPS에 의하여 세포활성을 정상수준으로 회복하였다. CJ-3. 균주의 DPPH 전자공여능법에 의한 항산화력은 갈변반응에 의하여 약 55% 증가하였다 . 이러한 결과들은 갈변반응생성물은 면역기능 및 세포활성을 증가시키는 것으로 추정된다. 분리된 균주의 최적 생육조건은 최적온도 37$^{\circ}C$, 최적 pH 7.0 및 염 농도는 9% NaCl농도까지는 잘 생육하는 것으로 나타났으며 분리된 균주의 주요 intracellular 유리아미노산은 glutamate로 나타났다.

Keywords

References

  1. Annual report on the cause of death statistics National Statitical Office
  2. Kor. J. Food Sci. Technol. v.31 Isoflavone contents and β-glucosidase activities of soybeans, Meju and Doenjang Kim,J.S.;S.Yoon
  3. J. Agric. Food Chem. v.47 Estrogenic activity of glycitein, a soy isoflavone Song,T.T.;S.Handrich;P.A.Murphy https://doi.org/10.1021/jf981054j
  4. Life Sci. v.73 Screening of medicinal extracts for antioxidant activity Lee,S.E.;H.J.Hwang;J.S.Ha;H.S.Jeong;J.H.Kim https://doi.org/10.1016/S0024-3205(03)00259-5
  5. J. Agric. Food Chem. v.48 Antioxidant properties of aroma compounds isolated from soybeans and mung beans Lee,K.G.;T.Sjibamoto https://doi.org/10.1021/jf000442u
  6. Kor. J. Food Sci. Technol. v.22 Antioxidative characteristics of fermented soybean sauce on the oxidation of fatty acid mixture Cheigh,H.S.;J.S.Lee;G.S.Moon;K.Y.Park
  7. FEBS Letters v.556 Overexpression of redox factor-1 negatively regulates NO synthesis and apoptosis in LPS-stimulated RAW 264.7 macrophages Yoo,Y.H.;Y.J.Lim;S.E.Park;J.M.Kim;Y.C.Park https://doi.org/10.1016/S0014-5793(03)01361-9
  8. Adv. Food Res. v.32 Chemistry of Maillard reactions : recent studies on the browning reaction mechanism and the development of antioxidants and mutagens Namiki,M. https://doi.org/10.1016/S0065-2628(08)60287-6
  9. J. Korea Agric. Chem. Soc. v.31 Desmutagenicity of the enzymatic browning reaction products which obtained from Prunus salficiana (yellow) enzyme and polyphenol compounds Ham,S.S.
  10. Prog. Food Nutrition Sci. v.5 Antioxidative effect of Maillard reaction intermediates Eichner,K.
  11. J. Agric. Food Chem. v.50 Inhibitory effect of natto, a kind of fermented soybeans, on the LDL oxidation in vitro Iwai,K.;N.Nakaya;Y.Kawasaki;H.Matsue https://doi.org/10.1021/jf011718g
  12. Biosci. Biotechnol. Biochem. v.75 Purification and characterization of the antioxidative substance produced by Aspergillus sojae Rashid,M.H.O.;F.Kato;A.Murata;M.Kondo
  13. Agric. Food Chem. v.50 Antioxidative activity and safety of the 50 ethanol extract from red bean fermented by Bacillus subtilis IMR-NKI Chung,Y.C.;C.T.Chang;W.W.Chao;C.F.Lin;S.T.Chou https://doi.org/10.1021/jf011369q
  14. Kor. Soc. Agric. Chem. Biotechnol. v.46 Antioxidative and produced condition of antioxidative substance by Bacillus sp. FF-7 Cha,J.Y.;H.J.Kim;B.S.Jun;J.C.Park;M.Ok;Y.S.Cho
  15. J. Kor. Soc. Food Sci. Nutr. v.25 Characteristics and action pattern of protease from Bacillus subtilis CCKS-111 in Korean traditional soy sauce Choi,C.;K.S.Choi;Y.J.Cho;S.I.Lim;S.Kim;J.H.Son;H.D.Lee;Y.H.Kim
  16. Kor. J. Chitin and Chitin-san v.1 Bioactivities of chitin and chitinsan (I) Jeon,Y.J.;E.H.Lee;S.K.Kim
  17. Kor. J. Chitin and Chitin-san v.1 Synthetic techniques of chitin and chitin-san derivatives and their analytical methods Kim,S.K.
  18. J. Gen. Microbiol. v.134 Plant and bacterial chitinases differ in antifungal activity Roberts,W.K.;C.P.Selitrennikoff
  19. J. Mol. Biol. v.215 Basic local alignment search tool Altschul,S.F.;W.Miller;E.W.Myers;D.J.Lipman https://doi.org/10.1016/S0022-2836(05)80360-2
  20. Nucleic. Acids Res. v.21 The ribosomal database project Larsen,N.;G.J.Olsen;B.L.Maidak;M.J.McCaughey;R.Overbeek;T.Macke;T.L.Marsh;C.Woese https://doi.org/10.1093/nar/21.13.3021
  21. Anal. Biochem. v.72 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Bradford,M.M. https://doi.org/10.1016/0003-2697(76)90527-3
  22. Nature v.26 Antioxidant determination by the use of a stable free radical Blois,M.S.
  23. Mol. Gen. Genet. v.182 Proline over-production results in enhanced osmotolerance in Salmonella typhimurium Csonka,L.N. https://doi.org/10.1007/BF00422771
  24. Appl. Environ. Microbiol. v.40 Diversity and dynamics of indigenous Rhizobium japonicum population Noel,K.D.;W.J.Brill
  25. Nature v.227 Cleavage of structure protein during the assembly of head of bacteriophage T4 Laemmli,U.K. https://doi.org/10.1038/227680a0
  26. Biochem. Pharmacol. v.66 Chen. Inhibition of lipopolysaccharide-induced nitric oxide production by flavonoids in RAW264.7 mqcrophages in volves heme oxygenase-1 Lin,H.Y.;S.H.Juan;S.C.Shen;F.L.Hsu;Y.C. https://doi.org/10.1016/S0006-2952(03)00422-2
  27. Kor. J. Appl. Microbiol. Biotechnol v.21 Identification and cultural characterization of Streptomyces lydicus G-23 for producing chitinase Lee,S.M.
  28. J. Ko.n Soc. Agric. Chem. Biotechnol. v.42 Isolation and antifungal activity of the chitinase producing bacterium Serratia sp 3095 as antagonistic bacterium against Fusarum sp. Lee,E.T.;S.D.Kim
  29. Kor. J. Food Sci. Technol. v.7 Comparison of the antioxidant effects of ethyl alcohol extracts of a maillard-type and a caramelization-type browning reaction mixtures Lee,D.I.;T.R.Heo;D.H.Kim
  30. Kor. J. Food Sci. Technol. v.29 Effect of maillard reaction products on growth of Bacillus sp Lee,G.D.;J.S.Kim;J.H.Kwon
  31. Kor. J. Appl. Microbiol. Biotechnol. v.24 Purification and characterization of thermostable chitinase from Bacillus licheniformis KFB-V14 Hong,B.S.;H.G.Yoon;D.H.Shin;H.Y.Cho
  32. Kor. J. Life Sci. v.12 Purification and characterization of the chitinase from Bacillus subitilits JK-56 Kim,N.W.;Y.K.Jeong;H.K.Jun https://doi.org/10.5352/JLS.2002.12.1.077
  33. Current Microbiol. v.39 Purification and characterization of chitinase from Bacillus circulans No. 4.1 Wiwat,C.;P.Siwayaprahm;A.Bhumiratana https://doi.org/10.1007/s002849900434
  34. J. Biol. Chem. v.268 Nitric oxide synthase structure and mechanism Marletta,M.A.
  35. FASEB J. v.6 Nitric oxide as a secretary product of mammalian cells Nathan,C.
  36. Arch. Biochem. Biophys v.362 Snitrosylation and S-glutathiolation of protein sulfhydryls by S-nitroso glutathione Ji,Y.;T.P.Akerboom,H.Sies;J.A.Thomas

Cited by

  1. Isolation and Characterization of Bacillus velezensis SS360-1 from Seed Soy Sauce vol.29, pp.1, 2018, https://doi.org/10.7856/kjcls.2018.29.1.49