Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Identification of the Cell-envelope Proteinase of Lactic Acid Bacteria Isolated from Kimchi.

김치 유래 젖산균의 Cell-envelope Proteinase 존재 확인

  • 이유진 (경기대학교 식품생물공학과) ;
  • 최재연 (경기대학교 식품생물공학과) ;
  • 이형주 (서울대학교 식품공학과) ;
  • 장해춘 (조선대학교 식품영양학과) ;
  • 김정환 (경상대학교 식품공학과) ;
  • 정대균 (경희대학교 유전공학과 및 유전공학연구소) ;
  • 김영석 (이화여자대학교 식품영양학과) ;
  • 김소미 (제주대학교 아열대 원예산업 연구센터) ;
  • 이종훈 (경기대학교 식품생물공학과)
  • Published : 2002.06.01
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Abstract

The partial 165 rDNA sequences of 6 lactic acid bacterial strains isolated from Kimchi were determined. Two strains were Leuconostoc mesenteroides and the rest were incorrectly classified and turned out to be Lactobacillus. As the case of dairy lactic acid bacteria, the strains isolated from Kimchi also had cell-envelope proteinase (CEP) activity. As the result of partial CEP gene amplification with CEP-specific primers, the expected 1.2-kb amplificate was obtained not from Leu. mesenteroides but from Lactobacillus strains. The deduced amino acid sequence of PCR product amplified from the genomic DNA of Lactobacillus pentosus KFR1821 showed 95％ and 92％ homology with those of PrtPs from Lactococcus lactis subsp. cremoris and Lactobacillus paracasei subsp. paracasei, respectively. The PCR amplificate was used as a probe and the result of Southern hybridization illuminated the location of CEP gene in chromosomal DNA of Lb. pentosus KFR1821.

김치로부터 분리된 젖산균들의 16S rDNA 부분 염기서열 결정 결과, 6균주 중 2균주는 Leuconostoc mesenteroides로, 나머지 균주는 Lactobacillus속으로 재동정되었다. 이들 김치유래 젖산균들은 유제품 유래 젖산균과 마찬가지로 cell-envelope proteinase (CEP)로 추정되는 세포외 proteinase 활성을 보유하고 있는 것으로 나타났고, 젖산균의 CEP 특이적 primer를 제작하여 PCR을 수행한 결과, Leu. mesenteroides로부터는 예상 크기의 PCR 산물이 증폭되지 않았지만 Lactobacillus속 균주들로부터는 예상되는 1.2 kb 의 DNA 단편이 증폭되었다. Lactobacillus pentosus KFR1821의 genomic DNA로부터 증폭된 PCR 산물의 추정 아미노산서열은 Lactococcus lactis subsp. cremoris의 PrtP와 95％, Labtobacillus paracasei subsp. paracasei이 PrtP와 92％의 높은 상동성을 보였다. 증폭된 PCR 산물을 probe로 하여 Southern hybridization을 수행한 결과, Lb. pentosus KFR1821의 CEP 유전자는 chromosomal DNA에 암호화 되어있는 것으로 확인되었다.

Keywords

References

  1. Appl. Environ. Microbiol v.46 Simple and rapid method for isolating large plasmid DNA from lactic streptococci Anderson, D. G.;L. L. McKay
  2. Neth. Milk Dairy J. v.30 Comparison of strains of Streptococcus cremoris for proteolytic activities associated with the cell wall Exterkate, F. A.
  3. Appl. Environ. Microbiol v.47 Location of peptidases outside and inside the membrane of Streptococcus cremoris Exterkater, F. A.
  4. J. Gen. Microbiol v.138 Cloning and partial sequencing of the proteinase gene complex from Lactococcus lactis subsp. lactis UC317 Fitzgerald, G.;J. Law;P. Vos;F. Hayes;G. Daly;W. M. de Vos https://doi.org/10.1099/00221287-138-4-709
  5. J. Bacteriol v.178 A new cell surface proteinase: sequencing and analysis of the prtB gene from Lactobacillus delbrueckii subsp. bularicus Gilbert, C.;D. Atlan;B. Blance;R. Portalier;J. E. Germond;L. Lapierre;B. Mollet https://doi.org/10.1128/jb.178.11.3059-3065.1996
  6. Kor. J. Food Sci. Technol. v.22 Determination of microbial community as an indicator of Kimchi fermentation Han, H. U.;C. R. Lim;H. K. Park
  7. J. Gen. Microbiol. v.131 Cloning, sequencing and expression of the gene encoding the cell-envelope-associated proteinase from Lactobacillus paracasei NCDO151 Holck, A.;H. Naes
  8. J. Dairy Sci. v.75 Characterization of Leuconostoc isolated from commercial mixed strain mesophilic starter cultures Johansen, E.;A. Kibenich https://doi.org/10.3168/jds.S0022-0302(92)77865-5
  9. J. Bacteriol v.177 The extracellular PI-type proteinase of Lactococcus lactis hydrolyzes β-casein into more than one hundred different oligopeptides Juillard, V. H.;Laan, E. R. S. Kunji;C. M. Jeronimus-Stratingh;A. P. Bruins;W. N. Konings https://doi.org/10.1128/jb.177.12.3472-3478.1995
  10. Mol. Microbiol. v.3 Molecular characterization of a cell wall-associated proteinase gene from Streptococcus lactis NCDO763 Kok, J.;K. J. Leenhouts;A. J. Haandrikman;A. M. Ledeboer;G. Venema https://doi.org/10.1111/j.1365-2958.1989.tb00181.x
  11. Appl. Environ. Microbiol v.54 Nucleotide sequence of the cell wall proteinase gene of Struptococcus cremoris Wg2 Kok, J.;K. J. Leenhouts;A. J. Haandrikman;A. M. Ledeboer;G. Venema
  12. Appl. Environ. Microbiol v.55 Mechanism of proteinase release from Lactococcus lactis subs. cremoris Wg2 Laan, H.;W. N. Konigs
  13. Appl. Microbiol. Biotechnol. v.36 Cell-wall associated proteinase of Lactobacillus delbrueckii subsp. bulgaricus CNRZ397: differential extraction, purification and properties of the enzyme Laloi, P.;D. Atlan;B. Blance;C. Gilbert;R. Portalier https://doi.org/10.1007/BF00164419
  14. Kor. J. Appl. Microbiol. Biotechnol v.27 Changes of lactic acid bacteria and selective inhibitory substances against homo and hetro lactic acid bacteria isolated from Kimchi Lee, S. H.;N. Y. Park;W. J. Choi
  15. Appl. Microbiol. Biotechnol v.40 Purification and characterization of the mature, membrane associated cell-envelope proteinase of Lactobacillus helveticus L89 Martin-Hemandez, C.;A. Alting;F. Exterkate https://doi.org/10.1007/BF00173983
  16. Nucleic Acids Res. v.18 Nucleotide sequence of 16S ribosomal RNA from Lactobacillus viridescens and Lactobacillus confuses Martinez-Murcia, A. J.;M. D. Collins https://doi.org/10.1093/nar/18.11.3402
  17. FEMS Microbiol. Lett. v.36 Specificity of a cell wall proteinase from Streptococcus lactis NCDO763 towards bovine β-casein Monnet, V.;D. Lebars;J. C. Gripon
  18. Appl. Microbiol. Biotechnol v.31 Comparison of cell wall proteinases from Lactococcus lactis subsp. cremoris ACI and Lactococcus lactis subsp. lactis NCDO763 Monnet, V.;W. Bockelman;J. C. Gripon;M. Teuber https://doi.org/10.1007/BF00262446
  19. J. Gen. Microbiol. v.138 Purfication and N-terminal amino acid sequence determination of the cell-wall-bound proteinase from Lactobacillus paracasei subsp. paracasei Naes, H.;J. Nissen-Meyer https://doi.org/10.1099/00221287-138-2-313
  20. J. Bacteriol. v.181 Genetic characterization of a cell envelope-associated proteinase from Lactobacillus helveticus CNRZ32 Pederson, J. A.;G. J. Miliski;B. C. Weimer;J. L. Steele
  21. FEMS Microbiol. Rev. v.12 The physiology and biochemistry of the proteolytic system in lactic acid bacteria Pritchard, G. G.;T. Coolbear https://doi.org/10.1111/j.1574-6976.1993.tb00018.x
  22. Antonie van Leeuwenhoek v.76 Multi-domain, cell-envelope proteinases of lactic acid bacteria Roland, J. S. https://doi.org/10.1023/A:1002036906922
  23. Appl. Environ. Microbiol. v.66 Streptococcus thermophilus cell wall-anchored proteinase: release, purification, and biochemical and genetic characterization Rul, R.;D. R. Maria;P. Garault;B. Monnet https://doi.org/10.1128/AEM.66.11.4772-4778.2000
  24. Kor. J. Food Sci. Technol v.28 Changes of chemical composition and microflora in commercial Kimchi Shin, D. H.;M. S. Kim;J. S. Han;D. K. Lim;W. S. Bak
  25. Protein Sci. v.6 Subtilisin-like serine proteases Siezen, R. J.;W. M. de Vos;J. A. Leunissen;B. W. Dijkstra https://doi.org/10.1002/pro.5560060301
  26. Protein Eng. v.4 Homology modelling and protein engineering strategy of subtilases, the family of subtilisin-like serine proteinases Siezen, R. J.;W. M de Vos;J. A. Leunissen;B. W. Dijkstra https://doi.org/10.1093/protein/4.7.719
  27. Appl. Environ. Microbiol v.57 Casein utilization by lactococci Smid, E. J.;B. Poolman;W. N. Konings
  28. Kor. J. Food Sci. Technol. v.27 Identification of psychrotrophic lactic acid bacteria isolated from Kimchi So, M. H.;Y. B. Kim
  29. J. Mol. Biol. v.98 Detection of specific sequences among DNA fragments separated by gel electrophoresis Southern, E. M. https://doi.org/10.1016/S0022-2836(75)80083-0
  30. J. Dairy Res. v.60 Proteolytic enzymes of Lactococcus lactis Tan, P. S. T.;B. Poolman;W. N. Konings https://doi.org/10.1017/S0022029900027606
  31. Net. Milk Dairy J. v.35 Proteolytic enzymes of starter bacteria Thomas, T. D.;O. E. Mills
  32. Appl. Environ. Microbiol v.65 Cell-wall-bound proteinase of Lactobacillus delbrueckii subsp. lactis ACA-DC178: characterization and specificity for β-casein Tsakalidou, E.;R. Anastasiou;I. Vandenberghe;J. van Beeumen;G. Kalanizopoulos
  33. Appl. Environ. Microbiol. v.61 Oligopeptides are the main source of nitrogen for Lactococcus lactis during growth in milk Vincent, J.;D. L. Bars;E. R. S. Kunji;W. N. Konings;J. C. Gripon;J. Richard
  34. J. Bacteriol v.171 A maturation protein is essential for production of active forms of Lactococcus lactis SK11 serine proteinase located in or secreted from the cell envelope Vos, P.;M. van Asseldonk;F. van Jeveren;R. Siezen;G. Simons;W. M. de Vos https://doi.org/10.1128/jb.171.5.2795-2802.1989
  35. Lett. Appl. Microbiol. v.20 Detection of dairy Leuconostoc strains using the polymerase chain reaction Ward, L. J. H.;J. C. S. Browon;G. P. Davey https://doi.org/10.1111/j.1472-765X.1995.tb00428.x
  36. Sys. Appl. Microbiol. v.12 Phylogenetic structure of the Leuconostoc: an interesting case of a rapidly evolving organism Yang, D.;C. R. Woese https://doi.org/10.1016/S0723-2020(89)80005-0
  37. Lait v.68 Properties and specificity of a cell-wall proteinase from Lactobacillus helveticus Zevaco, C.;J. C. Gripon https://doi.org/10.1051/lait:1988425