Purification and Partial Amino Acid Sequence of a Bacteriocin Produced by Lactococcus, sp. HY449

Lactococcus sp. HY449가 생산한 Bacterisocin의 정제

  • 오세종 ((주)한국야쿠르트 중앙연구소) ;
  • 이상준 (청강문화산업대학 건강식품과학과) ;
  • 김경태 ((주)한국야쿠르트 중앙연구소) ;
  • 김상교 ((주)한국야쿠르트 중앙연구소) ;
  • 박연희 (아주대학교 생물공학과) ;
  • 백영진 ((주)한국야쿠르트 중앙연구소)
  • Published : 2001.09.01

Abstract

A bactriocin produced by Lactrococcus sp. HY449 was purified by sequential purfication steps such as n-propanol-acetone precipitation ion -exchange chromatography using CM-Sequential CL6B. gel filtration chromatography using Sephacry HR100 and reverse-phase chromatography using pro RPC HR 5/10. Reverse-phase chromatography the final step of the purfication yielded a single symmetrical peak of bacteriocin activity The purification resulted in final yield of 3.25% and 413.35 fold increase of the specific activity of bacteriocin. The active fraction from reverse-phase chromatography was used for N-terminal amino acid analysis . The purified bacteriocin contained isoleucine, leucine, methionine, and glycine at but N-terminal end no aromatic amino acids. Calculation of the number of amino acid residues in the bacteriocin revealed that it is consisted of 32 residues assuming the molecular weight of bacteriocin to be about 3.6kDa. Edman degrandation elucidated amino acid residues of the first four of the N-terminus to be $NH_2$-Ile-Leu-Pro-GIn.

Keywords

Bacteriocin;Lactococcus sp;HY449;purification;chromatography

References

  1. Kor. J. Appl. Microbiol. Biotchnol. v.22 Optimizing conditions for the growth and bacteriocin production of Lactococcus sp. HY 449 using response surface methodology Kim, S. K.;S. J. Oh;S. J. Lee;Y. J. Baek;Y. H. Park
  2. Appl. Environ. Microbiol. v.59 Isolation and purifiaction of propionicin PLG-1, a bacteriocin produced by a strain of Propionibacterium thoenii Lyon, W. J.;B. A. Glatz
  3. Appl. Environ. Microbiol v.57 Purification amd partial characterization of Lactacin F, a bacteriocin produced by Lactobacillus acidophilus11088 Muriana, P. M.;T. R. Klaenhammer
  4. Appl. Environ. Microbiol. v.58 Purification and partial characterization of lacticin 481, a lanthionine-containing bacteriocin produced by Lactococcus lactis subsp. lactis CNRZ 481 Piard, J. C.;P. M. Muriana;M. J. Desmazeaud;T. R. Klaenhammer
  5. Biochem. Biophys. Res. Commum. v.35 High recovery of tryptophan from acid hydrolysates of proteins Matsubara, H.;R. M. Sasaki
  6. J. Food Sci. v.44 Antimicrobial activity of lactic cultures: partial purification and characterization of antimicrobial compound(s) produced by Streptococcus thermophilus Pulusani, S. R.;K. R. Rao;G. R. Sunki
  7. J. Bacteriol. v.167 Characterization and purification of helveticin J and evidence for a chromosomally determinced bacteriocin produced by Lactobacillus helveticus 418 Joerger, M. C.;T. R. Klaenhammer
  8. Food Rev. Intl. v.9 Bacteriocins of lactic acid bacteria: Their potentials as food biopreservative Kim, W. J.
  9. Bacteriocins of lactoic acid bacteria Antimicrobial proteins: classification, nomenclature, diversity, and relationship to bacteriocins Montville, T. J;A. L. Kaiser;D. G. Hoover;L. R. Steenson(ed.)
  10. Food Technol. v.44 Nisin and its uses as a food preservative Delves-Broughton, J.
  11. Anal. Biochem. v.141 Sodium sulfate as an antioxidant in the acid hydrolysis of bovine pancreatic ribonuclease A Swadesh, J. K.;T. W. Thannhauser;H.A. Scheraga
  12. Appl. Environ. Mrcrobiol. v.57 Purification and amino acid sequence of lactocin S, a bacteriocin produced by Lactobacillus sake L45 Mortvedt, C. I.;J. Missen-Meyer;K. Sletten;I. F. Nes
  13. Appl. Environ. Microbiol. v.58 Novel method to extract large amounts of bacteriocin from lactic acid bacteria Yang, R.;M.C. Johnson;B. Rayvel
  14. Kor. J. Appl. Microbiol. Biotechnol. v.22 Isolation fo bacteriocin-producing Lactococcus sp. HY 449 and its antimicrobial characteristics Kim, S. K.;S. J. Lee;Y. J. Baek;Y. H. Park
  15. Anal. Biochem. v.166 Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa Sch$\"{a}$gger, H.;G. V. Jagow
  16. J. Dairy Sci. v.83 Characterization and purification of a bacteriocin produced by Lactobacillus acidophilus 30SC: Human isolate for potential use as a probiotic strains Oh, S.;S. Kim;R. W. Worobo
  17. J. Kor. Dairy Technol. & Sci. v.19 Characteristics of class ∥ bacteriocins produced by lactic acid bacteria Oh, S.
  18. FEMS Microbiol. Rev. v.12 Genetics of bacteriocins produced by lactic acid bacteria Klaemhammer,T. R.
  19. Bacteriol. Rev. v.40 Bacteriocins of gram-positive bacteria Tagg, J. R.;A. S. Dajani;L. W. Wannamaker
  20. Food Technol. v.43 Antimicrobial substances from lactic acid bacteria for use as food preservatives Daeschel, M. A.
  21. Kor. J. appl. Microbial. Biotechnol. v.19 Purification and properties of bacteriocin produced by Lactococcus sp. 1112-1 Choi, S. Y.;S. H. Lee;I. S. Lee;J. Y. Yoo;K. S. Chung;Y. J. Koo
  22. J. Appl. Bactreiol. v.69 Antibacterial activity of lactic acid bacteria isolated from vacuum-packaged meats Ahn, C.;M. E. Stiles