Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Antimicrobial Activity of Gluten Hydrolysate with Asp. saitoi Protease

밀 단백 효소 가수분해물의 항균활성

  • Lee, Sang-Duk (Dept. of Food Science and Technology, College of Agriculture and Life Science, Chungnam National University) ;
  • Joo, Jeong-Hyeon (Dept. of Food Science and Technology, College of Agriculture and Life Science, Chungnam National University) ;
  • Lee, Gyu-Hee (Dept. of Food Science and Technology, College of Agriculture and Life Science, Chungnam National University) ;
  • Lee, K.T. (Dept. of Food Science and Technology, College of Agriculture and Life Science, Chungnam National University) ;
  • Oh, Man-Jin (Dept. of Food Science and Technology, College of Agriculture and Life Science, Chungnam National University)
  • Published : 2003.07.01
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Abstract

This study was carried out to investigate whether peptide produced from wheat protein by enzyme hydrolysis can be used as a natural antimicrobial agent. Antimicrobial peptide was obtained from wheat protein hydrolyzed by 7 of pretense. The produced antimicrobial peptide was purified through ultrafiltration, membrane filtration and HPLC and molecular weight and amino acid sequence of the purified antimicrobial peptide were determined. Among hydrolysate produced from wheat protein by 7 of protease, antimicrobial activity was observed for the peptide obtained from Asp. saito protease. The Asp. saito protease did produce antimicrobial hydrolysate showing the highest antimicrobial activity at reaction condition of 37$^{\circ}C$ and pH 6.0, but not at reaction condition above 5$0^{\circ}C$. Wheat protein hydrolysate was fractionated by membrane filtration and showed antimicrobial activity between molecular weight 1,000~3,000. The antimicrobial activity fraction obtained by membrane filtration was separated through HPLC and showed antimicrobial activity in the peak of retention time 31.1~31.8 min. We could convince this hydrolysate as heat-stable peptide since antimicrobial activity was maintained after treated with heat for 15 min at 121$^{\circ}C$. Molecular weight of antimicrobial peptide identified by MALDI-mass was 1,633. Amino acid sequence of antimicrobial peptide was cysteine, glycine, prolin, prolin, prolin, valine, valine, alanine, alanine and arginine.

밀 단백질에 효소가수 분해할 때 생산되는 peptide의 항균활성과 천연항균제로서의 이용가능성을 검토하기 위하여 실험을 행하였다. 밀 단백질에 7종의 단백질가수분해효소를 작용시켜 생성된 가수분해물의 항균활성을 측정하고 한외여과, membrane filtration, HPLC를 이용하여 항균성 peptide를 분리 정제한 후 분자량과 아미노산 결합순서를 측정한 결과는 다음과 같다. 밀 단백질에 7종의 단백질 분해효소를 적용시켜 제조한 가수분해물중 Asp. saito protease를 적용시켜 얻어진 peptide만이 항균활성을 나타내었다. Asp. saito protease는 37$^{\circ}C$, pH 6.0에서 작용시킨 경우에 항균활성이 가장 높았으며, 5$0^{\circ}C$ 이상에서는 활성을 나타내지 않았다. 밀단백 효소가수분해물은 membrane filtration에 의하여 분자량 1,000~3,000에서 항균활성이 나타났다. Membrane filtration으로 얻어진 항균활성분획을 HPLC로 분리한 결과 retention time 31.1~31.8 min에서 항균활성을 나타내었다. 밀단백 효소가수분해물은 121$^{\circ}C$에서 15분간 가열하여도 효소활성이 유지되는 매우 안정한 화합물이었다. 항균활성분획을 MALDI-mass로 질량을 분석한 결과 1,633이었다. 항균성 peptide의 아미노산 결합순서는 cysteine, glycine, prolin, valine, valine, alanine, alanine, arginine의 순서였다.

Keywords

References

  1. Naidu AS. 2000. Natural Food Antimicrobial Systems. CRC press, New York, Washington DC. p 1-11
  2. Boman HG. 1995. Antimicrobial activity of peptide. Annu Rev Immunal 13: 61-92 https://doi.org/10.1146/annurev.iy.13.040195.000425
  3. Lehrer RI, Lichtenstein AK, Geanz T. 1993. Defensin: antimicrobial and cytoxic peptides of mammalian cells. Annu Rev Immunol 11: 105-128 https://doi.org/10.1146/annurev.iy.11.040193.000541
  4. Gennaro R, Skerlavai B, Romeo D. 1989. Purification, composition and activity of two bactenecins, antibacterial pep tides of bovine neutrophils. Insect Immunol 57: 3142-3146
  5. Elsbach P, Weiss J. 1993. Bactericidal/permeability increasing protein and host defense against gram-negative bacteria and endotoxin. Curr Opin Immunol 5: 103-107 https://doi.org/10.1016/0952-7915(93)90088-A
  6. Lee JY, Boman A, Chuanxin S, Andersonn M, Jornvall H,Mutt V, Boman H G. 1989. Antibacterial peptides from pigintestine: isolation of a mammalian cecropin. Proc Natl Acad Sci USA 86: 9159-9162 https://doi.org/10.1073/pnas.86.23.9159
  7. Bevins CL, Zasloff M. 1990. Peptide from frog skin. Annu Rev Biochem 59: 395-414 https://doi.org/10.1146/annurev.bi.59.070190.002143
  8. Simmaco M, Mignogna G, Barra D, Bossa F. 1994. Antimicrobial peptides from skin secretions of Rana esculenta. J Biol Chem 269: 11956-11961
  9. Boman G. 1995. Peptide antibiotics and their role in innateimmunity. Annu Rev Immunol 13: 61-92 https://doi.org/10.1146/annurev.iy.13.040195.000425
  10. Hoffmann JA, Hetru C. 1992. Insect defensins inducibleantibacterial peptides. Immunol Today 13: 411-415 https://doi.org/10.1016/0167-5699(92)90092-L
  11. Casteels P, Ampre C, Jacobs F, Tempst P. 1993. Functional and chemical chacterization of Hymenoptaecin, an antibacterial polypeptide that is infection inducible honeybee. J Biol Chem 268: 7044-7054
  12. Hara S, Yamakawa M. 1995. Moricin, A novel type of antibacterial peptide isolated from Silkworm, Bombyx mori. J Biol Chem 270: 29923-29927
  13. Orivel J, Rederker V, Caer JL, Krier F. 2001. Ponericins, new antibacterial and insecticidal peptide from the venom of theant pachycondvla goeldii. J Biol Chem 276: 17823-17829 https://doi.org/10.1074/jbc.M100216200
  14. Bilikova K, Wu GS, Simuth J. 2001. Isolation of a peptide fraction from honeybee royal jelly as a potential antifoulbrood factor, Apidologie 32: 275-283 https://doi.org/10.1051/apido:2001129
  15. Naidu AS. 2000. Natural Food Antimicrobial Systems. CRC press, New York, Washington DC. p 31-44
  16. Wu M, Maier, E, Benz R, Hancock REW. 1999. Mechanismof interaction of different classes of cationic antimicrobialpeptides with planar bilayers and with the cytoplasmic membrane of Escherichia coli. Biochem 38: 7235-7242 https://doi.org/10.1021/bi9826299
  17. Wu M, Hancocks REW. 1999. Interaction of the cyclic antimicrobial cationic peptide bactenicin with the outer and cytoplamic membrane. J Biol Chem 274: 29-34 https://doi.org/10.1074/jbc.274.1.29
  18. Fennema OR. 1996. Food chemistry. Marcel Deker Inc., NewYork. p 423
  19. Chiba H, Yoshikawa M. 1986. Biologically functional peptides from food protein; New opioid peptides from milkprotein. Marcel Deker Inc., New York. p 123-153
  20. Dionysius DA, Milne JM. 1997. Antibacterial peptides of bovine lactoferrin: Purification and characterization. J DairySci 80: 667-674 https://doi.org/10.3168/jds.S0022-0302(97)75985-X
  21. Tomita M, Bellamy W, Takase M, Yamauchi K, WakabashiH, Kawase K. 1991. Potent antibacterial peptides generatedby pepsin digestion of bovine lactoferrin. J Dairy Sci 74:4137-4141 https://doi.org/10.3168/jds.S0022-0302(91)78608-6
  22. Pellegrini A, Thomas U, Bramaz N, Hunziker P, Fellenberg RV. 1999. Isolation and identification of three bacterial domains in the bovine $\alpha$-lactoalbumin molecule. Biochemica et Biophysica Acta 1426: 439-448 https://doi.org/10.1016/S0304-4165(98)00165-2
  23. Isidra R, Servaas V. 1999. Identification of two distinct antibacterial domains with in the sequence of bovine $\alpha$ $S_2$ casein, Biochemica et Biophysica Acta 1428: 314-326 https://doi.org/10.1016/S0304-4165(99)00079-3
  24. Pellegrini A, Dettling C, Thomas U, Hunziker P. 2001. Isolation and characterization of four bactericidal domains inthe bovine $\beta$-lactoglobulin. Biochemica et Biophysica Acta 1526: 131-140 https://doi.org/10.1016/S0304-4165(01)00116-7
  25. Kim SY, Park PSW, Rhee KC. 1990. Functional properties of proteolytic enzyme modified soy protein isolate. J Agric Food Chem 36: 651-656
  26. Manachini PL, Fortina MG, Parini C. 1988. Enzymatic modification of vegetable protein by a crude preparation froma strain of Bacillus licheniformis. J Sci Food Agric 45: 263-266 https://doi.org/10.1002/jsfa.2740450309
  27. Chobert JM, Bertrand-Harb C, Nicolas MG. 1988. Solubilityand emulsifying properties of casein and whey Proteinsmodified enzymatically by trypsin. J Agric Food Chem 36:883-892 https://doi.org/10.1021/jf00083a002
  28. Puski G. 1975. Modification of functional properties of soyproteins by proteolytic enzyme treatment. Cereal Chem 52:655-664
  29. Yamashita M, Arai S, Matsuyama J, Gonda M, Kato H, Fujimaki M. 1970. Phenomenal survey alpha-chymotrypsinplastein synthesis. Agric Biol Chem 34: 1484-1488 https://doi.org/10.1271/bbb1961.34.1484
  30. AOAC. 2000. Official Metho of Analysis. 17th ed. Association of official analytical chemists, Washington DC.
  31. Norrell SA, Messley KE. 1997. Microbiology laboratory manual: principles and applications. Prentice-Hall, Inc., New Jersey. p 191-196
  32. Hsieh DST, Lin C, Lang ER, Catsimpoolas Rha N. 1979. Molecular-weight distribution of soybean globulin peptidesproduced by peptic hydrolysis. Cereal Chem 56: 227-231
  33. Deeslie WD, Cheryan M. 1991. Fractionation of soy protein hydrolysates using ultrafiltration membrane. J Food Sci 57: 411-413

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