Bonine과 Caprine유로부터 Glycomacropeptide 분리: 트립신가수분해물의 혈소판응집억제 효과
Seperation of glycomacropeptide from bonine and caprine milk: Effect of its tryptic hydrolysate on the inhibition of platelet aggregation
0
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
본 연구는 홀스타인, 한국재래산양 및 한우유로부터 glycomacropeptide(GMP)를 분리하였으며, 각 GMP의 trypsin 가수분해물의 혈소판응집 억제 효과를 in vitro 상에서 알아보았다. 홀스타인, 한국재래산양 및 한우의 GMP는 분자량이 모두 약 20 KDa이었으며, sialic acid 함량은 각각 36.86±2.36, 37.98±1.27 및 31.19±1.87㎍/mg이었다. 또한 모든 개체의 GMP에서 tyrosine이 검출되었다. 홀스타인, 한국재래산양 및 한우 GMP의 trypsin 가수분해물에 의한 혈소판 응집 억제율은 반응 30초에 4.02, 5.51 및 12.77%로 각각 나타나 시간이 경과할수록 감소하는 경향을 보였다. 혈소판의 현미경 관찰 결과 가수분해물 첨가 후 혈소판 수가 증가하였으나, 첨가 후 30초가 경과한 시점부터 혈소판 수가 감소하기 시작하여 120초 후에는 관찰 할 수 없었다. 본 실험 결과 bovine 및 caprine GMP의 trypsin 가수분해물에서 혈소판 응집을 억제할 수 있는 small peptide가 있는 것으로 생각되며, 향후 이러한 연구는 심근경색증 및 뇌혈전증을 예방할 수 있는 생리활성 물질로 이용될 수 있을 것이라 생각된다.
Abstract
This study was carried out to isolate and characterize the glycomacropeptide (GMP) prepared from cow's milk and Korean native goat's milk and to examine the effects of their tryptic hydrolysates on inhibition of platelet aggregation in an in vitro experiment. The GMP derived from Holstein, Korean native goat and Hanwoo migrated at 20 KDa. Sialic acid contents in skim milk of Holstein, Korean native goat and Hanwoo were 36.86±2.36, 37.98±1.27 and 31.19±1.87 ㎍/mg, respectively. Tyrosine was detected in both bovine and caprine GMP. The in vitro inhibition rate of platelet aggregation by tryptic hydrolysates of Holstein, Korean native goat and Hanwoo GMP were 4.02, 5.51 and 12.77%, respectively at reaction time 30 seconds. The inhibition of platelet aggregation by tryptic hydrolysates of bovine and caprine GMP are increased with increasing reaction time. The platelets staining revealed higher counts of platelets after the addition of GMP hydrolysates; however addition of ADP reduced the platelet count within 30 seconds and the platelets were not detected after 120 seconds. The results of this study indicate that tryptic hydrolysates of bovine and caprine GMP contain some small peptides with platelet aggregation properties. Further research on these lines may help prevent platelet aggregation related abnomalities in human.
참고문헌 (31)
- AOAC, "Official methods of analysis", Association of Official Analytical Chemists, Washington. DC, USA, ~ (1995)
- Abd El-Salam, "Characteristics and potential uses of the casein macropeptide", Int. Dairy J, 6, pp.327~341 (1996)
- Anifantakis, E. M, "Contribution to the study of thecomposition of goat's milk", Milchwissenschaft, 35, pp.617~619 (1980)
- Bouhallab, S., "Tryptic hydrolysis of caseinomacropeptide in membrane ractor: Preparation of bioactive peptides. Biotech", Lett, 14, pp.805~810 (1992)
- Bradford,M.M, "A rapid and sensitive methods for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding", Anal. Biochem, 72, pp.248~254 (1976)
- Creamer, L. K, "The distribution of glyco-κ-casein and carbohydrate-free κ-casein between large and small bovine casein micelles, and its implication in micelle structure", Biochem. Biophys. Acta, 317, pp.202~208 (1973)
- Denvendra,C, "The composition of milk of British Alpine and Anglo-Nubian goats imported into Trinidad", J. Dairy Res, 39, pp.381~385 (1972)
- Doi, H, "Minor components of reduced bovine κ-casein", J. Nutr. Sci. Vitaminol, 25, pp.95~102 (1979)
- Dziuba,J.and P.Minkiewicz, "Influence of glycosylation on micelle-stabilizing ability and biological properties of C-terminal fragment of cow's κ-casein", Int. Dairy J, 6, pp.1017~1044 (1996)
- Fandino, R. L, "Comparative study by HPLC of caseinomacropeptides from cows', ewes' and goats' milk", J. Dairy Res, 60, pp.117~121 (1993)
- Foltman,B, "General and molecular aspects of rennets. In cheese: Chemistry, Physics and Microbiology Ⅰ. 2nd ed" (1993), Chapman & Hill.
- Hawiger,J, "Platelets: receptors, adhesion, secretion" (1989), Academic Press
- Jolles,P, "Structural aspects of the milk clotting process. Comparative features with the blood clotting process", Mol. Cell. Biochem, 7, pp.73~85 (1975)
- Jolles, P, "Comparison between the clotting of blood and milk", Trends Biochem. Sci, 7, pp.325~328 (1982)
- Jolles, P, "Analogy between fibrinogen and casein: Effect of an undecapeptide isolated from κ-casein on platelet function", Eur. J. Biochem, 158, pp.379~382 (1986)
- Kawasaki, Y, "Inhibition by κ-caseinoglycomacropeptide and lactoferrin of influenza virus hemagglutination. Biosci", Biotech. Biochem, 57, pp.1214~1215 (1993)
- Kim, S. B, "Enzymatic Hydrolysis of Heated Whey: Iron-Binding Ability of Peptides and Antigenic Protein Fractions", J. Dairy Sci, 90, pp.4033~4042 (2007)
- Korhonen, H, "Bioactive peptides: Production and Functionality", Int. Dairy J., 16, pp.1262~1271 (2006)
- Laemmli,U.K, "Cleavage of structural proteins during the assembly of the head of bacteriophage T4", Nature (Lord)., 227, pp.680~685 (1970)
- Leonil, J, "A method for determination of macropeptide of cation-exchange fast protein liquid chromatography and its use for following the action of chymosin in milk.J.Dairy Res.58:321-328", J. Dairy Res, 58, pp.321~328 (1991)
- Longenecker,G.L, "The platelets: physiology and pharmacology", pp.1~14 (1985), Academic Press, Orlando
- Mazoyer, E, "KRDS, a new peptide derived from human lactotransferrin, inhibits platelet aggre-gation and release reaction", Eur. J. Biochem, 194, pp.43~49 (1990)
- Moore, S, "Automatic recording apparatus for use in the chromatography of amino acids.Fed Proc.17:1107-1115", Fed Proc, 17, pp.1107~1115 (1958)
- Morr, C. V, "Fractionation and characterization of glycomacropeptide from caseinate and skim milk hyrolysates", J. Food Sci, 53, pp.80~87 (1988)
- O'Conner, P, "The proteins and salts of some non-bovine milks", J. Dairy Res, 44, pp.607~609 (1977)
- Saito, T, "A new isolation method ofcaseinoglycopeptide from sweet cheese whey", J. Dairy Sci, 74, pp.2831~2837 (1991)
- Tanimoto, M, "Largescale preparation of κ-casein glycomacropeptide from rennet casein whey", Biosci. Biotechnol. Biochem, 56, pp.140~141 (1992)
- Warren,L, "The thiobarbituric acid assay of sialic acids", J. Biol. Chem, 234, pp.1971~1975 (1959)
- 김의수, "카제인으로부터 고정화 카이모신을 이용한 CMP 생산", 석사, pp.1997
- 성내경, "SAS", STAT-분산분석, 1997
- 오세종, "κ-casein에서분해된 Glycomacropeptide의 분리및 생리활성에 관한연구", 박사, pp.1996
