산양유 단백질 가수분해물로부터 Angiotensin Converting Enzyme저해 Peptides의 분리 및 정제
Separation and Purification of Angiotensin Converting Enzyme Inhibitory Peptides Derived from Goat's Milk Proteins Hydrolysates
산양유 단백질 가수분해물 응용생명과학;
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To investigate the basic information and the possibility of ACE inhibitory peptides for antihypertension materials, goat's proteins(casein and whey) were hydrolyzed by various proteolytic enzymes(protease S, neutrase 0.8, trypsin, pepsin, and papain W-40) and ACE inhibitory peptides were separated and purified from enzymatic hydrolysates of goat's casein and whey. The general composition of goat's milk, degree of hydrolysis(DH), ACE inhibition ratios of enzymatic hydrolysates of goat's casein and whey, amino acid composition and amino acid sequence of ACE inhibitory peptides obtained by Sephadex G-25 gel chromatograms and RP-HPLC were determinated. And also IC_(50) of peptic hydrolysates and fractions separated and purified by Sephadex G-25 gel chromatograms and RP-HPLC with the highest ACE inhibition activity were determinated. 1. Urea-PAGE of goat's casein hydrolysates by pepsin were shown that all casein were converted to small peptides below 14.2 KDa after 8 hours. By SDS-PAGE of goat's whey hydrolysates by pepsin, BSA and α-lactalbumin were almost disappeared at 0.25 hours but β-lactoglobulin was slowly hydrolyzed from 8 hours. 2. The protease S(60.8 %) and trypsin(34.3 %) were the highest degree of hydrolysis among various enzymatic hydrolysates of goat's casein and whey, respectively. 3. ACE inhibition ratios of goat's casein and whey hydrolysates were shown the highest with 87.84 % by pepsin for 48 hours and with 85.50 % by pepsin for 72 hours, respectively. 4. By Sephadex G-25 gel chromatograms, fraction CF3 from goat's casein hydrolysates by pepsin for 48 hours and fraction WF4 from goat's whey hydrolysates by pepsin for 72 hours were confirmed the highest ACE inhibition activity. 5. Fraction CF3g and CF3gh from casein hydrolysates and fraction WF4e and WF4ee from whey hydrolysates by RP-HPLC to first and secound purification were the highest in ACE inhibition activity, respectively. 6. The most abundant amino acid was leucine 18.83 % in fraction CF3gh and 18.58 % in fraction WF4ee of ACE inhibitory peptides after secound purification 7. Amino acid sequence analysis of fraction CF3gh and WF4ee of ACE inhibitory peptides were shown that the Ala-Tyr-Phe-Tyr, Pro-Tyr-Tyr, Tyr-Leu and Leu-Lys-Asp-Tyr-Gly-Gly-Val-Ser-Leu, Leu-Gly-Asp-Gly-Ala-Gly-Asp-Val-Ala-Phe, respectively. 8. IC_(50) calibrated in peptic hydrolysates at 48hrs, fraction CF3, CF3g and CF3gh from goat's casein hydrolysates by pepsin for 48 hours were 29.89, 3.07, 1.85 and 0.87 ㎍/ml, respectively and those of peptic hydrolysates at 72hrs, fraction WF4, WF4e and WF4ee from goat's whey hydrolysates by pepsin for 72 hours were 33.93, 28.35, 11.74 and 1.09 ㎍/ml, respectively.