,1297272 acidophUus、L. !actis、L re齡i、L brevis)、縫球菌屬 (Streptococcus s阶乂例如 Strept〇c〇ccus therm〇pM_、雙又 乳酸桿菌屬卿祕⑽spp )(例如师祕她n._ hngum、B. bifidmt、b. in細tis、β b謂、B hctis傳但 並不僅限於此。應用於本發明所指出之方法中進行發酵的 乳酸菌’可以使用單一乳酸菌株，或混以多種乳酸菌株同 犄進订發酵’上述此兩種方法皆常被應用於習知發酵乳的 φ 製備上，因此於本發明中並沒以特別的限制。 #轉素水解方式製備時，可被應祕本發明中的酵 t，在此亦無制的_，任何f知或未知可被應用於水 解蛋白質的酵素皆可被應用於本發明中，在此可舉出的例 子包含Prozyme 6®、Flav贿yme®、Pr〇t_妒等但並 不僅限於此。前述酵素的施用濃度較佳為0.1〜1.0% 相對於基質的蛋白質總量），更佳為G.3〜〇.7% (w/w)。 嗾習本發明技術領域之技藝者亦可藉由閱讀本說明書 # 崎知，本發明寡胜肽亦可藉由習知之胜肽合成法以人工 方式合成，或藉㈣知之紐合成儀而輕易獲得。 …本發轉紐對血管升齡轉換義屬於競爭型抑制 $態。經以體外酵素抑制職後，可證實本發明寡胜敗確 實可抑制血管升壓素轉換酶之作用。 實施例一 、+、配衣緩衝，谷液A(l〇〇mM硼酸緩衝溶液，ρΗ8·3)及緩衝 合液B(3 600m]y[氯化納之1〇〇mM石朋酸緩衝溶液，ρΗ8 3)。 ‘1297272 配製jk管升壓素轉換酶(ACE)溶液··將5單位(Unit)的 血管升壓素轉換酶粉末，加入5〇ml的緩衝溶液A中，使 用時加入等量的緩衝溶液B，以製得5〇mU/ml之血管升壓 素轉換酶溶液。 馬尿酸-組胺酸-白胺酸(Hippuryl-His-Leu，簡稱HHL) /谷液之配I ·將500mg HHL粉末，各加入38.8ml之緩衝 溶液A及B，混合均勻可得15mM之HHL溶液。 另外’以胜肽合成儀核成本發明之寡胜肽。於測定時， 將本發明寡胜肽溶於5ml的緩衝溶液A與5ml的緩衝溶液 B之此合浴液中。之後，自其中取出75瓜，並與acE溶 液75μί混合，於37°C下預熱1〇分鐘，再加入15mM之 HHL溶液為基質。混合液於3rt水浴反應3〇分鐘後，再 加入0.25ml的1N鹽酸終止反應。生成的馬尿酸以高效能 液相層析法(HPLC)法，於228nm下偵測其吸光值，以測定 抑制ACE活性的百分比。 計算公式如下：抑制能力(％)=【(Ac_As)/(Ac_Ab)】 χΐ〇〇, 1297272 acidophUus, L. !actis, L re age i, L brevis), Staphylococcus (Streptococcus s order, such as Strept〇c〇ccus therm〇pM_, double Lactobacillus genus (10) spp) (eg teacher her N._ hngum, B. bifidmt, b. in fine tis, β b, B hctis but not limited thereto. The lactic acid bacteria used for fermentation in the method indicated by the present invention may use a single lactic acid strain, or a mixture The above two methods are often applied to the preparation of φ of the conventional fermented milk, and thus are not particularly limited in the present invention. The enzyme t in the present invention is not produced here, and any enzyme known or unknown that can be applied to hydrolyzed proteins can be used in the present invention, and examples exemplified herein include Prozyme 6 ®, Flav bribe yme®, Pr〇t_妒, etc., but not limited thereto. The concentration of the aforementioned enzyme is preferably 0.1 to 1.0% relative to the total amount of protein in the matrix), more preferably G.3 to 〇.7 % (w/w). Those skilled in the art of the present invention can also easily obtain the oligopeptides of the present invention by artificial synthesis by conventional peptide synthesis methods, or by (4) knowing the New Synthesizer, by reading the present specification. . ... The conversion of the hair to the vascular age is a competitive suppression $ state. After inhibition by in vitro enzymes, it was confirmed that the present invention can inhibit the action of vasopressin-converting enzyme. Example 1, +, dressing buffer, gluten A (l mM boric acid buffer solution, ρ Η 8 · 3) and buffered liquid B (3 600 m] y [1 mM chlorinated buffer solution of sodium chloride , ρΗ8 3). '1297272 Prepare jk tube vasopressin converting enzyme (ACE) solution · Add 5 units of vasopressin-converting enzyme powder to 5 〇ml of buffer solution A, and add equal amount of buffer solution B when used. To obtain a 5 〇 mU/ml vasopressin-converting enzyme solution. Hippuric acid-histamine-His-Leu (HHL) / gluten solution I · 500mg HHL powder, each added 38.8ml buffer solution A and B, mixed evenly to obtain 15mM HHL Solution. In addition, the oligopeptide was invented by the peptide synthesizer. At the time of the measurement, the oligopeptide of the present invention was dissolved in 5 ml of the buffer solution A and 5 ml of the buffer solution B. Thereafter, 75 melons were taken out therefrom, mixed with 75 μί of the acE solution, and preheated at 37 ° C for 1 minute, and then 15 mM of HHL solution was added as a substrate. After the mixture was reacted in a 3 rt water bath for 3 minutes, the reaction was quenched by the addition of 0.25 ml of 1N hydrochloric acid. The resulting hippuric acid was detected by high performance liquid chromatography (HPLC) at 228 nm to determine the percentage of inhibition of ACE activity. The calculation formula is as follows: Inhibition ability (%) = [(Ac_As) / (Ac_Ab)] χΐ〇〇
Ac=以緩衝液取代本發明寡胜肽反應後之吸光值Ac=the absorbance after the oligopeptide reaction of the present invention is replaced by a buffer
As=以本發明寡胜肽進行反應後之吸光值 AB=以緩衝液取代本發明寡胜肽，反應前先加鹽酸中止 反應後所測得之吸光值 1(^5〇值的測定： 測定對A C E的抑制能力之後，將濃度取對數與抑制能 作圖，求得迴歸曲線，並依此回歸方程式，可求得抑制 CE活性5〇%所需之乳清濃度，即為IQ。值。IQ。值越低’ ,'1297272 表示抑制ACE的效果越強。 值為 90·9μΜ。As = absorbance after reaction with the oligopeptide of the present invention AB = substitution of the oligopeptide of the present invention with a buffer, and the absorbance measured after the reaction was stopped by adding hydrochloric acid before the reaction 1 (measurement of the value of 〇5 :: determination After the inhibition ability of ACE, the logarithm of the concentration and the inhibition energy can be plotted, and the regression curve is obtained, and according to the regression equation, the whey concentration required to inhibit the CE activity of 5〇% can be obtained, that is, the IQ value. IQ. The lower the value, '1297272, the stronger the effect of suppressing ACE. The value is 90·9μΜ.
本發明寡胜肽經計算其1CThe oligopeptide of the present invention is calculated by its 1C
10 ,1297272 【圖式簡單說明】 無 【主要元件符號說明】 無10 ,1297272 [Simple diagram description] None [Main component symbol description] None