【摘要】：1．乳源免疫調(diào)節(jié)肽的制備、純化及鑒定 目的 探討酪蛋白酶解產(chǎn)生免疫調(diào)節(jié)肽的最適條件，建立酶解產(chǎn)物的分離及其鑒定免疫調(diào)節(jié)肽的方法。方法 將酪蛋白酶解，其酶解產(chǎn)物經(jīng)超濾膜過濾，用十二烷基磺酸鈉——聚丙烯酰銨凝膠電泳(SDS-PAGE)方法分析其酶解情況。用反相高效液相色譜層析(RP-HPLC)，C_(18)柱，以免疫調(diào)節(jié)肽為標(biāo)準(zhǔn)已知樣，利用保留時(shí)間外標(biāo)法與內(nèi)標(biāo)法，與已知對照物進(jìn)行定性，鑒定酪蛋白酶解產(chǎn)物中的免疫調(diào)節(jié)肽。酶解上清液經(jīng)Sephadex G-15凝膠層析分離，得到不同級分，然后用RP-HPLC鑒定各級分中的免疫調(diào)節(jié)肽。結(jié)果 酪蛋白在胰蛋白酶與糜蛋白酶組合下水解90分鐘，能產(chǎn)生PGPIPN量最多，大約5％。酶解產(chǎn)物經(jīng)Sephadex G-15凝膠層析后，酶解產(chǎn)物中小分子物質(zhì)被分開，經(jīng)RP-HPLC檢測，Sephadex G-15的第5級分含有免疫調(diào)節(jié)肽。結(jié)論 酪蛋白經(jīng)過胰蛋白酶和糜蛋白酶酶解后可以生成免疫調(diào)節(jié)肽，并可以通過Sephadex G-15粗分免疫調(diào)節(jié)肽。 2．乳源免疫調(diào)節(jié)肽對大鼠生長和機(jī)體免疫的影響 目的 探討乳源免疫調(diào)節(jié)肽對大鼠機(jī)體免疫功能，增重和采食量的影響。方法 42只40日齡的SD大鼠(雌雄各一半)隨機(jī)分為兩個(gè)試驗(yàn)組和一個(gè)對照組，每組14只雌雄各一半，，分開飼養(yǎng)。預(yù)飼一周后，實(shí)驗(yàn)組分別被灌喂2.5×10~(-3)g·L~(-1)免疫調(diào)節(jié)肽(試驗(yàn)組Ⅰ)、2.5×10~(-2)g·L~(-1)免疫調(diào)節(jié)肽(試驗(yàn)組Ⅱ)：對照組灌喂生理鹽水，并記錄大鼠日采食量和體重。飼養(yǎng)一個(gè)月后分別檢測對照組和試驗(yàn)組的淋巴細(xì)胞轉(zhuǎn)化、紅細(xì)胞免疫、巨噬細(xì)胞吞噬、日采食量和體重等的差別。結(jié)果 乳源免疫調(diào)節(jié)肽對大鼠腹腔巨噬細(xì)胞吞噬和紅細(xì)胞免疫有顯著促進(jìn)功能(p＜0.05)；而對淋巴細(xì)胞轉(zhuǎn)化作用有增長的趨勢，但未達(dá)到顯著水平(p＞0.05)。與對照組相比，實(shí)驗(yàn)組Ⅱ的大鼠日采食量顯著地增加(P＜0.05)，雌雄大鼠分別提高了
[Abstract]:1. Preparation, purification and Identification of Milk-derived Immunoregulatory peptides objective to study the optimal conditions for casein enzymatic hydrolysis to produce immunomodulatory peptides. To establish a method for the isolation and identification of immunomodulatory peptides. Methods casein was hydrolyzed by ultrafiltration membrane, and the enzymatic hydrolysis of casein was analyzed by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunoregulatory peptides in casein enzymatic hydrolysis products were identified by reversed-phase high performance liquid chromatography (RP-HPLC) with C _ (18) column, using immunoregulatory peptides as standard known samples, using retention time external standard method and internal standard method, and qualitative analysis with known controls. The supernatant of enzymatic hydrolysis was separated by Sephadex G-15 gel chromatography, and the different fractions were obtained, and then the immunomodulatory peptides were identified by RP-HPLC. Results casein hydrolyzed in combination of trypsin and chymotrypsin for 90 minutes resulted in the highest amount of PGPIPN, about 5%. After Sephadex G-15 gel chromatography, the small molecules in the enzymatic hydrolysis products were separated. The fifth fraction of Sephadex G-15 contained immunomodulatory peptides by RP-HPLC. Conclusion casein can be hydrolyzed by trypsin and chymotrypsin to produce immunomodulatory peptide. The effect of milk source immunomodulatory peptide on the growth and body immunity of rats was studied. Objective to study the immune function of milk source immunomodulatory peptide in rats. The effects of weight gain and feed intake. Methods Forty-two 40-day-old SD rats (half of male and female) were randomly divided into two experimental groups and one control group. After one week of prefeeding, the experimental group was fed with 2.5 脳 10 ~ (-3) g L ~ (-1) immunomodulatory peptide (test group 鈪

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