本文選題：薏芯仁 + 酶法��； 參考：《黑龍江中醫(yī)藥大學(xué)》2010年碩士論文

【摘要】：本論文采用生物轉(zhuǎn)化方法制備薏苡仁抗高血壓肽,初步確定了薏苡仁活性肽的體外抗高血壓活性,為抗高血壓藥的進(jìn)一步研究提供參考。同時比較了酶法與發(fā)酵法制備薏苡仁肽的體外抗高血壓活性,結(jié)果證明發(fā)酵法得到的活性肽具有更強(qiáng)的活性。 本實(shí)驗分別進(jìn)行了酶解法和發(fā)酵制備薏苡仁肽兩方面的研究。優(yōu)化了薏苡仁蛋白質(zhì)水提和酶解條件,結(jié)果表明,蛋白質(zhì)水提最佳條件為pH=10,溫度70℃,料水比1：15；堿性蛋白酶(Alcalase)對薏苡仁的酶解效果最好,酶解最佳條件為加酶量5%,時間2h,料水比1：25,此時的多肽含量遠(yuǎn)遠(yuǎn)高出水解時的含量。用樹脂對酶解液進(jìn)行純化,大孔吸附樹脂DA201-C對酶解液的吸附效果較好；當(dāng)酶解產(chǎn)物pH為5.5,上樣流速1BV/h時,吸附率最高；當(dāng)用75%乙醇進(jìn)行解吸時,解吸時間和濃縮時間短,解吸率高。使用高效液相色譜(HPLC)測定樹脂純化后產(chǎn)物的分子量,分子量在278.61至943.32之間的物質(zhì)占純化后產(chǎn)物的99.3%。用葡聚糖凝膠Sephadex G-15對純化產(chǎn)物進(jìn)行分離,以去離子水為緩沖液時可以得到三個峰,其中B峰具有最高的ACE抑制率,為80.1%。 優(yōu)化了薏苡仁蛋白質(zhì)發(fā)酵條件,結(jié)果表明,三種曲霉分別發(fā)酵中藥薏苡仁,活性肽含量黑曲霉達(dá)到6.14mg/ml;米曲霉達(dá)到2.58mg/ml;紅曲霉分別為2.93mg/ml,因此確定黑曲為最佳菌種進(jìn)行混合發(fā)酵。最佳條件為發(fā)酵時間90h,薏苡仁料液濃度9%,PH=5,接種量11%�；旌习l(fā)酵對黃酒酵母、酒精酵母、啤酒酵母進(jìn)行篩選,確定黃酒酵母為最佳菌種。用樹脂對酶解液進(jìn)行純化。使用HPLC測定樹脂純化后產(chǎn)物的分子量在267.91至930.35之間的物質(zhì)占純化后產(chǎn)物的98.4%。用葡聚糖凝膠Sephadex G-15對純化產(chǎn)物進(jìn)行分離,以去離子水為緩沖液時可以得到五個峰,其中B峰具有最高的ACE抑制率,為83.2%,體外抗高血壓活性與酶解薏苡仁活性肽相比有所提高,因此發(fā)酵法比酶解效果更好。
[Abstract]:In this paper, the antihypertensive peptide of Coix seed was prepared by biotransformation, and the in vitro antihypertensive activity of Coix seed active peptide was preliminarily determined, which provides a reference for the further study of antihypertensive drugs. At the same time, the antihypertensive activity of Coix seed peptide prepared by enzymatic method and fermentation method was compared in vitro. The results showed that the active peptide obtained by fermentation method had stronger activity. The enzymatic hydrolysis and fermentation of Coix seed peptide were studied in this experiment. The optimum conditions for protein extraction and enzymatic hydrolysis of Coix seed were optimized. The results showed that the optimum conditions of protein extraction were pH 10, temperature 70 鈩，

本文編號：1781520