脱脂豆粕酶法生产大豆肽工艺条件的研究

采用酶法水解脱脂豆粕制备大豆肽,确定了蛋白的浸出温度,比较单双酶的水解效果.确定酶解最佳条件：中性酶、复合酶添加量分别为400,200 U（每克蛋白中）,酶解温度为55℃,pH值为7.5,时间为2h,大豆肽产率约为32%.     

抗氧化作用延缓疲劳的研究进展

对抗氧化作用的机理进行了分析,抗氧化酶和非酶类物质是延缓疲劳的主要物质。抗氧化物质之间的协同作用可以增强抗疲劳作用,抗氧化物质对慢性疲劳综合症具有治疗作用。探讨了测定氧化应激的方法,包括直接测定自由基的含量,由自由基引起的生物大分子的氧化损害,抗氧化物质的活性等,一种方法不能有效评价抗氧化水平,通过多个指标的测定可以得到较好的结果。此外,对抗氧化物质在延缓疲劳中的应用进行了论述。大豆肽、乳清肽、鱼蛋白肽和猪脾脏蛋白肽等抗氧化活性肽都是良好的缓解疲劳的物质。植物提取物如皂甙、活性多糖、类黄酮、多酚、茶碱及其他动植物和食用菌等物质也有应用于延缓疲劳的报道。     

Effect of sonication on thermolysin hydrolysis of ovotransferrin

The effect of sonication on proteolytic hydrolysis of ovotransferrin and bioactivities of the hydrolysates were investigated, and the large peptide fragments left in the hydrolysate were characterised. The results showed that sonication could increase the reactive sulphydryl groups in 5% ovotransferrin solution by 50%, although there were no improvements in the overall degree of hydrolysis. Furthermore, SDS–PAGE and reverse-phase HPLC profiles did not show difference of the treated samples. Angiotensin-converting enzyme (ACE) inhibitory activity, but not antioxidant activity, of the thermolysin hydrolysate was improved in a dose dependent manner at prolonged sonication time. Matrix assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF/MS) analysis revealed that thermolysin hydrolysate was composed of 926 peptides, of 99% having masses lower than 10 kDa and of 1% larger than 10 kDa derived from both N- and C-lobes of ovotransferrin.     

Hypotensive effect of a sweetpotato protein digest in spontaneously hypertensive rats and purification of angiotensin I-converting enzyme inhibitory peptides

We have investigated angiotensin I-converting enzyme (ACE) inhibitory activity in an enzyme digest of sweetpotato protein, the antihypertensive effect of the digest in spontaneously hypertensive rats (SHR), and the identification of an ACE inhibitory peptide. Protein was prepared from squeezed juice of sweetpotato by isoelectric focusing precipitation. Three kinds of proteases were selected for effective protein digestion. The digest, sweetpotato peptide (SPP), exhibited strong ACE inhibitory activity (IC₅₀: 18.2 μg/ml). SPP was orally administered by gavage to SHR at a dose of 100 mg/kg or 500 mg/kg. The systolic blood pressure and the diastolic blood pressure were measured at 0 (before administration), 2, 4, 8, and 24 h after administration. A dose-dependent decrease in systolic blood pressure in SHR was observed after oral administration of SPP. Significant differences between SPP-administered rats and control rats were observed 4 and 8 h after administration in the 500 mg/kg-administered group and 8 h after administration in the 100 mg/kg-administered group. Diastolic blood pressure also decreased in the SPP-administered groups, although the difference between SPP-administered rats and control rats was not significant. These results suggest that SPP may be useful in the prevention or treatment of hypertension. Peptides with ACE inhibitory activity were purified from SPP by absorption chromatography and preparative HPLC using an ODS column. The amino acid sequences of isolated peptides were I-T-P, I-I-P, G-Q-Y and S-T-Y-Q-T; their ACE inhibitory activities (IC₅₀) were 9.5 μM, 80.8 μM, 52.3 μM and 300.4 μM, respectively. In conclusion, I-T-P is a novel, strong ACE inhibitory peptide.     

拟穴青蟹原肌球蛋白抗原表位适配体小肽的筛选与鉴定

目的筛选对拟穴青蟹过敏原原肌球蛋白(tropomyosin,TM)抗原表位具有特异性结合能力的适配体小肽,并鉴定适配体小肽对TM的免疫结合活性的抑制作用。方法采用ExPASy peptide cutter软件预测TM分子中胰蛋白酶酶切位点,结合TM线性表位设计反义小肽,经AutoDock 4.0软件模拟分子对接,筛选得到适配体小肽。采用抑制性ELISA的方法,检测筛选的适配体小肽对TM与特异性IgG抗体结合活性的抑制作用。结果 TM氨基酸序列中有25个胰蛋白酶酶切位点,设计TM的31条反义小肽,利用分子对接筛选得到12个适配体小肽。抑制性ELISA结果显示,12条适配体小肽均能明显抑制TM与IgG抗体的特异性结合,其中适配体小肽5抑制能力最强,其抑制率为36.2%。结论通过分子对接筛选得到能明显抑制TM免疫结合活性的适配体小肽,为降低TM致敏性的研究提供理论参考。     

酶法制备花生粕醒酒多肽

为更合理有效地利用花生粕资源,制备具有醒酒作用的花生粕多肽,本实验以Alcalase AF 2.4L蛋白酶酶法制备花生粕醒酒肽,经分级分离后,通过体外和动物实验验证其醒酒效果。结果表明,花生粕醒酒肽制备最佳工艺条件为：酶解时间3 h、酶解温度35 ℃、pH 9.5及料液比1∶30（m/V）。该条件下乙醇脱氢酶（alcoholdehydrogenase,ADH）激活率及多肽得率均较高。经分级分离后,分子质量在1 000～3 000 D的多肽对ADH激活作用最强,激活率为30.47%。G25凝胶色谱分析表明,花生粕多肽中小于3 000 D的多肽占89.55%,ADH激活率为29.25%。动物实验证明,花生粕多肽对小鼠有显著的防醉醒酒作用。小鼠血液乙醇含量测定结果表明,高剂量的花生粕多肽在60～90 min内能显著降低小鼠血液中的乙醇含量。     

酶法制备小分子大豆肽的蛋白酶筛选以及酶解条件优化

本研究对制备小分子大豆肽的蛋白酶进行筛选,并对酶解条件进行优化。综合考虑酶量对产物的水解度、平均分子量及游离氨基酸的影响,确定酶解的最佳条件为:温度55℃,pH值8.5,底物浓度为10%,分别加入1.5%的木瓜蛋白酶、菠萝蛋白酶、无花果蛋白酶和3942中性蛋白酶,调pH值至4.5,再分别加入2.5%的Fromase真菌蛋白酶和537酸性蛋白酶。结果显示,多酶复合水解可提高蛋白质降解率,并尽可能降低多肽的相对分子质量。     

罗非鱼皮胶原肽的制备及抗氧化活性研究

采用酶法从罗非鱼皮中制取胶原肽,选用动物蛋白水解酶,当酶的添加量为0.5%,61℃,pH 6.7下水解3 h时,蛋白质的回收率为92.97%,水解度为9.37%.水解产物90%以上相对分子质量低于10 kD的胶原肽.胶原肽对由Fe^2＋引发的卵磷脂脂质体过氧化有一定抑制作用,同时对·OH具备一定的清除作用.     

大枣多肽裂解液抗肿瘤作用研究

目的:探讨大枣多肽裂解液抗肿瘤作用并初步探讨其抗肿瘤机制,为其临床应用提供实验室依据。方法:将接种肿瘤后的小鼠随机分为5组,每组10只小鼠,雌雄各半,给药10d,停药后24h处死动物,称小鼠体重,剥离小鼠右腋下皮下组织,摘出肿瘤,称瘤重,测定生化指标。结果:对肉瘤(S180)、肝癌(Hep)、胃癌(MFC)细胞荷瘤小鼠肿瘤生长具有明显的抑制作用,其中对S180作用显著,较低剂量即可显示明显作用,延长S180荷瘤鼠生存时间;同时可增加吞噬指数α、廓清指数K、及胸腺系数;对体外淋巴细胞转化功能亦有增强作用。结论:大枣多肽裂解液具有明显抗肿瘤活性。