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大豆肽的制備及其免疫調(diào)節(jié)作用的研究

發(fā)布時間:2018-10-05 09:19
【摘要】: 論文主要研究大豆肽的制備及其免疫調(diào)節(jié)功能,并初步探討大豆肽免疫活性與其電荷及相對分子質(zhì)量之間的關(guān)系。 通過單因素實驗制備了不同水解條件下水解的大豆肽,采用SP—Sephadex C-25陽離子交換樹脂測定水解肽正電荷肽含量,高效液相測定其相對分子質(zhì)量的分布,發(fā)現(xiàn)水解肽的相對分子質(zhì)量主要集中在1000以下,說明小肽是水解物中的主要成分。通過淋巴細胞增殖和巨噬細胞吞噬中性紅實驗,測得大豆肽對小鼠淋巴細胞增殖和巨噬細胞吞噬能力均有一定的促進作用,且大豆肽免疫活性大小與正電荷肽含量的相關(guān)性較好,而與平均分子量的相關(guān)性不大。其中,Alcalase為水解酶,酶用量為o.02g/g蛋白,底物濃度為5%,溫度60℃,pH值8.0,水解時間225min,活性最高。 通過給昆明種雄性小鼠(5.6周齡)連續(xù)3 d注射40mg/Kg的環(huán)磷酰胺,并灌胃16mg/Kg、40mg/Kg、80mg/Kg的大豆肽,發(fā)現(xiàn)大豆肽不僅能顯著提高正常小鼠免疫功能,而且能顯著提高環(huán)磷酰胺致免疫低下小鼠的臟器指數(shù),提高脾臟淋巴細胞轉(zhuǎn)化率、抗體生成細胞含量及血清溶血素含量(P0.05)。 用60mg/Kg的D-半乳糖連續(xù)42 d腹腔注射昆明種雄性小鼠(5.6周齡),同時灌胃16mg/Kg、40mg/Kg的大豆肽,結(jié)果表明,不同劑量大豆肽均能提高小鼠臟器指數(shù),脾臟淋巴細胞轉(zhuǎn)化率、抗體生成細胞含量及血清溶血素含量。
[Abstract]:In this paper, the preparation and immunomodulatory function of soybean peptide were studied, and the relationship between the immunological activity of soybean peptide and its charge and relative molecular weight was discussed. Soybean peptides hydrolyzed under different hydrolysis conditions were prepared by single factor experiment. The content of positively charged peptides was determined by SP-Sephadex C-25 cation exchange resin, and the distribution of relative molecular weight was measured by high performance liquid chromatography (HPLC). It was found that the molecular weight of hydrolyzed peptides was mainly below 1000, indicating that small peptides were the main components of hydrolysates. Through lymphocyte proliferation and macrophage phagocytosis neutral red test, it was found that soybean peptide could promote lymphocyte proliferation and macrophage phagocytosis in mice. The immunological activity of soybean peptide had a good correlation with the content of positively charged peptide, but had little correlation with the average molecular weight. Among them, Alcalase is hydrolase, enzyme dosage is o.02g/ g protein, substrate concentration is 5, temperature is 60 鈩,

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