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| 亚麻籽巯基肽的制备及富集 |
| Preparation and enrichment of flaxseed sulfhydryl peptide |
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| DOI： |
| 中文关键词: 亚麻籽巯基肽 制备 富集 共价色谱 |
| 英文关键词:flaxseed sulfhydryl peptide preparation enrichment covalent chromatography |
| 基金项目: |
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| 中文摘要: |
| 天然巯基化合物因生理活性高、毒副作用低等优势越来越受到关注。为获得天然巯基肽，以低温压榨亚麻籽饼为原料提取亚麻籽蛋白，进一步酶解制备富含巯基的亚麻籽蛋白酶解物，并采用共价色谱法富集亚麻籽巯基肽。首先对蛋白酶进行了筛选，然后以酶解温度、pH、酶解时间、酶添加量、料液比为考察因素，以巯基含量为评价指标，在单因素实验的基础上通过正交实验对酶解工艺进行优化。采用Thiopropyl Crystarose 6 Fast Flow凝胶对亚麻籽蛋白酶解物中巯基肽进行选择性吸附并洗脱收集，并对亚麻籽巯基肽的氨基酸组成进行了分析。结果表明：最佳酶解工艺条件为采用木瓜蛋白酶、料液比1∶60、pH 7.0、酶解温度55℃、酶添加量3%、酶解时间4 h，在此条件下亚麻籽蛋白酶解物的巯基含量为16.69 μmol/g；通过富集得到巯基含量为71.13 μmol/g的亚麻籽巯基肽；相较于亚麻籽蛋白，亚麻籽巯基肽中含巯基氨基酸比例升高。综上，以低温压榨亚麻籽饼为原料，通过蛋白提取、酶解和共价色谱法富集可得到亚麻籽巯基肽。 |
| 英文摘要: |
| Natural sulfhydryl compounds have attracted more and more attention because of their advantages of high physiological activity and low toxicity and side effects. In order to obtain natural sulfhydryl peptides, flaxseed protein was extracted from low-temperature pressed flaxseed cake, and further enzymatically hydrolyzed to prepare flaxseed protein hydrolysates rich in sulfhydryl. Flaxseed sulfhydryl peptide was enriched by covalent chromatography. Firstly, the protease was screened, and then the enzymatic hydrolysis process was optimized by single factor experiment and orthogonal experiment with enzymatic hydrolysis temperature, pH, enzymatic hydrolysis time, enzyme dosage and solid-liquid ratio as investigation factors and sulfhydryl content as evaluation index. Thiopropyl Crystarose 6 Fast Flow gel was used for selective adsorption and desorption of sulfhydryl peptides from flaxseed protein hydrolysates, and the amino acid composition of flaxseed sulfhydryl peptide was analyzed. The results showed that the optimal enzymatic hydrolysis conditions were obtained as follows: using papain, enzymatic hydrolysis temperature 55℃, pH 7.0, enzymatic hydrolysis time 4 h, enzyme dosage 3% and solid-liquid ratio 1∶60. Under these conditions, the sulfhydryl content in flaxseed protein hydrolysates was 16.69 μmol/g, and it was 71.13 μmol/g after enrichment. Compared with flaxseed protein, the proportion of sulfhydryl amino acids in flaxseed sulfhydryl peptides increased. In summary, with low-temperature pressed flaxseed cake as raw material, flaxseed sulfhydryl peptide can be obtained by protein extraction, enzymatic hydrolysis and covalent chromatography enrichment. |
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