| 祖丽比娜·艾尼瓦尔，徐同翔，罗春燕，许世恒，王彩蝶.中性蛋白酶酶解棉籽蛋白条件优化及其代谢物的变化[J].中国油脂,2026,51(1):.[ZULIBINA Ainiwaer，XU Tongxiang，LUO Chunyan，XU Shiheng，WANG Caidie.Optimization of enzymatic hydrolysis conditions of cottonseed protein by neutral protease and changes of its metabolites[J].China Oils and Fats,2026,51(1):.] |
| 中性蛋白酶酶解棉籽蛋白条件优化及其代谢物的变化 |
| Optimization of enzymatic hydrolysis conditions of cottonseed protein by neutral protease and changes of its metabolites |
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| DOI：10.19902/j.cnki.zgyz.1003-7969.240651 |
| 中文关键词: 中性蛋白酶 棉籽蛋白 水解度 代谢物 非靶向代谢组学 |
| 英文关键词:neutral protease cottonseed protein degree of hydrolysis metabolites non-targeted metabolomics |
| 基金项目:自治区重点研发计划项目（2022B02057-1-3）；国家自然科学基金项目（32160811）；新疆农业大学自治区级大学生创新训练计划项目（S202410758010）；新疆农业大学博士后流动站资助（2021年） |
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| 中文摘要: |
| 旨在为棉籽蛋白在饲料领域中的应用提供理论依据与技术支撑，以棉籽蛋白为原料，使用中性蛋白酶对其进行酶解，采用单因素试验研究酶解时间、酶解温度、料液比、酶添加量对蛋白水解度的影响。在此基础上，采用正交试验进行工艺优化，检测最佳酶解条件下酸溶蛋白和小肽含量及游离氨基酸组成变化，并对代谢物进行非靶向代谢组学分析。结果表明：中性蛋白酶酶解棉籽蛋白的最佳条件为酶解温度50 ℃、酶解时间48 h、料液比1∶ 1、酶添加量1 500 U/g，在此条件下水解度为1405%，小肽、酸溶蛋白及游离氨基酸总含量较未添加蛋白酶空白组棉籽蛋白分别提高了33.51百分点、3632百分点、28.32 mg/g（p＜0.01）；非靶向代谢组学结果显示，共鉴定出14类代谢物、1 021个差异代谢物，主要富集在次生代谢物的生物合成和嘌呤代谢，且代谢物中包含具有抗炎、抗氧化等作用的活性物质。综上，利用中性蛋白酶酶解棉籽蛋白可以显著提高棉籽蛋白的水解度、酸溶蛋白以及游离氨基酸含量，且代谢物中含有多种生物活性物质，从而提高了棉籽蛋白的营养价值与功能特性。 |
| 英文摘要: |
| The aim is to provide theoretical foundations and technical support for the application of cottonseed protein in feed. Using cottonseed protein as raw material, the hydrolysis with neutral protease was performed. The effects of hydrolysis time, hydrolysis temperature, solid-liquid ratio and enzyme addition amount on degree of hydrolysis (DH) were studied by single factor experiments. Based on this, the hydrolysis process were optimized by orthogonal experiment, and the changes of acid-soluble protein and peptide contents, as well as free amino acids composition under the optimal conditions were detected. Non-targeted metabolomics analysis was performed on the metabolites. The results showed that the optimum parameters for enzymatic hydrolysis of cottonseed protein by neutral protease were as follows : hydrolysis temperature 50 ℃, hydrolysis time 48 h, solid-liquid ratio 1∶ 1, and enzyme addition amount 1 500 U/g. Under the optimal conditions, DH reached 14.05%, with peptide and acid-soluble protein contents, as well as total free amino acids, increasing by 33.51 percentage points, 36.32 percentage points and 28.32 mg/g, respectively, compared to cottonseed protein control without protease(p<001). Non-targeted metabolomics results revealed 14 metabolite classes and 1 021 differentially expressed metabolites, primarily enriched in biosynthesis of secondary metabolites and purine metabolism. These metabolites included bioactive compounds with anti-inflammatory and antioxidant properties. In summary, the enzymatic hydrolysis of cottonseed protein using neutral protease can significantly increase the DH, acid-soluble protein content, and free amino acid content of cottonseed protein. Moreover, the metabolites contain multiple bioactive compounds, thereby improving the nutritional value and functional properties of cottonseed protein. |
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