| 摘要: |
| 目的：优化黄精多肽的制备工艺，并对所得黄精多肽进行鉴定与活性筛选，以期获得高活性的黄精多肽。方法：通过单因素与响应面实验优化黄精多肽的制备工艺；采用LC-MS/MS技术鉴定黄精多肽，并利用PeptideRanker、ToxinPred和AntiCP进行活性筛选。结果：黄精多肽制备方法为酶-菌协同法，最佳工艺条件为碱性蛋白酶添加量0.75%、起始pH 5.0、植物乳杆菌添加量1.50%、发酵温度35 ℃，在此条件下进行3次平行试验，发酵12 h后黄精多肽得率为（32.12 ± 0.40） mg/g。酶-菌协同法共鉴定出133个黄精多肽，而酶解法则仅鉴定出67个，前者肽段数量增加66个，且多肽丰度为酶解法的1.25倍。此外，酶-菌协同法获得14个高活性肽段，酶解法仅获得7个。结论：酶-菌协同法有助于黄精多肽的制备，可显著提高肽段的数量和丰度，促进高活性肽的产生。本研究优化了黄精多肽的制备工艺，其活性预测结果可为黄精多肽功能产品的开发提供参考。 |
| 关键词: 黄精 生物活性肽 酶-菌协同法 工艺优化 活性筛选 响应面 |
| DOI： |
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| Optimization of the preparation process of bioactive peptides from Polygonatum sibiricum and an activity screening study |
| LIU Yujun,LI Yanping,HE Wei,JIN Jian,LAO Jia,PAN Gen,ZHANG Shuihan,ZHONG Can |
| (Institute of Chinese Medicinal Resources, Hunan Academy of Chinese Medicine, Changsha 410013, Hunan, China;Institute of Chinese Medicinal Resources, Hunan Academy of Chinese Medicine, Changsha 410013, Hunan, China; Hunan University of Chinese Medicine, Changsha 410208, Hunan, China;RESGREEN Biotechnology Group Co., Ltd., Changsha 410329, Hunan, China) |
| Abstract: |
| Objective: To optimize the preparation process of Polygonatum sibiricum polypeptides, to perform identification and activity screening of the Polygonatum sibiricum polypeptides obtained, and to acquire highly active Polygonatum sibiricum polypeptides. Methods: Single-factor experiments and response surface methodology were used to optimize the preparation process of Polygonatum sibiricum polypeptides. LC-MS/MS was used for identification of Polygonatum sibiricum polypeptides, and PeptideRanker, ToxinPred, and AntiCP were used for activity screening. Results: The enzyme-microbe synergistic method was used to prepare Polygonatum sibiricum polypeptides, and the optimal processing conditions were determined as an additive amount of 0.75% for alkaline protease, an initial pH value of 5.0, an additive amount of 1.50% for Lactobacillus plantarum, and a fermentation temperature of 35℃. Three parallel experiments were conducted under these conditions, and the yield of Polygonatum sibiricum polypeptides was （32.12±0.40 mg/g） after 12 hours of fermentation. A total of 133 Polygonatum sibiricum polypeptides were identified by the enzyme-microbe synergistic method, whereas only 67 were identified by the enzymatic hydrolysis method, suggesting that the number of peptides was increased by 66 based on the former method, and the peptide abundance of the enzyme-microbe synergistic method was 1.25 times that of the enzymatic hydrolysis method. In addition, the enzyme-microbe synergistic method yielded 14 highly active peptides, while the enzymatic hydrolysis method yielded only 7 highly active peptides. Conclusion: The enzyme-microbe synergistic method can facilitate the preparation of Polygonatum sibiricum polypeptides by significantly improving the quantity and abundance of peptides and promoting the production of highly active peptides. This study optimized the preparation process of Polygonatum sibiricum polypeptides, and the results of activity prediction can provide a reference for the development of functional products derived from Polygonatum sibiricum polypeptides. |
| Key words: Polygonatum sibiricum bioactive peptides enzyme-microbe synergistic method process optimization activity screening response surface methodology |
