| 摘要: |
| 为探究胰蛋白酶、胃蛋白酶和木瓜蛋白酶酶解奶牛胎盘产物间差异，用此3种酶在最优条件下酶解奶牛胎盘获得胎盘酶解产物，每种酶重复3次，利用LC-MS/M技术鉴定产物中蛋白和肽段，BIO-PEP数据库匹配和测算十肽及以下的抗氧化活性多肽。结果显示:胰蛋白酶、胃蛋白酶和木瓜蛋白酶酶解产物分别鉴定出541、137和86种蛋白，2 766、1 170和128个肽段，胰蛋白酶鉴定蛋白和肽段数极显著高于其余2种酶(P<0.01)，木瓜蛋白酶鉴定蛋白和肽段数均显著低于其余2种酶(P<0.05)；3种酶酶解产物十肽及以下具有潜在抗氧化能力肽段分别占十肽及以下肽段总量的39.14%(256/654)、32.80%(81/247)和53.33%(16/30)，木瓜蛋白酶占比显著高于其余2种酶(P<0.05)；胰蛋白酶、胃蛋白酶和木瓜蛋白酶3种酶酶解产物抗氧化多肽共同特征是分别含有EL、LY、MM、LK、IR、MY、VY残基，HL、LK、TY、HH、EL残基和LK、IR、EL等残基。综上，通过LC-MS/M技术对酶解产物定性和定量分析，胰蛋白酶酶解奶牛胎盘的产物生物学信息最丰富，在蛋白组学中可靠性和准确性最高；木瓜蛋白酶酶解奶牛胎盘最彻底，其产物潜在抗氧化活性最强，本研究为使用蛋白组学研究奶牛胎盘时酶的选择和奶牛胎盘抗氧化活性多肽的开发利用提供理论依据。 |
| 关键词: 奶牛 胎盘 蛋白酶 LC-MS/MS 抗氧化 |
| DOI：10.11841/j.issn.1007-4333.2021.11.13 |
| 投稿时间：2021-02-20 |
| 基金项目:四川省科技厅科技计划项目(2019YJ0650)；国家重点研发计划项目(2018YFD0500900)；江苏农牧科技职业学院产业发展关键技术创新项目(NSF2021ZR01)；四川农业大学本科科研兴趣培养计划(KY2020186) |
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| LC-MS/MS based comparative analysis and antioxidant activity prediction of cow placenta hydrolyzed by different proteases |
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ZHANG Yue1,YOU Liuchao1,ZHU Yinkun1,SHEN Yu1,LV Shangkui1,PEI Miaoyu1,FU Hongqing2,SHEN Liuhong1*
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| (1.The Key Laboratory of Animal Disease and Human Health of Sichuan Province/The Medical research center for cow disease/College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;2.Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China) |
| Abstract: |
| In order to investigate the differences among the placenta products of trypsin, pepsin and papain, placenta hydrolysates were obtained by enzyme hydrolysis of the three enzymes under the optimal conditions. Each enzyme was repeated 3 times. LC-MS/M technology was used to identify the proteins and peptides in the products, and the BIO-PEP database was used to match and calculate the antioxidant peptides below peptides. The results showed 541, 137, 86 proteins and 2 766, 1 170, 128 peptides were identified in trypsin group, pepsin group and papain group, respectively. The number of identified proteins and peptides in the trypsin group was significantly higher than the other two groups(P<0. 01), and the number of identified proteins and peptides in the papain group was significantly lower than the other two groups(P<0. 05). The potential antioxidant peptides of decapeptide and below accounted for 39. 14%(256/654), 32. 80%(81/247)and 53. 33%(16/30). The proportion of papain group was significantly higher than the other two groups(P<0. 05). The common features of three kinds of antioxidant peptides were EL, LY, MM, LK, IR, MY, VY, HL, LK, TY, HH, El and LK, IR, EL. In summary, the biological information of cow placenta hydrolyzed by trypsin is the richest, and its reliability and accuracy are the highest in proteomics. Papain has the most thorough enzymatic hydrolysis of cow placenta, and its products have the strongest potential antioxidant activity. The antioxidant activity of dairy cow placenta comes from Y, M, H, K. In summary, according to the qualitative and quantitative analysis of enzymolysis products by LC-MS/M technology, the biological information of cow placenta hydrolyzed by trypsin is the most abundant, and its reliability and accuracy are the highest in proteomics. Papain hydrolyzes cow placenta most thoroughly, and its products have the strongest potential antioxidant activity. This study provides a theoretical basis for the selection of enzymes and the development and utilization of antioxidant peptides in cow placenta by proteomics. |
| Key words: dairy cow placenta protease LC-MS/MS antioxidation. |
