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金枪鱼黑肉水解物抗氧化效果的分析与测试

analysis and tests of the tuna dark muscle hydrolysate anti-oxidation effect.

作者信息

Han Jiaojiao, Tang Shasha, Li Yanyan, Bao Wei, Wan Haitao, Lu Chenyang, Zhou Jun, Li Ye, Cheong Lingzhi, Su Xiurong

机构信息

School of Marine Science, Ningbo University 818 Fenghua Road Ningbo China

College of Agriculture and Life Sciences, Cornell University Ithaca USA.

出版信息

RSC Adv. 2018 Apr 17;8(25):14109-14119. doi: 10.1039/c8ra00889b. eCollection 2018 Apr 11.

Abstract

Hydrolysate is a mixture of various peptides with specific functions. However, functional identification of hydrolysate with high throughput is still a difficult task. Furthermore, using tests animal or cell experiments is time and labor-intensive. In this study, the peptides component of hydrolysate derived from the tuna dark muscle was measured MALDI-TOF/TOF-MS, and the functions of the KEFT (Lys-Glu-Phe-Thr), EEASA (Glu-Glu-Ala-Ser-Ala) and RYDD (Arg-Tyr-Asp-Asp) peptides, which were found with the highest proportion, were predicted Discovery Studio 2016 software. All three peptides were predicted to bind to the Keap1 protein with the highest fit-value and to affect the activity of Keap1, which is involved in anti-oxidation pathways. Subsequently, mice experiments showed that administration of tuna dark muscle hydrolysate increased the levels of superoxide dismutase and glutathione peroxidase in the serum and liver ( < 0.05) and decreased the malondialdehyde level ( < 0.05) as well as transcription of Keap1 ( > 0.05), which are consistent with the analysis results using Discovery Studio 2016 software. The combination of analysis and tests provided an alternative strategy for identifying hydrolysate function and provided insight into high-value utilization of protein hydrolysate.

摘要

水解产物是具有特定功能的各种肽的混合物。然而,高通量鉴定水解产物的功能仍然是一项艰巨的任务。此外,使用动物实验或细胞实验进行测试既耗时又费力。在本研究中,采用基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF/TOF-MS)测定了金枪鱼黑肉水解产物的肽成分,并使用Discovery Studio 2016软件预测了比例最高的KEFT(Lys-Glu-Phe-Thr)、EEASA(Glu-Glu-Ala-Ser-Ala)和RYDD(Arg-Tyr-Asp-Asp)肽的功能。预测这三种肽均以最高拟合值与Keap1蛋白结合,并影响参与抗氧化途径的Keap1的活性。随后,小鼠实验表明,给予金枪鱼黑肉水解产物可提高血清和肝脏中超氧化物歧化酶和谷胱甘肽过氧化物酶的水平(P<0.05),降低丙二醛水平(P<0.05)以及Keap1的转录水平(P>0.05),这与使用Discovery Studio 2016软件的分析结果一致。分析与实验相结合为鉴定水解产物功能提供了一种替代策略,并为蛋白质水解产物的高值利用提供了思路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3aa9/9079911/c44e6e9702aa/c8ra00889b-f1.jpg

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