{"title":"大肠杆菌中黄嘌呤氧化酶抑制肽的序列优化和异源表达","authors":"","doi":"10.1016/j.fbio.2024.105080","DOIUrl":null,"url":null,"abstract":"<div><p>To solve the problem of low efficiency and purity in preparation of active peptides through enzymatic hydrolysis, the xanthine oxidase (XO) inhibitory peptides were optimized according to structure-activity relationship and a heterologous expression system for these peptides was constructed. The XO inhibitory peptide AEAWMWR (IC<sub>50</sub> = 1.76 mM), which exhibited enhanced activity, was obtained by optimizing AEAQMWR (IC<sub>50</sub> = 8.85 mM) in this research. The optimized peptide AEAWMWR exhibited approximately a 5-fold increase in activity compared to the template peptide AEAQMWR. The optimization results indicated that replacing the non-hydrophobic amino acids in the middle of the sequence with W or adding W to the C-terminal of the sequence effectively improved the activity of peptides. Additionally, to further achieve low-cost and rapid preparation of the peptides AEAQMWR and AEAWMWR, the recombinant plasmids containing fusion proteins of tandem repetitive peptides were designed and expressed in <em>Escherichia coli</em>. The recombinant peptides AEAQMWR (IC<sub>50</sub> = 8.19 mM) and AEAWMWR (IC<sub>50</sub> = 1.57 mM) exhibited significant activity. These results demonstrate that rational optimization and microbial synthesis of peptides can efficiently prepare bio-active peptides.</p></div>","PeriodicalId":12409,"journal":{"name":"Food Bioscience","volume":null,"pages":null},"PeriodicalIF":4.8000,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2212429224015104/pdfft?md5=a31be6a425c6b238d03b83410ada846a&pid=1-s2.0-S2212429224015104-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Sequence optimization and heterologous expression of xanthine oxidase inhibitory peptides in Escherichia coli\",\"authors\":\"\",\"doi\":\"10.1016/j.fbio.2024.105080\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>To solve the problem of low efficiency and purity in preparation of active peptides through enzymatic hydrolysis, the xanthine oxidase (XO) inhibitory peptides were optimized according to structure-activity relationship and a heterologous expression system for these peptides was constructed. The XO inhibitory peptide AEAWMWR (IC<sub>50</sub> = 1.76 mM), which exhibited enhanced activity, was obtained by optimizing AEAQMWR (IC<sub>50</sub> = 8.85 mM) in this research. The optimized peptide AEAWMWR exhibited approximately a 5-fold increase in activity compared to the template peptide AEAQMWR. The optimization results indicated that replacing the non-hydrophobic amino acids in the middle of the sequence with W or adding W to the C-terminal of the sequence effectively improved the activity of peptides. Additionally, to further achieve low-cost and rapid preparation of the peptides AEAQMWR and AEAWMWR, the recombinant plasmids containing fusion proteins of tandem repetitive peptides were designed and expressed in <em>Escherichia coli</em>. The recombinant peptides AEAQMWR (IC<sub>50</sub> = 8.19 mM) and AEAWMWR (IC<sub>50</sub> = 1.57 mM) exhibited significant activity. These results demonstrate that rational optimization and microbial synthesis of peptides can efficiently prepare bio-active peptides.</p></div>\",\"PeriodicalId\":12409,\"journal\":{\"name\":\"Food Bioscience\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.8000,\"publicationDate\":\"2024-09-08\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2212429224015104/pdfft?md5=a31be6a425c6b238d03b83410ada846a&pid=1-s2.0-S2212429224015104-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Food Bioscience\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2212429224015104\",\"RegionNum\":1,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"FOOD SCIENCE & TECHNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Food Bioscience","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2212429224015104","RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"FOOD SCIENCE & TECHNOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
为解决酶水解制备活性肽效率低、纯度低的问题,根据结构-活性关系对黄嘌呤氧化酶(XO)抑制肽进行了优化,并构建了这些肽的异源表达系统。在这项研究中,通过优化 AEAQMWR(IC50 = 8.85 mM),获得了具有更高活性的 XO 抑制肽 AEAWMWR(IC50 = 1.76 mM)。与模板肽 AEAQMWR 相比,优化后的肽 AEAWMWR 的活性提高了约 5 倍。优化结果表明,将序列中间的非疏水氨基酸替换为 W 或在序列的 C 端添加 W 能有效提高多肽的活性。此外,为了进一步实现低成本、快速制备多肽 AEAQMWR 和 AEAWMWR,设计了含有串联重复多肽融合蛋白的重组质粒,并在大肠杆菌中表达。重组肽 AEAQMWR(IC50 = 8.19 mM)和 AEAWMWR(IC50 = 1.57 mM)表现出显著的活性。这些结果表明,对多肽进行合理优化和微生物合成可有效制备具有生物活性的多肽。
Sequence optimization and heterologous expression of xanthine oxidase inhibitory peptides in Escherichia coli
To solve the problem of low efficiency and purity in preparation of active peptides through enzymatic hydrolysis, the xanthine oxidase (XO) inhibitory peptides were optimized according to structure-activity relationship and a heterologous expression system for these peptides was constructed. The XO inhibitory peptide AEAWMWR (IC50 = 1.76 mM), which exhibited enhanced activity, was obtained by optimizing AEAQMWR (IC50 = 8.85 mM) in this research. The optimized peptide AEAWMWR exhibited approximately a 5-fold increase in activity compared to the template peptide AEAQMWR. The optimization results indicated that replacing the non-hydrophobic amino acids in the middle of the sequence with W or adding W to the C-terminal of the sequence effectively improved the activity of peptides. Additionally, to further achieve low-cost and rapid preparation of the peptides AEAQMWR and AEAWMWR, the recombinant plasmids containing fusion proteins of tandem repetitive peptides were designed and expressed in Escherichia coli. The recombinant peptides AEAQMWR (IC50 = 8.19 mM) and AEAWMWR (IC50 = 1.57 mM) exhibited significant activity. These results demonstrate that rational optimization and microbial synthesis of peptides can efficiently prepare bio-active peptides.
Food BioscienceBiochemistry, Genetics and Molecular Biology-Biochemistry
CiteScore
6.40
自引率
5.80%
发文量
671
审稿时长
27 days
期刊介绍:
Food Bioscience is a peer-reviewed journal that aims to provide a forum for recent developments in the field of bio-related food research. The journal focuses on both fundamental and applied research worldwide, with special attention to ethnic and cultural aspects of food bioresearch.