Recent advances in synthesis and mining strategies of functional peptides

doi:10.12211/2096-8280.2024-067

Abstract

Abstract:

Functional peptide is a kind of short chain peptide composed of 2 to 50 amino acids, and its biological activity is closely related to the type, sequence, length and structure of the functional peptide itself. Functional peptides can play a regulatory role in a variety of physiological processes by specifically recognizing and binding to target molecules in vivo. Due to their rapid action, strong specificity and small toxic side effects, functional peptides have been widely concerned in the research and development of new drugs and functional raw materials, and have become a research hotspot in the field of life science and biotechnology. In this paper, functional peptides have shown great application potential in many fields such as biomedicine, food science and cosmetics. For example, in the field of biomedicine, functional peptides can be used as the basic material of antimicrobial, anticancer, immune regulation and other therapeutic factors. In the food industry, they are used as natural additives to enhance nutritional value and promote health. In the field of cosmetics, functional peptides are widely used for anti-aging, moisturizing and repairing the skin. Next, this paper discusses the ways of obtaining functional peptides, mainly including protein hydrolysis, chemical synthesis and biosynthesis (e.g., microbial recombinant expression technology), and compares the advantages and disadvantages of these two processes and their respective application scenarios. In terms of functional peptide mining strategies, this paper reviews the latest researches including phage surface display technology, machine learning algorithm, molecular docking technology and artificial intelligence technology. These techniques show significant potential in the screening and design of functional peptides, improving the efficiency and accuracy of research. In recent years, the rapid development of synthetic biology and the wide application of bioinformatics and artificial intelligence technologies have provided new ideas and strategies for the discovery and optimization of functional peptides, making it possible to screen functional peptides through machine learning and high throughput. Looking forward to the future, the research of functional peptides will face new challenges and opportunities. Improving the synthesis process to improve the efficiency, improving the stability of functional peptides through structural modification, and using computer-aided optimization and artificial intelligence to design multifunctional peptides will become important research directions. At the same time, strengthening the safety and efficacy assessment of functional peptides can further enhance the application potential of functional peptides.

Key words: functional peptide, synthetic biology, biosynthesis, high throughput, machine learning

摘要：

功能肽是由2至50个氨基酸组成的短链肽，近年来因其特异性强、作用迅速及副作用低而成为开发新药和功能原料的重要研究热点。首先，本文梳理了功能肽的分类、作用机制及应用场景，总结了不同类型功能肽的特点和在生物医药、食品科学及化妆品等领域的应用。接着，针对功能肽的合成方法，探讨了化学合成与生物合成的最新进展，比较了这两种制备工艺的优缺点以及各自的适用场景。在功能肽挖掘策略方面，本文综述了包括噬菌体表面展示技术、机器学习算法、分子对接技术及人工智能技术的最新研究，这些技术在功能肽的筛选和设计中展现出重要潜力，提升了研究的效率与准确性。展望未来，功能肽的研究将面临新的挑战与机遇。如何改进合成工艺以提高效率、如何通过结构修饰提高功能肽稳定性，以及如何利用计算机辅助优化和人工智能设计多功能肽，将成为重要的研究方向。同时，加强功能肽的安全性和有效性的评估能进一步提升功能肽的应用潜力。

关键词: 功能肽, 合成生物学, 生物合成, 高通量, 机器学习

CLC Number:

Q815

Chuangen TANG, Jing WANG, Shuo ZHANG, Haoning ZHANG, Zhen KANG. Recent advances in synthesis and mining strategies of functional peptides[J]. Synthetic Biology Journal, DOI: 10.12211/2096-8280.2024-067.

汤传根, 王璟, 张烁, 张昊宁, 康振. 功能肽合成和挖掘策略研究进展[J]. 合成生物学, DOI: 10.12211/2096-8280.2024-067.

Figures/Tables 6

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分类	作用机制和典型氨基酸序列	特点和应用场景	参考文献
抗菌肽	a.破坏细胞膜结构：通过与细菌细胞膜中的磷脂双层结合，增加膜通透性，导致细胞内容物泄漏，从而杀死细菌；b.抑制细菌代谢途径：干扰细菌DNA、RNA和蛋白质合成，抑制细菌生长和繁殖；典型氨基酸序列：Phe-Trp-Lys-Phe-Lys	广谱抗菌、抗感染，特别是对抗耐药性菌株感染有很好效果；伤口愈合；预防感染；女性阴道念珠菌感染等	[3-4]
抗病毒肽	a.阻断病毒进入细胞：通过与病毒外壳蛋白或宿主细胞受体结合，阻止病毒附着和进入宿主细胞；b.抑制病毒复制：干扰病毒RNA或DNA合成，从而抑制病毒复制；典型氨基酸序列：Ala-Ala-Val-Ala-Leu-Leu-Pro-Ala-Val--Leu-Leu-Ala-Leu-Leu-Ala-Pro	通过抑制病毒蛋白的功能，阻断病毒生命周期；可预防病毒感染，对于已感染病毒患者，则可阻止病毒在体内的进一步扩展	[4,19]
抗氧化肽	a.清除自由基：通过捐献电子或氢原子，中和体内自由基，减少自由基对细胞损伤；b.提高抗氧化酶活性：通过提高超氧化物歧化酶（Superoxide dismutase, SOD）和过氧化氢酶等抗氧化酶的活性，增强机体抗氧化能力；典型氨基酸序列：Leu-Ala-Asn-Ala-Lys	安全性高，可以用在食品领域抗氧化上；也可开发相应抗氧化化妆品、保健食品或者药品	[5-9]
抗紫外肽	a.吸收紫外线：通过吸收紫外线光子，减少其对皮肤的穿透和损伤；b.修复紫外线损伤：通过促进DNA修复机制，修复紫外线引起的DNA损伤，减少皮肤癌的发生；典型氨基酸序列：Leu-Val-Asn-Glu-Leu-Thr-Glu-Phe-Ala-Gln	增强皮肤对紫外线损伤的防御和修护能力以及防止光老化，在防晒护肤品中应用前景广阔	[10-11]
抗癌肽	a.诱导细胞凋亡：通过激活凋亡信号通路，如Caspase级联反应，诱导癌细胞凋亡；b.抑制血管生成：通过抑制血管内皮生长因子（Vascular endothelial growth factor, VEGF）活性，减少肿瘤血管生成，从而抑制肿瘤生长；典型氨基酸序列：Xxx-Arg-Gly-Asp-Xxx	肽易于人体吸收，可选择性地抑制肿瘤细胞生长，同时对正常细胞影响较小，亦可降低化疗药物毒副作用	[12-14,20]
免疫调节肽	a.增强免疫细胞功能：促进T细胞、B细胞和自然杀伤细胞（Natural killer cell, NK Cell）的增殖和活化，提高免疫系统整体功能；b.调节细胞因子分泌：通过调节细胞因子分泌，平衡促炎和抗炎反应，维持免疫稳态；典型氨基酸序列：Gly-Arg-Gly-(Asp)₉	增强机体免疫反应或抑制过度免疫反应，达到免疫平衡效果，在调节免疫方面有良好的应用前景	[15]
美白肽	a.抑制酪氨酸酶活性：美白肽通过与酪氨酸酶结合，抑制其活性，从而减少黑色素的合成；b.干扰黑色素转运：某些美白肽可干扰黑色素小体从黑素细胞向角质形成细胞的转运，减少皮肤色素沉着；典型氨基酸序列： H-Met-Pro-D-Phe-Arg-D-Trp-Phe-Lys-Pro-Val-NH₂	具有更好的安全性和美白活性，可在美白护肤品中广泛应用	[21-22]
减肥肽	a.促进胰岛素分泌：胰高血糖素样肽-1受体激动剂（Glucagon-like peptide-1 receptor agonists, GLP-1RA）可增加胰岛素分泌，降低血糖水平；b.抑制胰高血糖素分泌：GLP-1RA可减少胰高血糖素的分泌，减缓肝脏释放葡萄糖；c.延缓胃排空：GLP-1RA能延缓胃排空速度，使饭后血糖上升更为温和；d.减少食欲：GLP-1RA可降低食欲；典型氨基酸序列：His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly	GLP-1RA不仅能够良好安全的控制血糖水平，适用于2型糖尿病患者的血糖控制；而且能有效减少热量摄入，帮助控制体重，达到有效减肥效果	[23-24]

分类	作用机制和典型氨基酸序列	特点和应用场景	参考文献
抗菌肽	a.破坏细胞膜结构：通过与细菌细胞膜中的磷脂双层结合，增加膜通透性，导致细胞内容物泄漏，从而杀死细菌；b.抑制细菌代谢途径：干扰细菌DNA、RNA和蛋白质合成，抑制细菌生长和繁殖；典型氨基酸序列：Phe-Trp-Lys-Phe-Lys	广谱抗菌、抗感染，特别是对抗耐药性菌株感染有很好效果；伤口愈合；预防感染；女性阴道念珠菌感染等	[3-4]
抗病毒肽	a.阻断病毒进入细胞：通过与病毒外壳蛋白或宿主细胞受体结合，阻止病毒附着和进入宿主细胞；b.抑制病毒复制：干扰病毒RNA或DNA合成，从而抑制病毒复制；典型氨基酸序列：Ala-Ala-Val-Ala-Leu-Leu-Pro-Ala-Val--Leu-Leu-Ala-Leu-Leu-Ala-Pro	通过抑制病毒蛋白的功能，阻断病毒生命周期；可预防病毒感染，对于已感染病毒患者，则可阻止病毒在体内的进一步扩展	[4,19]
抗氧化肽	a.清除自由基：通过捐献电子或氢原子，中和体内自由基，减少自由基对细胞损伤；b.提高抗氧化酶活性：通过提高超氧化物歧化酶（Superoxide dismutase, SOD）和过氧化氢酶等抗氧化酶的活性，增强机体抗氧化能力；典型氨基酸序列：Leu-Ala-Asn-Ala-Lys	安全性高，可以用在食品领域抗氧化上；也可开发相应抗氧化化妆品、保健食品或者药品	[5-9]
抗紫外肽	a.吸收紫外线：通过吸收紫外线光子，减少其对皮肤的穿透和损伤；b.修复紫外线损伤：通过促进DNA修复机制，修复紫外线引起的DNA损伤，减少皮肤癌的发生；典型氨基酸序列：Leu-Val-Asn-Glu-Leu-Thr-Glu-Phe-Ala-Gln	增强皮肤对紫外线损伤的防御和修护能力以及防止光老化，在防晒护肤品中应用前景广阔	[10-11]
抗癌肽	a.诱导细胞凋亡：通过激活凋亡信号通路，如Caspase级联反应，诱导癌细胞凋亡；b.抑制血管生成：通过抑制血管内皮生长因子（Vascular endothelial growth factor, VEGF）活性，减少肿瘤血管生成，从而抑制肿瘤生长；典型氨基酸序列：Xxx-Arg-Gly-Asp-Xxx	肽易于人体吸收，可选择性地抑制肿瘤细胞生长，同时对正常细胞影响较小，亦可降低化疗药物毒副作用	[12-14,20]
免疫调节肽	a.增强免疫细胞功能：促进T细胞、B细胞和自然杀伤细胞（Natural killer cell, NK Cell）的增殖和活化，提高免疫系统整体功能；b.调节细胞因子分泌：通过调节细胞因子分泌，平衡促炎和抗炎反应，维持免疫稳态；典型氨基酸序列：Gly-Arg-Gly-(Asp)₉	增强机体免疫反应或抑制过度免疫反应，达到免疫平衡效果，在调节免疫方面有良好的应用前景	[15]
美白肽	a.抑制酪氨酸酶活性：美白肽通过与酪氨酸酶结合，抑制其活性，从而减少黑色素的合成；b.干扰黑色素转运：某些美白肽可干扰黑色素小体从黑素细胞向角质形成细胞的转运，减少皮肤色素沉着；典型氨基酸序列： H-Met-Pro-D-Phe-Arg-D-Trp-Phe-Lys-Pro-Val-NH₂	具有更好的安全性和美白活性，可在美白护肤品中广泛应用	[21-22]
减肥肽	a.促进胰岛素分泌：胰高血糖素样肽-1受体激动剂（Glucagon-like peptide-1 receptor agonists, GLP-1RA）可增加胰岛素分泌，降低血糖水平；b.抑制胰高血糖素分泌：GLP-1RA可减少胰高血糖素的分泌，减缓肝脏释放葡萄糖；c.延缓胃排空：GLP-1RA能延缓胃排空速度，使饭后血糖上升更为温和；d.减少食欲：GLP-1RA可降低食欲；典型氨基酸序列：His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly	GLP-1RA不仅能够良好安全的控制血糖水平，适用于2型糖尿病患者的血糖控制；而且能有效减少热量摄入，帮助控制体重，达到有效减肥效果	[23-24]

对比维度	固相合成法（SPPS）	液相合成法（LPPS）
原理与过程	固相载体上逐步缩合氨基酸	溶液中氨基酸/肽片段进行耦合
自动化程度	高，流程化的操作可以通过自动化的多肽合成设备实现自动化生产	较低，操作过程较为复杂，目前难以实现大规模自动化生产
适用范围	中长链多肽（通常40个氨基酸以内）	短肽及特定结构要求的多肽（通常10个氨基酸以内）
产率	较高，随着肽链延长收率下降	适中，随着肽链延长收率迅速下降
纯度	较高，随着肽链延长纯度下降	适中，随着肽链延长纯度迅速下降
耗时	较长，通常2-3小时/氨基酸	适中，合成简洁迅速
成本	较高，树脂和大量溶剂	相对较低

对比维度	固相合成法（SPPS）	液相合成法（LPPS）
原理与过程	固相载体上逐步缩合氨基酸	溶液中氨基酸/肽片段进行耦合
自动化程度	高，流程化的操作可以通过自动化的多肽合成设备实现自动化生产	较低，操作过程较为复杂，目前难以实现大规模自动化生产
适用范围	中长链多肽（通常40个氨基酸以内）	短肽及特定结构要求的多肽（通常10个氨基酸以内）
产率	较高，随着肽链延长收率下降	适中，随着肽链延长收率迅速下降
纯度	较高，随着肽链延长纯度下降	适中，随着肽链延长纯度迅速下降
耗时	较长，通常2-3小时/氨基酸	适中，合成简洁迅速
成本	较高，树脂和大量溶剂	相对较低

名称	原料	蛋白酶种类	酶解条件	分离纯化	功能描述	参考文献
降压肽	大豆蛋白	嗜热菌蛋白酶、胃蛋白酶、胰蛋白酶	5% w/v，55°C，pH 8, 3h和37°C，pH 8, 3 h和37°C，pH 7.6，3 h	离心、LC-MS	ACE抑制活性	[45]
降压肽	诃子果实	胃蛋白酶	1:25w/w，37°C，12 h	离心、RP-HPLC	ACE抑制活性	[46]
降压肽	米糠	胰蛋白酶	50 mg/ml，1500 U/mg， 37°C，pH 8，2 h	离心	ACE抑制活性、抗氧化	[47]
降压肽	鸡皮	碱性蛋白酶	1% w/v，60°C，pH 9.5，4 h	离心、超滤	ACE抑制活性	[48]
降压肽	短鳍鱼	碱性蛋白酶	1% w/v	离心、过滤	ACE抑制活性	[49]
抗氧化肽	牛毛	碱性蛋白酶	5%，55°C，8 h	离心、凝胶过滤、分子排阻色谱	抗氧化	[54]
二肽酶抑制肽	三文鱼鱼白	蜂蜜曲霉蛋白酶	5种蛋白酶筛选，50°C，>5 h， 17.5 g酶/7.2 kg原料	硅藻土过滤、超滤、浓缩	控制血糖	[55]
血糖控制以及抗炎肽	去骨三文鱼	胃蛋白酶、胰蛋白酶、胰凝乳蛋白酶	N/A	过滤、超滤	控制血糖、抗炎	[56]
抗氧化肽	黄鳍金枪鱼	碱性蛋白酶	酶量3000 U/g，50°C，5 h，100 rpm	离心、超滤 Sephadex G-15	抗氧化	[57]
抗氧化肽	鱿鱼	碱性蛋白酶	酶量5-30 U/g，5-180 min	离心	抗氧化	[58]
抗癌肽	虾壳	Cryotin酶	pH8，50°C，45 min	离心、超滤	抗恶性细胞增殖	[59]