Article(id=1153986714575032423, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986709126635984, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241105001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1730736000000, receivedDateStr=2024-11-05, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1753061472791, onlineDateStr=2025-07-21, pubDate=1737734400000, pubDateStr=2025-01-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1753061472791, onlineIssueDateStr=2025-07-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1753061472791, creator=13701087609, updateTime=1753061472791, updator=13701087609, issue=Issue{id=1153986709126635984, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='2', pageStart='1', pageEnd='324', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1753061471492, creator=13701087609, updateTime=1760345674980, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1184538872999457117, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986709126635984, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1184538872999457118, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986709126635984, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=231, endPage=238, ext={EN=ArticleExt(id=1153986715271286890, articleId=1153986714575032423, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Research progress on the isolation and purification of active peptides in fermented soybean foods and their hypoglycaemic mechanisms, columnId=1151923894560060071, journalTitle=Journal of Food Safety & Quality, columnName=Food Nutrition and Functional Foods, runingTitle=null, highlight=null, articleAbstract=

In recent years, bioactive peptides with potential edible values have received widespread attention as consumer demand for functional foods grows. The bioactive peptides in fermented soybean foods (FSF) have attracted much attention because of their safe and non-toxic properties. Nowadays, a large number of FSF peptides have been identified through isolation and purification, which not only contain rich nutritional value, but also have a variety of physiological activities such as hypoglycemia, hypotension, antioxidant and antimicrobial activities, and have a good prospect for development in the field of food and health care products. This paper systematically classified and analyzed the biological activities of the active peptides in FSF in the light of the research progress of FSF in recent years, summarized the separation and purification methods of FSF peptides, and summarized the hypoglycemic mechanism of the active peptides in FSF, in order to provide ideas for the deeper study of FSF, and to provide theoretical foundations and support for the production of FSF active peptides and their development and application in the field of functional foods in the future.

, correspAuthors=Na ZHANG, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Man GAO, Yang YANG, Chun-Min MA, Lian-Zhong AI, Feng-Xiang LIN, Yu-Xin WANG, Xiu-Jun ZHANG, Hui-Min ZHANG, Hai-Yan WANG, Na ZHANG), CN=ArticleExt(id=1153986743029191580, articleId=1153986714575032423, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=发酵豆制品中活性肽的分离纯化及其降血糖机制研究进展, columnId=1151923894698472105, journalTitle=食品安全质量检测学报, columnName=食品营养及功能性食品, runingTitle=null, highlight=null, articleAbstract=

近年来, 随着消费者对功能性食品需求的日益增加, 具有潜在食用价值的生物活性肽受到了广泛的关注。特别是发酵豆制品(fermented soybean foods, FSF)中的生物活性肽, 因其安全无毒的特性而备受瞩目。目前, 大量的FSF活性肽经过分离和纯化已被鉴定。FSF活性肽不仅含有丰富的营养价值, 还具有降血糖、降血压、抗氧化和抗菌等多种生理活性, 在食品和保健品等领域发展前景良好。本文结合近年来FSF的研究进展, 系统归类和分析了FSF中活性肽的生物活性, 总结了FSF生物活性肽分离纯化方法, 并归纳了FSF中活性肽的降血糖机制, 以期为FSF更深层次的研究提供思路, 为未来FSF活性肽的生产及其在功能性食品领域中的开发应用提供理论基础和支撑。

, correspAuthors=张娜, authorNote=null, correspAuthorsNote=
* 张娜(1979—), 女, 博士, 教授, 主要研究方向为谷物与粮食加工。E-mail:
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高嫚(1999—), 女, 博士研究生, 主要研究方向为谷物化学利用与研究。E-mail:

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高嫚(1999—), 女, 博士研究生, 主要研究方向为谷物化学利用与研究。E-mail:

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高嫚(1999—), 女, 博士研究生, 主要研究方向为谷物化学利用与研究。E-mail:

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注: A. 超滤; B. 离子交换色谱; C. 凝胶过滤色谱; D. 反相高效液相色谱。

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注: 二肽基肽酶Ⅳ (dipeptidyl peptidase IV, DPP-Ⅳ); 胰高血糖素样肽-1 (glucagon-like peptide-1, GLP-1)。

, figureFileSmall=4z6CYqoh+E2RWSoSBjJ4pQ==, figureFileBig=9HX/xaZP014rrhrh9a3j8w==, tableContent=null), ArticleFig(id=1184566965252277041, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986714575032423, language=EN, label=Table 1, caption=

Methods for identification and characterization of FSF active peptides

, figureFileSmall=null, figureFileBig=null, tableContent=
来源 鉴定方法 肽序列 生物活性 文献
郫县蚕豆酱 LC-MS RGLSK、NKGPR、DNLLN和TPCPPQ 降血压、抗氧化 [26]
发酵蚕豆酱 MS LY4、LP-5和VL-9 抗氧化、抗菌 [27]
豆豉 MS VY和SFLLR 降血糖 [50]
无盐酱油 MS Ala-Phe和Ile-Phe 降血压 [51]
发酵油腐乳 LC-MS FALP、GWPT和EGWP 降血压 [52]
发酵豆奶 LC-MS/MS 肽序列已知, 其中16种为降血压肽, 18种为抗氧化肽 降血压、抗氧化 [53]
), ArticleFig(id=1184566965357134643, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986714575032423, language=CN, label=表1, caption=

FSF活性肽鉴定与表征方法

, figureFileSmall=null, figureFileBig=null, tableContent=
来源 鉴定方法 肽序列 生物活性 文献
郫县蚕豆酱 LC-MS RGLSK、NKGPR、DNLLN和TPCPPQ 降血压、抗氧化 [26]
发酵蚕豆酱 MS LY4、LP-5和VL-9 抗氧化、抗菌 [27]
豆豉 MS VY和SFLLR 降血糖 [50]
无盐酱油 MS Ala-Phe和Ile-Phe 降血压 [51]
发酵油腐乳 LC-MS FALP、GWPT和EGWP 降血压 [52]
发酵豆奶 LC-MS/MS 肽序列已知, 其中16种为降血压肽, 18种为抗氧化肽 降血压、抗氧化 [53]
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发酵豆制品中活性肽的分离纯化及其降血糖机制研究进展
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高嫚 1 , 杨杨 1 , 马春敏 1 , 艾连中 2 , 林枫翔 3 , 王玉鑫 4 , 张修军 5 , 张慧敏 6 , 王海燕 7 , 张娜 1, *
食品安全质量检测学报 | 食品营养及功能性食品 2025,16(2): 231-238
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食品安全质量检测学报 | 食品营养及功能性食品 2025, 16(2): 231-238
发酵豆制品中活性肽的分离纯化及其降血糖机制研究进展
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高嫚1 , 杨杨1, 马春敏1, 艾连中2, 林枫翔3, 王玉鑫4, 张修军5, 张慧敏6, 王海燕7, 张娜1, *
作者信息
  • 1.哈尔滨商业大学食品工程学院, 哈尔滨 150028
  • 2.上海理工大学健康科学与工程学院, 上海, 200093
  • 3.哈尔滨美华生物技术股份有限公司, 哈尔滨 150010
  • 4.哈尔滨市太阳岛调味食品股份有限公司, 哈尔滨 150001
  • 5.黑龙江九阳豆业有限公司, 哈尔滨 161542
  • 6.黑龙江省克东腐乳有限公司, 齐齐哈尔 164800
  • 7.哈尔滨北大荒豆制品有限公司, 哈尔滨 150008
  • 高嫚(1999—), 女, 博士研究生, 主要研究方向为谷物化学利用与研究。E-mail:

通讯作者:

* 张娜(1979—), 女, 博士, 教授, 主要研究方向为谷物与粮食加工。E-mail:
Research progress on the isolation and purification of active peptides in fermented soybean foods and their hypoglycaemic mechanisms
Man GAO1 , Yang YANG1, Chun-Min MA1, Lian-Zhong AI2, Feng-Xiang LIN3, Yu-Xin WANG4, Xiu-Jun ZHANG5, Hui-Min ZHANG6, Hai-Yan WANG7, Na ZHANG1, *
Affiliations
  • 1. College of Food Engineering, Harbin University of Commerce, Harbin 150028, China
  • 2. School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China
  • 3. Harbin Meihua Biotechnology Co., Ltd., Harbin 150010, China
  • 4. Harbin Taiyangdao Seasoning Food Co., Ltd., Harbin 150001, China
  • 5. Heilongjiang Joyang Soybean Industry Co., Ltd., Harbin 161542, China
  • 6. Heilongjiang Kedong Fermented Bean Curd Co., Ltd., Qiqihar 164800, China
  • 7. Harbin Beidahuang Soybean Products Co., Ltd., Harbin 150008, China
出版时间: 2025-01-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241105001
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近年来, 随着消费者对功能性食品需求的日益增加, 具有潜在食用价值的生物活性肽受到了广泛的关注。特别是发酵豆制品(fermented soybean foods, FSF)中的生物活性肽, 因其安全无毒的特性而备受瞩目。目前, 大量的FSF活性肽经过分离和纯化已被鉴定。FSF活性肽不仅含有丰富的营养价值, 还具有降血糖、降血压、抗氧化和抗菌等多种生理活性, 在食品和保健品等领域发展前景良好。本文结合近年来FSF的研究进展, 系统归类和分析了FSF中活性肽的生物活性, 总结了FSF生物活性肽分离纯化方法, 并归纳了FSF中活性肽的降血糖机制, 以期为FSF更深层次的研究提供思路, 为未来FSF活性肽的生产及其在功能性食品领域中的开发应用提供理论基础和支撑。

发酵豆制品  /  生物活性肽  /  分离纯化  /  降血糖

In recent years, bioactive peptides with potential edible values have received widespread attention as consumer demand for functional foods grows. The bioactive peptides in fermented soybean foods (FSF) have attracted much attention because of their safe and non-toxic properties. Nowadays, a large number of FSF peptides have been identified through isolation and purification, which not only contain rich nutritional value, but also have a variety of physiological activities such as hypoglycemia, hypotension, antioxidant and antimicrobial activities, and have a good prospect for development in the field of food and health care products. This paper systematically classified and analyzed the biological activities of the active peptides in FSF in the light of the research progress of FSF in recent years, summarized the separation and purification methods of FSF peptides, and summarized the hypoglycemic mechanism of the active peptides in FSF, in order to provide ideas for the deeper study of FSF, and to provide theoretical foundations and support for the production of FSF active peptides and their development and application in the field of functional foods in the future.

fermented soybean foods  /  bioactive peptides  /  isolation and purification  /  hypoglycaemia
高嫚, 杨杨, 马春敏, 艾连中, 林枫翔, 王玉鑫, 张修军, 张慧敏, 王海燕, 张娜. 发酵豆制品中活性肽的分离纯化及其降血糖机制研究进展. 食品安全质量检测学报, 2025 , 16 (2) : 231 -238 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241105001
Man GAO, Yang YANG, Chun-Min MA, Lian-Zhong AI, Feng-Xiang LIN, Yu-Xin WANG, Xiu-Jun ZHANG, Hui-Min ZHANG, Hai-Yan WANG, Na ZHANG. Research progress on the isolation and purification of active peptides in fermented soybean foods and their hypoglycaemic mechanisms[J]. Journal of Food Safety & Quality, 2025 , 16 (2) : 231 -238 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241105001
发酵豆制品(fermented soybean foods, FSF), 是指以大豆、大豆制品及其加工副产物为发酵基质, 在一种或多种微生物作用下制备具有特定风味和形态的食品或调味品[1]。由于其丰富的营养成分和生物活性成分, 如氨基酸、纤维、钙、钾、异黄酮、大豆皂苷和木脂素, FSF受到亚洲乃至全球消费者的青睐[2-3]。与生大豆相比, FSF含有更丰富的膳食成分, 这些成分具有更长的保质期、更佳的感官体验、更高的消化率和更优的功能特性[4]。近年来, 随着生物技术的迅速发展, FSF中有关活性成分的研究, 特别是活性肽的研究, 已成为食品科学领域的研究热点之一。
生物活性肽是由多个氨基酸残基组成的短链肽, 它们通过血液循环参与生物体的生理调节[5]。研究指出, 在治疗人类慢性疾病(图1)方面, 生物活性肽的效果比大分子蛋白质和小分子氨基酸更为显著[6]。富含蛋白质的食品原料是制备生物活性肽的重要来源。FSF作为日常饮食中极佳的植物蛋白来源, 经过微生物发酵后, 有效提升了大豆生物活性肽的含量[7]。近年来, 科研人员通过采用不同的提取技术, 成功从FSF蛋白质中分离出具有特定功效的生物活性肽, 如降血糖肽、降血压肽、抗氧化肽和抗菌肽等, 显著提升了大豆的附加值[8-9]
在众多生物活性肽中, 降血糖肽因其对血糖水平的调节作用而备受关注。糖尿病作为一种全球性的慢性疾病, 患者人数不断攀升, 对人们的健康和生活质量构成了严重威胁。传统的治疗方法虽然取得了一定的效果, 但副作用和长期依赖性等问题仍然存在[10]。因此, 寻找安全、有效的降血糖成分成为了当前研究的重点。FSF中的降血糖肽作为一种天然、无副作用的潜在治疗成分, 具有重要的研究价值和应用前景。
本文对近年来FSF中活性肽的分离纯化技术及其降血糖机制进行了综述。首先概述了FSF中活性肽的种类及其在预防和治疗慢性疾病方面的潜在应用, 详细探讨了FSF中活性肽的分离纯化技术, 涵盖了多种制备方法、纯化流程以及鉴定技术。此外还分析了降血糖肽的作用机制, 并对FSF中活性肽研究的未来趋势进行了展望, 以期为FSF中活性肽的进一步研究和应用提供科学依据和参考。
FSF作为一种在许多亚洲国家广受欢迎的食品, 是获取生物活性肽的一个重要来源。FSF中肽的来源主要有两种途径。(1)大豆蛋白在发酵过程中水解释放形成, 在大豆发酵过程中, 其蛋白成分会被水解成具有特定生物活性的肽。(2)由参与发酵的微生物产生。不同的微生物菌株在发酵中能促成特定生物活性肽的形成, 这些肽依据其氨基酸的序列和构成, 表现出不同的生物活性, 如降血糖、降血压、抗氧化、抗菌等[11]
到2045年, 预计全球将有7.83亿人受到糖尿病的影响, 其中大约80%的人口生活在中低收入国家[12]。长期的高血糖水平会增加多种疾病的风险, 包括失明、神经病变和心血管疾病[13]。将血糖控制在正常范围内是减缓甚至预防糖尿病及其相关并发症的有效手段。尽管胰岛素制剂能够帮助调节血糖水平, 但其高昂的成本可能给糖尿病患者带来沉重的经济压力。虽然存在一些价格合理的合成药物, 如米格列醇和伏格列波糖, 但这些药物通常伴随着不良副作用[14]。因此, 迫切需要开发既经济实惠又副作用小的新型降糖成分。在FSF中的生物活性肽, 特别是降血糖肽, 因其潜在的健康益处而受到广泛关注。YU等[15]从豆豉中分离鉴定得到的两种肽VY和SFLLR, 可以改善L6细胞的葡萄糖摄取能力, 具有降血糖的潜力。此外, 在另一项研究中, YU等[16]发现这两种肽都可以改善高脂肪饮食诱导的胰岛素抵抗小鼠的葡萄糖稳态, 并缓解肠道菌群失调。为了进一步探索降血糖肽的潜力, 研究者们开始关注其在不同FSF中的存在和作用。从纳豆中提取的生物活性肽被发现能够抑制α-葡萄糖苷酶的活性, 从而减缓餐后血糖的上升速度[17]。此外, 从发酵豆腐中分离出的特定肽段也显示出对α-淀粉酶的抑制作用, 有助于降低餐后血糖水平[18]。这些研究结果表明, FSF中的降血糖肽具有成为糖尿病辅助治疗手段的潜力。
高血压是全球最普遍的心血管疾病, 正日益成为一个严重的问题。世界卫生组织推荐的降压药物包括噻嗪类药物、血管紧张素转换酶(angiotensin converting enzyme, ACE)抑制剂、血管紧张素受体阻滞剂以及长效二氢吡啶钙通道阻滞剂[19]。这些药物能够有效地降低血压, 但它们具有多种副作用, 例如血脂升高、脚踝肿胀、咳嗽和眩晕。作为降压药物的潜在有效替代品, 食物来源的降血压肽由于其特异性和安全性受到了研究人员的广泛关注。
降血压肽存在于FSF中, 已被证明可以降低血压。这些肽可以通过人体消化、循环和神经系统的多种机制发挥其降血压作用[20]。降血压肽起作用的主要机制之一是抑制ACE。ACE通过将血管紧张素I转化为血管紧张素II在血压调节中发挥关键作用[21]。降血压肽可以阻断这种转化, 导致血管舒张和血压降低。这些肽的另一种机制是刺激NO的产生, NO是一种可放松血管和促进血管舒张的分子。NO水平升高会导致血压降低。GUO等[22]研究了豆豉提取物对ACE的影响, 发现豆豉提取物对ACE的平均抑制率为4.56%~27.07%。并且, 他们利用肽组学和分子对接相结合的方法鉴定出了4种具有潜在ACE抑制活性的新型肽: PSSPFTDLWD、EEQDERQFPF、PVPVQQAFPF和PSSPFTDL。SOANKI等[23]采用发酵乳杆菌(M2)和干酪乳杆菌(NK9)进行豆奶发酵, 结果表明由M2发酵的豆奶与NK9相比具有更高的ACE抑制活性, 这表明不同菌株发酵产生的活性肽具有不同的降血压效果。除了豆豉和发酵豆奶, 其他FSF如腐乳和豆酱也被发现含有降血压肽。降血压肽可制成药片, 也可作为功能性成分添加到食品中, 具有良好的市场应用潜力和经济效益。
自由基过量累积会导致人体许多慢性疾病的产生, 如衰老、慢性炎症、心血管疾病等, 目前常规的解决方法是使用抗氧化剂。然而人工合成的抗氧化剂对人体有潜在危害, 因此无毒副作用的抗氧化肽成为了当今研究的热点。目前, 抗氧化肽的主要来源包括动物蛋白、植物蛋白、微生物蛋白等[24]。其中, FSF富含丰富的蛋白质, 被认为是天然抗氧化肽的一个重要来源。FSF抗氧化肽通过清除机体内的活性氧和自由基, 干扰氧化链式反应, 发挥抗氧化的作用[25]。LI等[26]于2021年首次从郫县蚕豆酱中发现了四种新型生物活性肽, 其中TPCPPQ表现出最高的抗氧化活性, 证明郫县蚕豆酱是抗氧化肽的良好来源。LIN等[27]基于计算机模拟和体外分析, 鉴定并合成了7种源自郫县蚕豆酱的新型抗氧化肽, 具有显著的抗氧化能力。ZHANG等[28]研究发现, 从豆鼓中鉴定出的3种肽EEQEHT、FED、AEPE具有较强的清除自由基的能力, 抗氧化活性较好。此外, 发酵豆腐中的某些肽也被证明具有抑制脂质过氧化的作用, 从而有助于预防与氧化应激相关的疾病。这些发现表明, FSF中的活性肽在抗氧化方面具有巨大的潜力, 未来的研究可以进一步探讨其在预防和治疗相关疾病中的应用。
由有害微生物引起的食源性感染已成为一个全球性问题, 严重危害人类健康。根据世界卫生组织统计, 全球约10%的人口因食用受污染的食物而被感染, 每年有近42万人死于食源性疾病[29]。尽管化学食品防腐剂可有效抑制病原菌生长和延长食品保质期, 但消费者对其安全性的担忧仍然存在。因此, 为了提高食品安全性和延长保质期, 有必要探索传统防腐剂的天然替代品。抗菌肽作为一种在食品工业中广泛应用的天然防腐剂, 具有优异的广谱抗菌性能, 同时对哺乳动物具有低毒性, 且不易诱导耐药性[30]。目前, 已在多种FSF中鉴定出抗菌肽。YEO等[31]从枯草芽孢杆菌SCK-2发酵的大豆酱中成功分离出具有抗菌活性的肽, 这些抗菌肽对蜡样芽孢杆菌增殖具有显著的抑制作用。KITAGAWA等[32]从纳豆中分离出一种含45个的氨基酸残基的肽, 该肽对肺炎链球菌、枯草芽孢杆菌、短芽孢杆菌和地衣芽孢杆菌的生长繁殖均具有抑制作用。这一发现进一步证实了FSF作为抗菌肽丰富来源的潜力, 并为食品工业中的微生物控制提供了新的思路。
图2所示, 目前从FSF中获取生物活性肽的途径主要有3种[33]。第一种是直接提取法, 即从FSF中直接萃取微生物自然产生的内源性生物活性肽。直接提取法常常和一些高新技术例如超声、微波和高静水压力等联用。这些新技术作为辅助手段用于提取生物活性肽, 能够加速多肽的释放, 促进其迅速溶解于溶剂中[34]。在特定条件下, 这些方法不会破坏多肽的结构, 并且操作简便, 能够节省时间并提升工作效率。第二种是酶解法, 这也是获取生物活性肽的重要手段。通过向FSF或其分离蛋白中添加特定的外源酶, 如胰蛋白酶、胃蛋白酶或木瓜蛋白酶等, 可以有效地分解蛋白质, 释放出具有生物活性的肽片段[35]。这种方法的优势在于可以通过选择不同的酶来定向获得特定功能的活性肽, 例如降血糖肽、降血压肽或抗氧化肽等。此外, 酶解法还可以在一定程度上模拟人体内的消化过程, 有助于研究活性肽在消化系统中的稳定性及其生物利用度[36]。除了直接提取法和酶解法, 分析预测法也可以用于FSF中活性肽的制备。分析预测法是基于肽数据库和多种生物信息学工具, 通过分析已知的活性肽序列和结构, 预测新的潜在活性肽[37]。这种方法可以大大减少实验工作量, 提高活性肽筛选的效率和准确性。通过这种方法, 研究人员可以快速识别出具有特定功能的肽序列, 并通过合成或基因工程手段进行验证和应用开发。随着生物信息学和计算生物学的不断发展, 分析预测法在FSF活性肽研究中的应用前景将更加广阔。
选择合适的分离技术能够更好地分离纯化粗多肽, 从而得到高活性、高纯度的活性肽, 这是分析FSF活性肽结构与活性的基础[38]。在FSF活性肽的分离纯化过程中, 常用的分离技术为膜分离技术和色谱技术[39]。下面对FSF生物活性肽的分离纯化过程进行了汇总, 结果见图3
膜分离技术利用膜两侧的压力差来截留和分离不同分子量的物质。根据膜孔径的不同, 该技术主要分为微滤、超滤、纳滤和反渗透4种类型[40]。超滤技术因其操作的快速性和简便性, 在FSF活性肽的纯化过程中得到了广泛的应用。通过施加压力, FSF蛋白水解产物被推动通过超滤膜, 其中小分子肽能够穿过膜上的孔隙, 从而实现小分子肽与大分子蛋白的有效分离[41]。此外, 不溶性物质和可溶性杂质也能与FSF活性肽初步分离。同时, 采用不同孔隙尺寸的超滤膜, 可以实现不同分子量活性肽的分离[42]。然而, 用超滤技术很难有效分离相对分子质量很小或相似的多肽。因此, 在实际研究中, 常与色谱技术相结合。
常用的色谱分离方法包括: 离子交换色谱、凝胶过滤色谱和反相高效液相色谱。离子交换色谱主要利用不同物质在离子交换树脂上的吸附能力差异进行分离[43]。通过调节pH和离子强度, 可以实现不同电荷状态的FSF活性肽的分离。凝胶过滤色谱则依据分子大小和形状的不同, 通过凝胶介质的孔隙进行分离[44]。小分子物质能够进入凝胶孔隙, 而大分子物质则被排除在外, 从而实现分离。反相高效液相色谱则通过调节流动相的极性, 利用疏水作用力实现不同疏水性的FSF活性肽的分离[45]。这些色谱技术在分离纯化过程中具有高效、高分辨率的特点, 能够进一步提高FSF活性肽的纯度和质量。其中, 反相高效液相色谱法由于其稳定性好、分离效率高、应用范围广, 已被广泛应用于FSF活性肽的分离纯化。
值得注意的是, 上述几种活性肽分离纯化技术各有优缺点, 单一的分离纯化技术并不能完全满足FSF活性肽高效分离纯化的要求[46]。故在FSF活性肽分离纯化的实际研究中, 常常综合考虑FSF活性肽的多种性质, 采用多种分离技术进行分离纯化, 以达到更好的分离纯化效果[47]
从FSF中分离出的具有潜在生物活性的肽需要通过质谱法进行鉴定并人工合成以验证活性[48]。质谱法通过测定分子离子的质量/电荷比, 可以确定肽段的分子量和结构信息。在FSF活性肽鉴定过程中, 质谱法通常与液相色谱技术联用, 形成液相色谱-质谱法(liquid chromatography- mass spectrometry, LC-MS), 以提高鉴定的灵敏度和准确性。通过LC-MS分析, 可以得到肽段的精确分子量、氨基酸序列以及可能的修饰信息。除了MS, FSF活性肽的鉴定还可以通过其他方法进行, 如核磁共振技术。核磁共振谱法能够提供分子结构的详细信息, 特别是对于三维结构的解析非常有用。此外, FSF活性肽的鉴定还可以借助于生物信息学工具, 通过数据库比对和预测分析来辅助确定肽段的序列和功能[49]
表1为不同来源FSF的活性肽鉴定与表征方法。在FSF活性肽表征方面, 除了鉴定其结构外, 还需要对其生物活性进行评估。这通常涉及到一系列体外和体内实验, 如酶抑制实验、细胞活性实验和动物模型实验等。通过这些实验可以验证FSF活性肽的降血糖、降血压、抗氧化和抗菌等生物活性, 为进一步的机制研究和应用开发奠定基础。
图4所示, FSF中降血糖肽主要通过抑制α-淀粉酶、α-葡萄糖苷酶以及二肽基肽酶Ⅳ(dipeptidyl peptidase IV, DPP-Ⅳ)的活性来发挥作用, 有效降低血清葡萄糖水平, 进而实现治疗糖尿病的目标[55]
α-淀粉酶能够水解淀粉中的α-1,4-糖苷键和α-1,6-糖苷键, 将其分解为糊精、寡糖等中间产物。这些中间产物在小肠中进一步被α-葡萄糖苷酶水解为单糖(如葡萄糖), 供人体吸收利用。α-淀粉酶的活性直接影响餐后血糖的升高幅度。当α-淀粉酶活性较高时, 淀粉等碳水化合物的消化速度加快, 导致餐后血糖迅速升高[56]。反之, 当α-淀粉酶活性受到抑制时, 淀粉的消化速度减慢, 餐后血糖的升高幅度也会相应降低[57]。降血糖肽中的特定成分能够与α-淀粉酶的活性位点结合, 从而降低其催化淀粉水解的速率。这种抑制作用减缓了淀粉分解为葡萄糖的过程, 进而降低了餐后血糖的升高幅度。
α-葡萄糖苷酶是小肠内的一种酶, 主要负责将多糖分解为单糖, 从而促进葡萄糖的吸收。降血糖肽通过竞争性抑制α-葡萄糖苷酶的活性, 减缓了糖类物质的分解速度, 从而降低了餐后血糖的上升速度[58]。研究表明, 某些特定的活性肽序列能够与α-葡萄糖苷酶的活性位点结合, 阻碍底物的进入, 从而抑制其催化作用[59]。这种抑制作用具有可逆性, 因此在不影响正常糖类代谢的前提下, 有效控制血糖水平。
降血糖肽还可以通过抑制DPP-Ⅳ的活性来发挥其降血糖作用。DPP-Ⅳ是一种膜结合的丝氨酸蛋白酶, 主要负责降解肠促胰岛素激素[60], 如GLP-1和葡萄糖依赖性促胰岛素多肽(glucose-dependent insulinotropic polypeptide, GIP)。这些肠促胰岛素激素在调节血糖水平方面起着重要作用, 通过促进胰岛素的分泌和抑制胰高血糖素的释放来降低血糖。降血糖肽通过与DPP-Ⅳ的活性位点结合, 抑制其酶活性, 从而延长了GLP-1和GIP的半衰期, 增强了它们的降血糖效果[61]。这种机制不仅有助于降低餐后血糖水平, 还能改善胰岛β细胞的功能, 从而在糖尿病的治疗中发挥重要作用。
除了通过抑制α-淀粉酶、α-葡萄糖苷酶和DPP-Ⅳ的活性来降低血糖水平外, FSF中的降血糖肽还可能通过影响钠-葡萄糖转运蛋白的活性来发挥作用。综上, FSF降血糖肽通过多种机制实现其降血糖效果, 这些机制共同作用, 有助于控制血糖水平, 为糖尿病的治疗提供了新的思路和潜在的治疗策略。
随着对FSF中活性肽研究的不断深入, 已经认识到这些生物活性肽在调节血糖水平、血压、抗氧化和抗菌等方面具有显著的潜力。特别是降血糖肽, 它们通过多种机制协同作用, 为糖尿病的治疗提供了新的思路。未来的研究应着重于活性肽的结构与其生物活性之间的关系, 以及它们在人体内的代谢途径和安全性评估。这将有助于开发出更加高效、安全的新型降血糖药物, 为糖尿病患者带来更多的治疗选择和希望。
随着现代生物技术的不断发展, 利用基因工程和合成生物学等手段, 有望在更短的时间内获得具有特定功能的活性肽。FSF活性肽因其独特的生理功能, 在功能性食品开发中成为理想原料, 能科学配比生产特定健康需求的食品, 同时作为天然防腐剂、抗氧化剂、营养强化剂及食品改良剂提升食品品质, 并在生物医药行业中展现出药物开发、生物材料制备及高灵敏度诊断试剂开发等多重应用前景。通过优化发酵工艺和分离纯化技术, 可以提高活性肽的产量和纯度, 降低生产成本, 使这些有益于健康的活性肽能够更广泛地应用于食品和医药行业。总之, FSF中的活性肽作为一种天然、安全的生物资源, 具有广阔的应用前景。
  • 国家重点研发计划项目(2023YFD2100803)
  • 国家自然科学基金项目(32372387)
  • 国家自然科学基金项目(32402015)
  • 黑龙江省“双一流”学科协同创新成果项目(LJGXCG202080)
  • 黑龙江省“双一流”学科协同创新成果项目(LJGXCG202083)
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2025年第16卷第2期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241105001
  • 接收时间:2024-11-05
  • 首发时间:2025-07-21
  • 出版时间:2025-01-25
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  • 收稿日期:2024-11-05
基金
国家重点研发计划项目(2023YFD2100803)
国家自然科学基金项目(32372387)
国家自然科学基金项目(32402015)
黑龙江省“双一流”学科协同创新成果项目(LJGXCG202080)
黑龙江省“双一流”学科协同创新成果项目(LJGXCG202083)
作者信息
    1.哈尔滨商业大学食品工程学院, 哈尔滨 150028
    2.上海理工大学健康科学与工程学院, 上海, 200093
    3.哈尔滨美华生物技术股份有限公司, 哈尔滨 150010
    4.哈尔滨市太阳岛调味食品股份有限公司, 哈尔滨 150001
    5.黑龙江九阳豆业有限公司, 哈尔滨 161542
    6.黑龙江省克东腐乳有限公司, 齐齐哈尔 164800
    7.哈尔滨北大荒豆制品有限公司, 哈尔滨 150008

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* 张娜(1979—), 女, 博士, 教授, 主要研究方向为谷物与粮食加工。E-mail:
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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