Article(id=1151881500787569585, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151881493552394994, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241204004, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1733241600000, receivedDateStr=2024-12-04, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752559550710, onlineDateStr=2025-07-15, pubDate=1748102400000, pubDateStr=2025-05-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752559550710, onlineIssueDateStr=2025-07-15, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752559550710, creator=13701087609, updateTime=1752559550710, updator=13701087609, issue=Issue{id=1151881493552394994, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='10', 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=1752559548986, creator=13701087609, updateTime=1756202008453, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1167159075906265916, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151881493552394994, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1167159075906265917, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151881493552394994, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=302, endPage=308, ext={EN=ArticleExt(id=1151923899232039718, articleId=1151881500787569585, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Study on the antioxidant activity and stability of peptides from the visceral of Todarodes pacificus, columnId=1151923894560060071, journalTitle=Journal of Food Safety & Quality, columnName=Food Nutrition and Functional Foods, runingTitle=null, highlight=null, articleAbstract=

Objective To explore the antioxidant activity and stability of peptides from the visceral of Todarodes pacificus. Methods Free radical scavenging capacities of peptides from the visceral of Todarodes pacificus were determined, while the effects of temperature, pH and gastrointestinal digestion on the antioxidant activity of peptides from the visceral of Todarodes pacificus were also evaluated. Results The peptides from the visceral of Todarodes pacificus exhibited excellent free radical scavenging abilities, with half maximal inhibition concentration (IC50) values of 1.38, 15.26, 0.90 and 2.21 mg/mL for scavenging 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), 2,2’-azino-bis(3- ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) cationic radicals, hydroxyl radicals, and superoxide anion radicals, respectively. The antioxidant activity of the peptides remained relatively stable within a temperature range of 25-100 °C. Under pH conditions of 7.0-8.0, their antioxidant activity showed no significant change (P>0.05), but it significantly decreased under strongly acidic or alkaline conditions (P<0.05). After in vitro simulated gastrointestinal digestion, the peptides still maintained strong antioxidant activity. Conclusion The peptides from the visceral of Todarodes pacificus has good antioxidant activity and stability, but it shall be noted that the processing and storage of its related products avoid the influence of high temperature and extreme acid-base conditions. The present research will open up new ways for the high-value utilization of the visceral of Todarodes pacificus resources and provide scientific guidance for the innovative development of natural antioxidant peptides.

, correspAuthors=Kai-Liang LENG, 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=Xiao-Fang LIU, Xiang-Yang LI, Xiao-Yan FU, Fu-Hou LI, Wei-Xia WANG, Kai-Liang LENG), CN=ArticleExt(id=1151923920341971616, articleId=1151881500787569585, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=太平洋褶柔鱼内脏蛋白肽的抗氧化活性及稳定性研究, 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=

目的 分析探究太平洋褶柔鱼(Todarodes pacificus)内脏蛋白肽的抗氧化活性及稳定性。方法 测定太平洋褶柔鱼内脏蛋白肽的自由基清除能力, 评价其抗氧化活性, 并考察温度、酸碱、胃肠道消化等因素对其稳定性的影响。结果 太平洋褶柔鱼内脏蛋白肽具有良好的自由基清除能力, 其清除1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl radical, DPPH)自由基、2,2-联氮-2(3-乙基-苯并噻唑-6-磺酸)二铵盐[2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, ABTS]阳离子自由基、羟自由基、超氧阴离子自由基的半数抑制浓度(half maximal inhibition concentration, IC50)分别为1.38、15.26、0.90、2.21 mg/mL。该蛋白肽的抗氧化活性在25~100 ℃温度范围内较为稳定; 在pH 7.0~8.0条件下其抗氧化活性变化不显著(P>0.05), 但在强酸或强碱条件下其抗氧化活性显著下降(P<0.05); 经体外模拟胃肠道消化后, 该蛋白肽依然能够维持良好的抗氧化活性。结论 太平洋褶柔鱼内脏蛋白肽具备良好的抗氧化活性与稳定性, 但在其加工与贮藏过程中还应注意避免高温与极端酸碱条件的影响。研究成果可为太平洋褶柔鱼内脏资源的高值化利用开辟新途径, 也可为天然抗氧化肽的开发提供科学指导。

, correspAuthors=冷凯良, authorNote=null, correspAuthorsNote=
* 冷凯良(1966—), 男, 研究员, 主要研究方向为水产品加工与利用。E-mail:
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刘小芳(1987—), 女, 博士, 副研究员, 主要研究方向为水产品加工与利用。E-mail:

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刘小芳(1987—), 女, 博士, 副研究员, 主要研究方向为水产品加工与利用。E-mail:

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注: 不同字母标识表示不同实验组间具有显著性差异(P<0.05), 下同。

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太平洋褶柔鱼内脏蛋白肽的抗氧化活性及稳定性研究
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刘小芳 1 , 李向阳 1, 2 , 付晓艳 1, 2 , 李福后 2 , 王伟霞 2 , 冷凯良 1, 3, *
食品安全质量检测学报 | 食品营养及功能性食品 2025,16(10): 302-308
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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(10): 302-308
太平洋褶柔鱼内脏蛋白肽的抗氧化活性及稳定性研究
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刘小芳1 , 李向阳1, 2, 付晓艳1, 2, 李福后2, 王伟霞2, 冷凯良1, 3, *
作者信息
  • 1.中国水产科学研究院黄海水产研究所, 农业农村部极地渔业可持续利用重点实验室, 青岛 266071
  • 2.江苏海洋大学海洋食品与生物工程学院, 江苏省海洋生物资源与环境重点实验室, 连云港 222005
  • 3.青岛海洋科技中心, 海洋药物与生物制品功能实验室, 青岛 266237
  • 刘小芳(1987—), 女, 博士, 副研究员, 主要研究方向为水产品加工与利用。E-mail:

通讯作者:

* 冷凯良(1966—), 男, 研究员, 主要研究方向为水产品加工与利用。E-mail:
Study on the antioxidant activity and stability of peptides from the visceral of Todarodes pacificus
Xiao-Fang LIU1 , Xiang-Yang LI1, 2, Xiao-Yan FU1, 2, Fu-Hou LI2, Wei-Xia WANG2, Kai-Liang LENG1, 3, *
Affiliations
  • 1. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of Sustainable Development of Polar Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao 266071, China
  • 2. School of Ocean Food and Biological Engineering, Jiangsu Ocean University, Jiangsu Key Laboratory of Marine Biotechnology, Lianyungang 222005, China
  • 3. Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao 266237, China
出版时间: 2025-05-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241204004
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目的 分析探究太平洋褶柔鱼(Todarodes pacificus)内脏蛋白肽的抗氧化活性及稳定性。方法 测定太平洋褶柔鱼内脏蛋白肽的自由基清除能力, 评价其抗氧化活性, 并考察温度、酸碱、胃肠道消化等因素对其稳定性的影响。结果 太平洋褶柔鱼内脏蛋白肽具有良好的自由基清除能力, 其清除1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl radical, DPPH)自由基、2,2-联氮-2(3-乙基-苯并噻唑-6-磺酸)二铵盐[2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, ABTS]阳离子自由基、羟自由基、超氧阴离子自由基的半数抑制浓度(half maximal inhibition concentration, IC50)分别为1.38、15.26、0.90、2.21 mg/mL。该蛋白肽的抗氧化活性在25~100 ℃温度范围内较为稳定; 在pH 7.0~8.0条件下其抗氧化活性变化不显著(P>0.05), 但在强酸或强碱条件下其抗氧化活性显著下降(P<0.05); 经体外模拟胃肠道消化后, 该蛋白肽依然能够维持良好的抗氧化活性。结论 太平洋褶柔鱼内脏蛋白肽具备良好的抗氧化活性与稳定性, 但在其加工与贮藏过程中还应注意避免高温与极端酸碱条件的影响。研究成果可为太平洋褶柔鱼内脏资源的高值化利用开辟新途径, 也可为天然抗氧化肽的开发提供科学指导。

太平洋褶柔鱼内脏  /  蛋白肽  /  抗氧化活性  /  稳定性

Objective To explore the antioxidant activity and stability of peptides from the visceral of Todarodes pacificus. Methods Free radical scavenging capacities of peptides from the visceral of Todarodes pacificus were determined, while the effects of temperature, pH and gastrointestinal digestion on the antioxidant activity of peptides from the visceral of Todarodes pacificus were also evaluated. Results The peptides from the visceral of Todarodes pacificus exhibited excellent free radical scavenging abilities, with half maximal inhibition concentration (IC50) values of 1.38, 15.26, 0.90 and 2.21 mg/mL for scavenging 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), 2,2’-azino-bis(3- ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) cationic radicals, hydroxyl radicals, and superoxide anion radicals, respectively. The antioxidant activity of the peptides remained relatively stable within a temperature range of 25-100 °C. Under pH conditions of 7.0-8.0, their antioxidant activity showed no significant change (P>0.05), but it significantly decreased under strongly acidic or alkaline conditions (P<0.05). After in vitro simulated gastrointestinal digestion, the peptides still maintained strong antioxidant activity. Conclusion The peptides from the visceral of Todarodes pacificus has good antioxidant activity and stability, but it shall be noted that the processing and storage of its related products avoid the influence of high temperature and extreme acid-base conditions. The present research will open up new ways for the high-value utilization of the visceral of Todarodes pacificus resources and provide scientific guidance for the innovative development of natural antioxidant peptides.

visceral of Todarodes pacificus  /  peptides  /  antioxidant activity  /  stability
刘小芳, 李向阳, 付晓艳, 李福后, 王伟霞, 冷凯良. 太平洋褶柔鱼内脏蛋白肽的抗氧化活性及稳定性研究. 食品安全质量检测学报, 2025 , 16 (10) : 302 -308 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241204004
Xiao-Fang LIU, Xiang-Yang LI, Xiao-Yan FU, Fu-Hou LI, Wei-Xia WANG, Kai-Liang LENG. Study on the antioxidant activity and stability of peptides from the visceral of Todarodes pacificus[J]. Journal of Food Safety & Quality, 2025 , 16 (10) : 302 -308 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241204004
我国是全球最重要的鱿鱼生产、加工和消费国家之一[1], 据《2023年全国渔业经济统计公报》数据显示, 2023年我国头足类捕捞产量59.36万t, 在远洋渔业捕捞总产量中占比达到25.56%[2]。近年来, 我国鱿鱼年加工量保持在40~50万t, 行业产值超百亿元。鱿鱼产业的发展质量影响着我国远洋捕捞业和水产品加工业的提质升级, 然而目前捕获的鱿鱼主要用来加工生产鱿鱼丝、鱿鱼干、鱿鱼脆片等初级加工产品, 精深加工利用程度较低[1,3]。此外, 在鱿鱼初级加工产品生产过程中所产生的大量副产物, 包括内脏团、皮、眼等, 约达到鱿鱼总重量的25%左右, 而该部分资源通常只被简单加工为鱿鱼膏、鱿鱼油等产品或直接应用于饲料行业[1,3-6], 尚未被最有效加工利用, 造成资源浪费和环境污染等问题, 一定程度上影响着产业高质量发展。太平洋褶柔鱼(Todarodes pacificus, T. pacificus), 又称太平洋斯氏柔鱼、日本飞鱿鱼, 主要分布于西北太平洋海域, 资源量丰富, 是我国主要的鱿鱼捕捞品种[7], 在其加工过程中所产生的内脏团等副产物亦尚未被合理利用。本团队前期研究发现[1], 该内脏团富含蛋白质且氨基酸组成符合联合国粮食及农业组织(Food and Agriculture Organization of the United Nations, FAO)/世界卫生组织(World Health Organization, WHO)规定的优质蛋白标准, 具有良好的开发价值。深入探究太平洋褶柔鱼内脏蛋白肽的功能活性, 挖掘其在健康食品、医药制品等领域的应用价值, 是实现太平洋褶柔鱼资源高值全利用的有效切入点, 并将有助于推动鱿鱼产业实现绿色低碳高质量发展。
近年来, 以水产品及其加工副产物为原料提取制备功能活性肽已成为产业关注的焦点, 其中抗氧化肽是被研究探讨最多的一种活性肽[8]。与合成类抗氧化剂相比, 天然抗氧化肽的功能活性更强、吸收效果更好、安全性更高, 并且加工过程更具环境友好性, 已被广泛应用于健康食品、医药制品等多个领域[9]。目前, 以鱿鱼及其加工副产物为原料制备蛋白肽的工艺研究已初见报道[10-11], 但鲜少有研究对此类蛋白肽的抗氧化活性及稳定性进行较为系统地分析评价。
鉴于此, 本研究以采用太平洋褶柔鱼内脏团为原料酶解制备的蛋白肽为研究对象, 通过测定4种自由基清除能力系统评价其抗氧化活性, 并考察温度、酸碱、胃肠道消化等因素对其稳定性的影响, 以期为太平洋褶柔鱼内脏蛋白肽的精准利用提供理论支持, 为推进实现鱿鱼资源综合高值全利用提供科学指导。
太平洋褶柔鱼内脏团(包含肝脏、胃、生殖腺、消化腺、墨囊等内脏器官): 威海博宇食品有限公司。
碱性蛋白酶Alcalase 2.4 L [2×105 U/mL, 丹麦诺维信(中国)生物技术有限公司]; 胃蛋白酶(250 U/mg)、胰蛋白酶(250 U/mg)、Lowry法蛋白浓度测定试剂盒(PC0030)(北京索莱宝科技有限公司); 1,1-二苯基-2-三硝基苯肼[1,1-diphenyl-2-picrylhydrazyl radical, DPPH, 纯度≥97%, 梯希爱(上海)化成工业发展有限公司]; 2,2-联氮-2(3-乙基-苯并噻唑-6-磺酸)二铵盐[2,2’-azino-bis (3-ethylbenzothiazoline- 6-sulfonic acid) diammonium salt, ABTS, 纯度≥98%, 上海麦克林生化科技有限公司]; 过氧化氢、盐酸(分析纯, 西陇科学股份有限公司); 过硫酸钾、硫酸亚铁、水杨酸、三羟甲基氨基甲烷[Tris (hydroxymethyl) aminomethane, Tris]、邻苯三酚、乙醇、氢氧化钠、粉末活性炭(分析纯, 国药集团化学试剂有限公司)。
BSA224S-CW型电子分析天平(精度0.0001 g, 赛多利斯科学仪器有限公司); ST3100型pH计(常州奥豪斯仪器有限公司); Neofuge 15R型离心机(上海力申科学仪器有限公司); AFA-18-200型膜元件(山东博纳生物科技集团有限公司); XS-Y-MINI-2型有机膜设备(南京诺润机械科技有限公司); CTFD-10P型真空冷冻干燥机(青岛永合创信电子科技有限公司); SHA-B型恒温水浴摇床、HH-4型恒温水浴锅(常州智博瑞仪器制造有限公司); UV1-102II型紫外/可见分光光度计(上海天美科学仪器有限公司)。
参照文献[10]的方法略作修改, 制备太平洋褶柔鱼内脏蛋白肽: 太平洋褶柔鱼内脏团经真空冷冻干燥(冷阱温度-60 ℃, 真空度0.098 Mpa)后, 磨碎, 混合均匀, 按照料液比1:6 (g:mL)加入50 mmol/L pH 7.5磷酸盐缓冲液, 按照质量分数0.35%加入碱性蛋白酶Alcalase 2.4 L, 充分混匀, 于55 ℃反应4 h后, 煮沸灭酶10 min, 静置冷却至50 ℃, 加入10 g/L粉末活性炭, 充分混匀, 吸附脱色1 h后, 于25 ℃、5000 r/min离心15 min, 收集上清液, 采用200 Da截留分子量的滤膜进行脱盐处理, 收集截留液, 经真空冷冻干燥(冷阱温度-60 ℃, 真空度0.098 Mpa)得到太平洋褶柔鱼内脏蛋白肽。按照GB 5009.5—2016《食品安全国家标准 食品中蛋白质的测定》规定的凯氏定氮法测得其总蛋白质含量90.45%; 按照GB 31645—2018《食品安全国家标准 胶原蛋白肽》中附录A规定的方法测得其相对分子质量集中分布于189~6512 Da, 其中3000 Da以下的蛋白肽占比为98.65%。
1) DPPH自由基清除能力测定
取质量浓度为0.2、0.4、0.8、1.6、3.2 mg/mL的太平洋褶柔鱼内脏蛋白肽溶液, 按照文献[9,12]的方法测定: 将1.5 mL不同质量浓度的太平洋褶柔鱼内脏蛋白肽溶液与1.5 mL 0.2 mmol/L DPPH乙醇溶液混合, 室温避光放置, 反应30 min后于517 nm处测定吸光值(A样品)。采用乙醇溶液代替DPPH乙醇溶液测定空白吸光值(A空白), 采用蒸馏水代替太平洋褶柔鱼内脏蛋白肽溶液测定对照吸光值(A对照)。DPPH自由基清除率计算如公式(1):
$\text { DPPH 自由基清除率 } /\%=\left(1-\frac{A_{\text {样品 }}-A_{\text {空白 }}}{A_{\text {对照 }}}\right) \times 100 \%$
2) ABTS阳离子自由基清除能力测定
取质量浓度为1.6、3.2、6.4、12.8、25.6 mg/mL的太平洋褶柔鱼内脏蛋白肽溶液, 按照文献[9,12]的方法测定: 将25 mL 7 mmol/L ABTS乙醇溶液与0.44 mL 2.4 mmol/L过硫酸钾溶液混合, 室温避光贮存12~16 h, 采用乙醇稀释至734 nm处吸光度值为0.70±0.02, 配制成ABTS试剂。将0.15 mL不同质量浓度的太平洋褶柔鱼内脏蛋白肽溶液与3 mL ABTS试剂混合, 室温避光放置, 反应20 min后于734 nm处测定吸光值(B样品)。采用ABTS乙醇溶液代替ABTS试剂测定空白吸光值(B空白), 采用蒸馏水代替太平洋褶柔鱼内脏蛋白肽溶液测定对照吸光值(B对照)。ABTS阳离子自由基清除率计算如公式(2):
$\text { ABTS 阳离子自由基清除率 } /\%=\left(1-\frac{B_{\text {样品 }}-B_{\text {空白 }}}{B_{\text {对照 }}}\right) \times 100 \%$
3)羟自由基清除能力测定
取质量浓度为0.125、0.250、0.500、1.000、2.000 mg/mL的太平洋褶柔鱼内脏蛋白肽溶液, 按照文献[9,12]的方法测定: 将1 mL不同质量浓度的太平洋褶柔鱼内脏蛋白肽溶液与0.5 mL 9 mmol/L硫酸亚铁溶液、1 mL 9 mmol/L水杨酸乙醇溶液、1 mL 4.4 mmol/L过氧化氢溶液和2 mL蒸馏水混合, 置于37 ℃水浴, 反应30 min后于510 nm下测定吸光值(C样品)。采用乙醇代替水杨酸乙醇溶液测定空白吸光值(C空白), 采用蒸馏水代替太平洋褶柔鱼内脏蛋白肽溶液测定对照吸光值(C对照)。羟自由基清除率计算如公式(3):
$\text { 羟自由基清除率 } /\%=\left(1-\frac{C_{\text {样品 }}-C_{\text {空白 }}}{C_{\text {对照 }}}\right) \times 100 \%$
4)超氧阴离子自由基清除能力测定
取质量浓度为0.4、0.8、1.6、3.2、6.4 mg/mL的太平洋褶柔鱼内脏蛋白肽溶液, 按照文献[9,12]的方法测定: 将0.1 mL不同质量浓度的太平洋褶柔鱼内脏蛋白肽溶液与4.5 mL 50 mmol/L Tris-盐酸缓冲液(pH 8.2)充分混合, 在25 ℃水浴保温25 min, 然后立刻加入0.1 mL 3 mmol/L邻苯三酚溶液(使用10 mmol/L盐酸溶液配制), 迅速混匀后于25 ℃水浴反应5 min, 结束后立即在325 nm下测定吸光值(D样品)。采用蒸馏水代替邻苯三酚溶液测定空白吸光值(D空白), 采用蒸馏水代替太平洋褶柔鱼内脏蛋白肽溶液测定对照吸光值(D对照)。超氧阴离子自由基清除率计算如公式(4):
$\text { 超氧阴离子自由基清除率 } /\%=\left(1-\frac{D_{\text {样品 }}-D_{\text {空白 }}}{D_{\text {对照 }}}\right) \times 100 \%$
1)温度对太平洋褶柔鱼内脏蛋白肽抗氧化活性的影响
按照文献[9,12-14]的方法, 取质量浓度为0.9 mg/mL太平洋褶柔鱼内脏蛋白肽溶液, 测定其于25、37、60、80、100 ℃水浴保温1 h后的羟自由基清除率。
2) pH对太平洋褶柔鱼内脏蛋白肽抗氧化活性的影响
按照文献[9,14-15]的方法, 取质量浓度为0.9 mg/mL太平洋褶柔鱼内脏蛋白肽溶液, 测定其处于pH 2.0、4.0、6.0、7.0、8.0、10.0、12.0条件下1 h后的羟自由基清除率。
3)体外模拟胃肠道消化对太平洋褶柔鱼内脏蛋白肽抗氧化活性的影响
按照文献[9,12]方法, 取质量浓度为0.9 mg/mL太平洋褶柔鱼内脏蛋白肽溶液, 调节pH至2.0, 加入底物质量分数4.0%的胃蛋白酶, 混合均匀, 于37 ℃反应1.5 h, 而后将消化液分为两等份, 一份煮沸10 min终止反应, 即得胃消化样品; 另一份继续反应, 调节pH至7.5, 加入底物质量分数4.0%的胰蛋白酶, 混合均匀, 于37 ℃反应2 h, 而后煮沸10 min终止反应, 即得胃肠消化样品; 分别测定消化前、胃消化、胃肠消化样品的羟自由基清除率。
实验平行测定3次, 实验数据采用“平均值±标准偏差”的形式表示, 采用IBM SPSS 13.0、Excel 2019、Origin 2022等软件进行数据处理和图形绘制。采用单因素方差分析进行组间多重比较, 以P<0.05为差异具有统计学意义。
太平洋褶柔鱼内脏蛋白肽的DPPH自由基清除能力测定结果见图1。由图1可知, 太平洋褶柔鱼内脏蛋白肽质量浓度在0.2~3.2 mg/mL范围时, DPPH自由基清除率为(1.96±0.89)%~(72.21±3.32)%; 随着蛋白肽质量浓度的升高, 其DPPH自由基清除率显著增加(P<0.05)。经计算, 太平洋褶柔鱼内脏蛋白肽清除DPPH自由基的半数抑制浓度(half maximal inhibition concentration, IC50)为1.38 mg/mL。与已有报道的鲍鱼内脏蛋白肽[16](IC50值1.51 mg/mL)、鲣鱼加工副产物蛋白肽[17](IC50值10.02 mg/mL)、马面鱼皮胶原蛋白肽[18](IC50值1.80 mg/mL)等水产品加工副产物来源蛋白肽相比, 太平洋褶柔鱼内脏蛋白肽具有更强的DPPH自由基清除能力。
太平洋褶柔鱼内脏蛋白肽的ABTS阳离子自由基清除能力测定结果见图2。由图2可知, 太平洋褶柔鱼内脏蛋白肽质量浓度在1.6~25.6 mg/mL范围时, ABTS阳离子自由基清除率为(4.17±1.31)%~(76.52±0.01)%; 随着蛋白肽质量浓度的升高, 其ABTS阳离子自由基清除率整体上显著增加(P<0.05)。经计算, 太平洋褶柔鱼内脏蛋白肽清除ABTS阳离子自由基的IC50值为15.26 mg/mL。太平洋褶柔鱼内脏蛋白肽的ABTS阳离子自由基清除能力优于鲍鱼内脏蛋白肽[19](IC50值17.41 mg/mL), 弱于西班牙鲭鱼皮胶原蛋白肽[20](IC50值1.07 mg/mL)。
太平洋褶柔鱼内脏蛋白肽的羟自由基清除能力测定结果见图3。由图3可知, 太平洋褶柔鱼内脏蛋白肽质量浓度在0.125~2.000 mg/mL范围时, 羟自由基清除率为(7.53±2.59)%~(91.16±0.99)%; 随着蛋白肽质量浓度的升高, 其羟自由基清除率整体上显著增加(P<0.05)。经计算, 太平洋褶柔鱼内脏蛋白肽清除羟自由基的IC50值为0.90 mg/mL。与已有报道的北太平洋鱿鱼内脏蛋白肽[21](IC50值1.85 mg/mL)、鲣鱼加工副产物蛋白肽[17](IC50值2.94 mg/mL)、鳐鱼软骨蛋白肽[22](IC50值5.03 mg/mL)等水产品加工副产物来源蛋白肽相比, 太平洋褶柔鱼内脏蛋白肽具有更强的羟自由基清除能力。与其清除DPPH自由基(IC50值1.38 mg/mL)、ABTS阳离子自由基(IC50值15.26 mg/mL)和超氧阴离子自由基(IC50值2.21 mg/mL)的能力相比, 太平洋褶柔鱼内脏蛋白肽清除羟自由基(IC50值0.90 mg/mL)的能力更强, 反应更为灵敏。因此后续采用羟自由基清除率作为评价指标考察太平洋褶柔鱼内脏蛋白肽在不同温度、酸碱及模拟胃肠道消化条件下的稳定性。
太平洋褶柔鱼内脏蛋白肽的超氧阴离子自由基清除能力测定结果见图4。由图4可知, 太平洋褶柔鱼内脏蛋白肽质量浓度在0.4~6.4 mg/mL时, 羟自由基清除率为(33.63±2.92)%~(57.46±3.08)%; 随着蛋白肽质量浓度的升高, 其超氧阴离子自由基清除率整体上显著增加(P<0.05)。经计算, 太平洋褶柔鱼内脏蛋白肽清除超氧阴离子自由基的IC50值为2.21 mg/mL。与已有报道的皱纹盘鲍内脏蛋白肽[23](IC50值4.92 mg/mL)、大青鲨鱼皮胶原蛋白肽[24](IC50值7.32 mg/mL)、鲽鱼鱼皮胶原蛋白肽[25](IC50值7.98 mg/mL)等水产品加工副产物来源蛋白肽相比, 太平洋褶柔鱼内脏蛋白肽具有更强的超氧阴离子自由基清除能力。
图5可知, 太平洋褶柔鱼内脏蛋白肽在25~100 ℃范围内, 其羟自由基清除率呈现先升高后下降的变化趋势; 在25~60 ℃范围内, 其羟自由基清除率随着温度升高而略有提高, 但变化不显著(P>0.05); 在温度超过60 ℃后, 随着温度的继续升高, 其羟自由基清除率出现下降(P<0.05); 在温度到达100 ℃时, 其羟自由基清除率仍能保持原有的86.87%; 这与ZHU等[26]的研究结果较为一致。适当升温能使多肽的一级结构发生改变, 生成更多的短链多肽, 暴露出更多的活性基团, 从而提高其抗氧化活性; 但随着温度继续升高, 多肽结构中的化学键遭到破坏, 其抗氧化活性出现下降[27]。综上, 太平洋褶柔鱼内脏蛋白肽拥有良好的热稳定性, 但在其相关产品的加工过程中仍需要注意避免高温的影响。
图6可知, 在pH为2.0~12.0范围内, 随着pH的增加, 太平洋褶柔鱼内脏蛋白肽的羟自由基清除率呈现先升高后降低的变化趋势, 且整体变化显著(P<0.05); pH为7.0时, 太平洋褶柔鱼内脏蛋白肽的羟自由基清除率最高; 酸性条件对其抗氧化活性的影响大于碱性条件(P<0.05), 在pH为2.0和12.0时, 其羟自由基清除率分别降至原有的79.19%和86.41%。在酸性条件下, 多肽中的酰胺键易被催化水解断裂, 而在碱性条件下, 多肽易发生外消旋反应, 部分L-型氨基酸转变成D-型氨基酸, 肽链构象的改变抑制了其与自由基结合的能力, 最终造成多肽的抗氧化活性降低[28-29]。综上, 在太平洋褶柔鱼内脏蛋白肽相关产品的加工过程中要注意避免强酸强碱的影响。
图7可知, 太平洋褶柔鱼内脏蛋白肽在模拟胃肠消化过程中, 其羟自由基清除率呈现先降低后升高的变化趋势; 太平洋褶柔鱼内脏蛋白肽经模拟胃消化处理后, 其羟自由基清除率由(52.54±1.24)%下降至(51.37±1.74)%, 变化不显著(P>0.05); 再经模拟肠消化处理后, 其羟自由基清除率有所提高, 升高至(59.64±1.51)%, 变化显著(P<0.05); 这与KETNAWA等[30]的研究结果较为一致。多肽的功能活性依赖于其独特结构, 太平洋褶柔鱼内脏蛋白肽在模拟胃消化过程中, 部分活性多肽受到破坏, 导致其与羟自由基发生反应的能力减弱; 而在模拟肠消化过程中, 长链多肽被进一步降解为短链多肽, 暴露出更多的活性基团, 使其与羟自由基发生反应的能力有所增强。综上, 太平洋褶柔鱼内脏蛋白肽经模拟胃肠道消化后仍能保持良好的抗氧化活性, 较好的消化稳定性有助于其在生物体内发挥有效的抗氧化作用。
本研究对太平洋褶柔鱼内脏蛋白肽抗氧化活性及稳定性进行了分析与评价。抗氧化活性研究结果表明, 太平洋褶柔鱼内脏蛋白肽具有良好的自由基清除能力, 其清除DPPH自由基、ABTS阳离子自由基、羟自由基、超氧阴离子自由基的IC50值分别为1.38、15.26、0.90、2.21 mg/mL。稳定性研究结果表明, 太平洋褶柔鱼内脏蛋白肽具有良好的热稳定性和消化稳定性; pH对其稳定性具有一定影响, 在强酸、强碱条件下抗氧化活性明显降低。未来研究可对太平洋褶柔鱼内脏蛋白肽的制备工艺进行优化, 对抗氧化活性最优组分进行分离纯化和筛选鉴定, 并深入解析其作用机制, 定向开发健康食品、化妆品、医药制品等领域的功能产品。本研究对于推进鱿鱼加工副产物资源合理利用和新型抗氧化肽高值产品开发具有积极意义。
  • 青岛市关键技术攻关及产业化示范类项目(23-1-3-hysf-1-hy)
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2025年第16卷第10期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241204004
  • 接收时间:2024-12-04
  • 首发时间:2025-07-15
  • 出版时间:2025-05-25
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  • 收稿日期:2024-12-04
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青岛市关键技术攻关及产业化示范类项目(23-1-3-hysf-1-hy)
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    1.中国水产科学研究院黄海水产研究所, 农业农村部极地渔业可持续利用重点实验室, 青岛 266071
    2.江苏海洋大学海洋食品与生物工程学院, 江苏省海洋生物资源与环境重点实验室, 连云港 222005
    3.青岛海洋科技中心, 海洋药物与生物制品功能实验室, 青岛 266237

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* 冷凯良(1966—), 男, 研究员, 主要研究方向为水产品加工与利用。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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