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Article(id=1216517521973367026, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1216517514570417012, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250121005, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1737388800000, receivedDateStr=2025-01-21, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1767969979041, onlineDateStr=2026-01-09, pubDate=1755187200000, pubDateStr=2025-08-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1767969979041, onlineIssueDateStr=2026-01-09, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1767969979041, creator=13701087609, updateTime=1767969979041, updator=13701087609, issue=Issue{id=1216517514570417012, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='15', pageStart='1', pageEnd='322', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1767969977276, creator=13701087609, updateTime=1768211590858, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1217530915467743720, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1216517514570417012, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1217530915467743721, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1216517514570417012, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=213, endPage=221, ext={EN=ArticleExt(id=1216517523156160810, articleId=1216517521973367026, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Preparation and physicochemical properties research of Juglans regia L. polypeptide-selenium chelate, columnId=1151923894560060071, journalTitle=Journal of Food Safety & Quality, columnName=Food Nutrition and Functional Foods, runingTitle=null, highlight=null, articleAbstract=

Objective To study the preparation and physicochemical properties of polypeptide-selenium chelate from Juglans regia L.. Methods Juglans regia L. protein was extracted by alkali-soluble acid precipitation method, and Juglans regia L. polypeptide was prepared by alkaline protease. Juglans regia L. polypeptide reacted with sodium selenite to produce Juglans regia L. polypeptide-selenium chelate. The preparation technology was optimized by single factor test and orthogonal test. Results When the mass ratio of polypeptide to sodium selenite was 3:1, the chelating temperature was 45 ℃, the chelating time was 50 min, and the pH was 8, the chelating rate was 58.99%±1.44%. The chelate was resistant to heat, acid and alkali, had good digestive stability and high bioavailability of selenium. Compared with Juglans regia L. polypeptide, Juglans regia L. polypeptide-selenium chelate had stronger scavenging ability of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical, hydroxyl free radical and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cation free radical. Conclusion In this study, Juglans regia L. polypeptide-selenium chelate is successfully prepared through optimized synthesis conditions. The resulting complex demonstrates favorable physicochemical properties and significant antioxidant capacity, providing a potential raw material reference for developing selenium-enriched nutritional supplements.

, correspAuthors=Jing WANG, 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=Jing WANG, Dan FANG, Shi-Jia LIU, Ya-Zhi DENG, Zhao-Yu MAO, Jia LI, Jing-Yang WANG, Li-Ping FENG), CN=ArticleExt(id=1216517530563302024, articleId=1216517521973367026, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=核桃多肽-硒螯合物的制备及其理化性质研究, 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 探究核桃多肽-硒螯合物的制备方法及其理化性质。方法 采用碱溶酸沉法提取核桃蛋白, 经碱性蛋白酶酶解后制得核桃多肽。核桃多肽再与亚硒酸钠反应, 生成核桃多肽-硒螯合物。通过单因素试验和正交试验, 优化制备工艺。结果 在多肽与亚硒酸钠质量比为3:1、螯合温度45 ℃、螯合时间50 min、pH 8的条件下, 硒螯合率达到最高, 为58.99%±1.44%。该螯合物耐热、耐酸碱, 消化稳定性好, 硒生物利用率高。与核桃多肽相比, 核桃多肽-硒螯合物具有更强的1,1-二苯基-2-苦味酰肼(1,1-diphenyl-2-picrylhydrazyl, DPPH)自由基、羟基自由基和2,2'-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐[2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), ABTS]阳离子自由基清除能力。结论 本研究制备了核桃多肽-硒螯合物, 优化了制备工艺, 并发现其具有良好的理化性质和抗氧化能力, 为开发补硒产品提供了原料参考。

, correspAuthors=王静, authorNote=null, correspAuthorsNote=
*王静(1985—), 女, 博士, 副教授, 主要研究方向为食品功能因子的研究与开发。E-mail:
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注: 图中不同小写字母表示差异显著(P<0.05)。

, figureFileSmall=KXVrwxsu0ITE/qS3lC7UCg==, figureFileBig=N73xrNjjyTKT0DysvdSPtQ==, tableContent=null), ArticleFig(id=1217127913820963722, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Fig.2, caption=Ultraviolet spectra of Juglans regia L. polypeptide and Juglans regia L. polypeptide-selenium chelates, figureFileSmall=DAGT0s8jfomkVtG/HvsuMA==, figureFileBig=WSTX4qyudp9YLsBMGy43UQ==, tableContent=null), ArticleFig(id=1217127913934209936, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=图2, caption=核桃多肽与核桃多肽-硒螯合物的紫外光谱图, figureFileSmall=DAGT0s8jfomkVtG/HvsuMA==, figureFileBig=WSTX4qyudp9YLsBMGy43UQ==, tableContent=null), ArticleFig(id=1217127914093593501, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Fig.3, caption=Fluorescence spectra of Juglans regia L. polypeptide and Juglans regia L. polypeptide-selenium chelates, figureFileSmall=ca2bMo3/hZGdyeItaX/W2g==, figureFileBig=+h3kVT99zPzMBIt35NLRbg==, tableContent=null), ArticleFig(id=1217127914190062499, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=图3, caption=核桃多肽与核桃多肽-硒螯合物的荧光光谱图, figureFileSmall=ca2bMo3/hZGdyeItaX/W2g==, figureFileBig=+h3kVT99zPzMBIt35NLRbg==, tableContent=null), ArticleFig(id=1217127914282337195, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Fig.4, caption=Solubility of Juglans regia L. polypeptide-selenium chelates at different pH, figureFileSmall=vT9jXqndCCk09eGEy40NpA==, figureFileBig=VWU3S3oiVEqlxNxZj75+Ug==, tableContent=null), ArticleFig(id=1217127914395583409, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=图4, caption=核桃多肽-硒螯合物在不同pH下的溶解度, figureFileSmall=vT9jXqndCCk09eGEy40NpA==, figureFileBig=VWU3S3oiVEqlxNxZj75+Ug==, tableContent=null), ArticleFig(id=1217127914504635319, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Fig.5, caption=Relative proportions of selenium content of Juglans regia L. polypeptide-selenium chelates at different temperatures, figureFileSmall=4agMmt5RMWkM3dQDPRw+eQ==, figureFileBig=CAKyc/GDJPZfRVYhTid7Ug==, tableContent=null), ArticleFig(id=1217127915788092349, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=图5, caption=核桃多肽-硒螯合物在不同温度下的硒含量相对比例, figureFileSmall=4agMmt5RMWkM3dQDPRw+eQ==, figureFileBig=CAKyc/GDJPZfRVYhTid7Ug==, tableContent=null), ArticleFig(id=1217127915922310083, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Fig.6, caption=Relative proportions of selenium content of Juglans regia L. polypeptide-selenium chelates at different pH, figureFileSmall=AfDPHYJM793yIxitE780bw==, figureFileBig=vIg9ugvtlpyznRIlM4ci7A==, tableContent=null), ArticleFig(id=1217127916039750604, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=图6, caption=核桃多肽-硒螯合物在不同pH下的硒含量相对比例, figureFileSmall=AfDPHYJM793yIxitE780bw==, figureFileBig=vIg9ugvtlpyznRIlM4ci7A==, tableContent=null), ArticleFig(id=1217127916148802514, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Fig.7, caption=Relative proportions of selenium content of Juglans regia L. polypeptide-selenium chelates under different digestion modes, figureFileSmall=2QuyI8PVghdnNFhaJla6dw==, figureFileBig=Uja+iiqFUWOjTYPP1eS8GQ==, tableContent=null), ArticleFig(id=1217127916232688596, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=图7, caption=核桃多肽-硒螯合物在不同消化方式下的硒含量相对比例, figureFileSmall=2QuyI8PVghdnNFhaJla6dw==, figureFileBig=Uja+iiqFUWOjTYPP1eS8GQ==, tableContent=null), ArticleFig(id=1217127916354323424, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Fig.8, caption=DPPH radical scavenging capacity of Juglans regia L. polypeptide and Juglans regia L. polypeptide-selenium chelates, figureFileSmall=Io5p4yJgogJCNfs8PO6VjQ==, figureFileBig=5dvyqybejTpnY5pcInFi5Q==, tableContent=null), ArticleFig(id=1217127916484346851, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=图8, caption=核桃多肽与核桃多肽-硒螯合物的DPPH自由基清除能力, figureFileSmall=Io5p4yJgogJCNfs8PO6VjQ==, figureFileBig=5dvyqybejTpnY5pcInFi5Q==, tableContent=null), ArticleFig(id=1217127916610175981, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Fig.9, caption=Hydroxyl radical scavenging capacity of Juglans regia L. polypeptide and Juglans regia L. polypeptide-selenium chelates, figureFileSmall=pHAQd9dPj0XCVkNMhUne4g==, figureFileBig=F9313T6cAadry/ev/D6HmA==, tableContent=null), ArticleFig(id=1217127916756976631, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=图9, caption=核桃多肽与核桃多肽-硒螯合物的羟基自由基清除能力, figureFileSmall=pHAQd9dPj0XCVkNMhUne4g==, figureFileBig=F9313T6cAadry/ev/D6HmA==, tableContent=null), ArticleFig(id=1217127916866028542, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Fig.10, caption=ABTS cation radical scavenging capacity of Juglans regia L. polypeptide and Juglans regia L. polypeptide-selenium chelates, figureFileSmall=XfHEyjH9xSW1ez2IOKpHYQ==, figureFileBig=kxJz2gJLZ/o5mqcH1RrAgQ==, tableContent=null), ArticleFig(id=1217127916962496515, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=图10, caption=核桃多肽与核桃多肽-硒螯合物的ABTS阳离子自由基清除能力, figureFileSmall=XfHEyjH9xSW1ez2IOKpHYQ==, figureFileBig=kxJz2gJLZ/o5mqcH1RrAgQ==, tableContent=null), ArticleFig(id=1217127917071548425, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Table 1, caption=

Level table of single-factor research

, figureFileSmall=null, figureFileBig=null, tableContent=
影响因素 因素水平
多肽与亚硒酸钠质量比 1:2 1:1 2:1 3:1 4:1
螯合温度/℃ 30 40 50 60 70
螯合时间/min 20 30 40 50 60
pH 6 7 8 9 10
), ArticleFig(id=1217127917188988943, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=表1, caption=

单因素研究水平表

, figureFileSmall=null, figureFileBig=null, tableContent=
影响因素 因素水平
多肽与亚硒酸钠质量比 1:2 1:1 2:1 3:1 4:1
螯合温度/℃ 30 40 50 60 70
螯合时间/min 20 30 40 50 60
pH 6 7 8 9 10
), ArticleFig(id=1217127917314818070, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Table 2, caption=

Factor-level table of orthogonal test

, figureFileSmall=null, figureFileBig=null, tableContent=
水平 螯合物制备的影响因素
A(核桃多肽与
亚硒酸钠质量比)		 B (pH) C(螯合时间)
/min		 D(螯合温度)
/℃
1 5:2 7 45 45
2 3:1 8 50 50
3 7:2 9 55 55
), ArticleFig(id=1217127917516144674, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=表2, caption=

正交试验因素水平表

, figureFileSmall=null, figureFileBig=null, tableContent=
水平 螯合物制备的影响因素
A(核桃多肽与
亚硒酸钠质量比)		 B (pH) C(螯合时间)
/min		 D(螯合温度)
/℃
1 5:2 7 45 45
2 3:1 8 50 50
3 7:2 9 55 55
), ArticleFig(id=1217127917616807980, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=EN, label=Table 3, caption=

Analysis table of orthogonal test results for selenium chelation rate

, figureFileSmall=null, figureFileBig=null, tableContent=
试验号 因素 硒螯
合率/%
A B C D
1 1 1 1 1 27.42
2 1 2 3 2 33.78
3 1 3 2 3 27.90
4 2 1 3 3 30.17
5 2 2 2 1 58.99
6 2 3 1 2 40.34
7 3 1 2 2 39.73
8 3 2 1 3 39.38
9 3 3 3 1 38.34
K1 89.09 97.32 107.14 124.75
K2 129.50 132.14 126.61 113.85
K3 117.45 106.58 102.29 97.44
k1 29.70 32.44 35.71 41.58
k2 43.17 44.05 42.20 37.95
k3 39.15 35.53 34.10 32.48
R 13.47 8.52 8.11 5.47
), ArticleFig(id=1217127917725859889, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517521973367026, language=CN, label=表3, caption=

硒螯合率正交试验结果分析表

, figureFileSmall=null, figureFileBig=null, tableContent=
试验号 因素 硒螯
合率/%
A B C D
1 1 1 1 1 27.42
2 1 2 3 2 33.78
3 1 3 2 3 27.90
4 2 1 3 3 30.17
5 2 2 2 1 58.99
6 2 3 1 2 40.34
7 3 1 2 2 39.73
8 3 2 1 3 39.38
9 3 3 3 1 38.34
K1 89.09 97.32 107.14 124.75
K2 129.50 132.14 126.61 113.85
K3 117.45 106.58 102.29 97.44
k1 29.70 32.44 35.71 41.58
k2 43.17 44.05 42.20 37.95
k3 39.15 35.53 34.10 32.48
R 13.47 8.52 8.11 5.47
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核桃多肽-硒螯合物的制备及其理化性质研究
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王静 * , 方丹 , 刘诗佳 , 邓雅之 , 毛招羽 , 李佳 , 王婧扬 , 冯莉萍
食品安全质量检测学报 | 食品营养及功能性食品 2025,16(15): 213-221
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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(15): 213-221
核桃多肽-硒螯合物的制备及其理化性质研究
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王静* , 方丹, 刘诗佳, 邓雅之, 毛招羽, 李佳, 王婧扬, 冯莉萍
作者信息
  • 陕西学前师范学院生物食品与化学学院, 西安 710100

通讯作者:

*王静(1985—), 女, 博士, 副教授, 主要研究方向为食品功能因子的研究与开发。E-mail:
Preparation and physicochemical properties research of Juglans regia L. polypeptide-selenium chelate
Jing WANG* , Dan FANG, Shi-Jia LIU, Ya-Zhi DENG, Zhao-Yu MAO, Jia LI, Jing-Yang WANG, Li-Ping FENG
Affiliations
  • College of Biology, Food and Chemistry, Shaanxi Xueqian Normal University, Xi'an 710100, China
出版时间: 2025-08-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250121005
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摘要
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目的 探究核桃多肽-硒螯合物的制备方法及其理化性质。方法 采用碱溶酸沉法提取核桃蛋白, 经碱性蛋白酶酶解后制得核桃多肽。核桃多肽再与亚硒酸钠反应, 生成核桃多肽-硒螯合物。通过单因素试验和正交试验, 优化制备工艺。结果 在多肽与亚硒酸钠质量比为3:1、螯合温度45 ℃、螯合时间50 min、pH 8的条件下, 硒螯合率达到最高, 为58.99%±1.44%。该螯合物耐热、耐酸碱, 消化稳定性好, 硒生物利用率高。与核桃多肽相比, 核桃多肽-硒螯合物具有更强的1,1-二苯基-2-苦味酰肼(1,1-diphenyl-2-picrylhydrazyl, DPPH)自由基、羟基自由基和2,2'-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐[2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), ABTS]阳离子自由基清除能力。结论 本研究制备了核桃多肽-硒螯合物, 优化了制备工艺, 并发现其具有良好的理化性质和抗氧化能力, 为开发补硒产品提供了原料参考。

核桃多肽-硒螯合物  /  理化性质  /  稳定性  /  体外消化  /  抗氧化能力

Objective To study the preparation and physicochemical properties of polypeptide-selenium chelate from Juglans regia L.. Methods Juglans regia L. protein was extracted by alkali-soluble acid precipitation method, and Juglans regia L. polypeptide was prepared by alkaline protease. Juglans regia L. polypeptide reacted with sodium selenite to produce Juglans regia L. polypeptide-selenium chelate. The preparation technology was optimized by single factor test and orthogonal test. Results When the mass ratio of polypeptide to sodium selenite was 3:1, the chelating temperature was 45 ℃, the chelating time was 50 min, and the pH was 8, the chelating rate was 58.99%±1.44%. The chelate was resistant to heat, acid and alkali, had good digestive stability and high bioavailability of selenium. Compared with Juglans regia L. polypeptide, Juglans regia L. polypeptide-selenium chelate had stronger scavenging ability of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical, hydroxyl free radical and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cation free radical. Conclusion In this study, Juglans regia L. polypeptide-selenium chelate is successfully prepared through optimized synthesis conditions. The resulting complex demonstrates favorable physicochemical properties and significant antioxidant capacity, providing a potential raw material reference for developing selenium-enriched nutritional supplements.

Juglans regia L. polypeptide-selenium chelate  /  physicochemical properties  /  stability  /  in vitro digestion  /  antioxidant capacity
王静, 方丹, 刘诗佳, 邓雅之, 毛招羽, 李佳, 王婧扬, 冯莉萍. 核桃多肽-硒螯合物的制备及其理化性质研究. 食品安全质量检测学报, 2025 , 16 (15) : 213 -221 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250121005
Jing WANG, Dan FANG, Shi-Jia LIU, Ya-Zhi DENG, Zhao-Yu MAO, Jia LI, Jing-Yang WANG, Li-Ping FENG. Preparation and physicochemical properties research of Juglans regia L. polypeptide-selenium chelate[J]. Journal of Food Safety & Quality, 2025 , 16 (15) : 213 -221 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250121005
正文
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0 引言
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硒, 作为一种至关重要的微量元素, 对人类健康、生长发育至关重要。缺乏硒元素会引起克山病、大骨节病等慢性病以及贫血、糖尿病等疾病, 严重影响健康[1]。因人体无法合成硒且食物来源不足, 导致人体普遍缺硒, 成为公共健康问题。有机硒相比无机硒(如亚硒酸钠)展现出高生物利用度与低毒性的优势[2], 成为安全、高效的补硒新选择。
当前, 已有研究表明可以通过多肽与硒盐螯合得到有机硒化合物, 使无机硒转化成硒螯合物。赵立娜等[3]成功将菌草灵芝多肽与亚硒酸钠进行螯合, 并利用响应面法对其制备工艺进行优化, 得出多肽-硒螯合物制备的最佳工艺条件。YE等[4]也制备出了具有抗氧化活性的大豆多肽-硒螯合物。意如等[5]综述了近年来硒盐与多肽螯合成硒化合物的研究进展, 并总结了硒肽在人体健康方面的作用。
核桃粕作为核桃油加工的主要副产物, 具有原料来源广、营养价值高、成本低廉的天然优势, 却长期面临利用率低、附加值缺失的产业化困境。本研究基于资源高值化理念, 系统探究了从核桃粕中制备核桃多肽-硒螯合物的完整工艺体系, 并研究了其结构表征与理化性质。该研究通过开发副产物资源化利用的创新性技术路径, 不仅实现了核桃加工产业链的延伸拓展, 其制备的功能性食品基料更展现出双重应用价值: 在营养强化领域, 通过优化硒元素生物利用度, 为新型补硒制剂的开发提供理论支撑; 在功能食品开发方面, 基于多肽-硒复合体系特有的抗氧化活性, 为膳食干预硒缺乏相关疾病开辟了创新研究方向。研究结果对提升特色农副产品深加工技术水平、推动功能性食品原料创新具有重要实践意义。
1 材料与方法
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1.1 材料与试剂
碱性蛋白酶(alcalase 2.4 L, 酶活力1×105 U/g)、1,1-二苯基-2-苦味酰肼(1,1-diphenyl-2-picrylhydrazyl, DPPH)(分析纯, 合肥博美生物科技有限责任公司); 二水合钼酸钠、乙二胺四乙酸二钠(ethylenediaminetetraacetic acid disodium salt, EDTA-2Na)、氢氧化钠(NaOH)(分析纯, 广东光华科技股份有限公司); 氯化氢标准滴定溶液(分析纯, 山东普惠分化学科技有限公司); 无水乙醇、硫酸亚铁(分析纯, 天津市天力化学试剂有限公司); 亚硒酸钠[分析纯, 西格玛奥德里奇(上海)贸易有限公司]; 浓硫酸、甲苯、高氯酸(分析纯, 天津政成化学制品有限公司); 3,3'-二氨基联苯胺(分析纯, 上海麦克林生化科技有限公司); 过硫酸钾、过氧化氢溶液(分析纯, 天津市大茂化学试剂厂); 胃蛋白酶(酶活力3×103 U/g)、胰蛋白酶(酶活力2.5×102 U/g)(西安仟诺生物工程有限责任公司); 邻苯二甲酸氢钾、混合磷酸盐、四硼酸钠(分析纯, 上海佑科仪器仪表有限公司); 2,2'-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐[2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), ABTS](分析纯, 上海恒远生物科技有限公司); 水杨酸(分析纯, 天津市科密欧化学试剂有限公司)。
1.2 仪器与设备
FA2003型电子天平(精度0.1 mg, 上海菁海仪器有限公司); KQ5200DE型数控超声波清洗器(昆山市超声仪器有限公司); V-T5型可见分光光度计(上海光谱仪器有限公司); DK-98-IIA型电热恒温水浴锅(天津市泰斯特仪器有限公司); PHS-3E 型实验室pH计(上海佑科仪器仪表有限公司); DW-HL34型超低温冰箱(安徽中科都菱商用电器股份有限公司); TDZ5型台式低速离心机(湖南赫西仪器装备有限公司); LGJ-18C型真空冷冻干燥机(北京四环科学仪器厂有限公司); DF-101S型集热式恒温磁力搅拌器(巩义市予华仪有限责任公司); ZYM-II-20T型微量分析型超纯水机(四川卓越水处理设备有限公司); RE-52A型旋转蒸发器(上海亚荣生化仪器厂); SH220N型石墨消解仪(山东海能科学仪器有限公司); U-T1810型紫外可见分光光度计(上海屹谱仪器制造有限公司)。
1.3 试验方法
1.3.1 脱脂核桃粕粉末制备工艺流程
为了减少核桃粕中油脂成分对核桃多肽提取率的影响, 需要进行前处理。脱脂工艺流程参照代晹鑫等[6]的方法, 略作改动。将一定质量的核桃粕放入超微粉碎机中进行粉碎, 过筛后, 按照配比核桃粕溶液, 静置12 h进行脱脂。将脱脂核桃粕液3000 r/min离心20 min后, 取离心管沉淀放入恒温干燥箱中干燥(30 ℃, 12 h), 即可制得脱脂核桃粕粉末。
1.3.2 核桃粕总蛋白制备工艺流程
参照马开创[7]的方法, 略作改动。将一定质量的脱脂核桃粕粉末按照比例配制成混合液, 用1 mol/L的NaOH溶液调节混合液pH为8.5, 再进行超声浸提, 浸提完成后, 将核桃粕溶液进行离心, 除去油脂层和下层沉淀后, 用1 mol/L的HCl溶液调节pH为4.5, 将溶液再次离心, 取下层沉淀, 弃去上清液。用超纯水将蛋白质沉淀水洗至中性后倒入培养皿中, 放入真空冷冻干燥机中干燥, 干燥24 h后即可制得核桃粕总蛋白粉, 在-20 ℃下保存, 备用。
1.3.3 核桃蛋白肽制备工艺流程
称取一定质量的核桃粕总蛋白粉, 按核桃粕总蛋白:水=1:25 (W:W)的比例加入蒸馏水, 配制成底物浓度为4%的核桃粕蛋白质溶液, 将pH调至8.8, 根据蛋白粉质量计算出所需添加碱性蛋白酶的质量, 在恒温磁力搅拌器(55 ℃)中酶解2 h。然后100 ℃下灭酶10 min, 冷却至室温, 在转速4000 r/min下离心25 min, 取上清液进行真空冷冻干燥, 即制得核桃多肽粉, 保存于-20 ℃下, 备用[8]。
核桃多肽得率可用式(1)计算:
核桃多肽得率/%=(mc/mb)×100%
式中: mb为冷冻干燥后核桃粕总蛋白的总质量, g; mc为冷冻干燥后核桃多肽的总质量, g
1.3.4 核桃多肽-硒螯合物的制备
称取一定质量的核桃多肽粉, 配制成浓度为4%的核桃多肽溶液, 调节pH至8, 水浴加热一定时间(使其达到反应温度), 水浴加热时需用封口膜使反应过程在封闭空间进行, 防止反应被干扰。再加入一定质量的亚硒酸钠, 恒温振荡反应, 用旋转蒸发仪进行蒸发浓缩, 待冷却至室温, 量取10倍体积无水乙醇加入反应体系, 静置12 h使螯合物沉降, 离心后收集沉淀, 真空冷冻干燥后即可制得核桃多肽-硒螯合物[8]。
(1)核桃多肽-硒螯合物制备的单因素研究
分别调整多肽与亚硒酸钠质量比、螯合温度、螯合时间和pH 4个因素水平, 探究表1中4个因素对硒螯合率的影响。各因素的基本前提条件为: 多肽与亚硒酸钠质量比1:1, 螯合温度50 ℃, pH 8, 螯合时间40 min。在考察某一因素对螯合率的影响时, 其他3个因素与基本条件保持一致。最后统一测定螯合物中的硒含量并计算硒螯合率。
(2)核桃多肽-硒螯合物制备的工艺优化研究
基于单因素试验结果, 采用4因素3水平正交试验法优化核桃多肽-硒螯合物制备工艺: 考察核桃多肽与亚硒酸钠质量比(A)、螯合pH (B)、螯合时间(C)和螯合温度(D)对螯合反应的影响。各因素水平设置如表2所示。
1.3.5 核桃多肽-硒螯合物硒含量及其螯合率的测定
(1)螯合物中的硒含量测定
参考王莲芳等[9]和WANG等[10]的方法进行硒含量测定。用适宜规格的移液管量取一定量硒标准溶液或适量样品, 稀释至40 mL, 将pH调至2, 加入2 mL EDTA-2Na溶液和2 mL 3,3'-二氨基联苯胺溶液, 混匀后避光反应50 min, 将pH调节至7, 处理好后置于分液漏斗中, 准确加入10 mL甲苯, 振摇2 min, 静置分层, 在分光光度计波长420 nm处测定甲苯层的吸光度[11]。
样品中硒含量的计算公式如公式(2)所示:
样品中硒的含量/(μg/g)=$\frac{CV}{WN}$
式中: C为从标准曲线上查得的相当于硒的标准质量浓度, µg/mL; V为甲苯萃取所得的样品体积, mL; W为样品的质量, g; N为用于测定的样品体积占定容后样品的体积分数, %。
(2)硒螯合率测定
采用1.3.5(1)方法测定螯合物中的硒含量, 硒螯合率计算公式如公式(3)所示:
硒螯合率/%=(m1/m2)×100%
式中: m1为螯合物中硒的总量, g; m2为反应体系中硒的总量, g。
1.3.6 核桃多肽-硒螯合物的紫外光谱分析
取相同质量的核桃多肽和核桃多肽-硒螯合物样品, 分别溶解于超纯水中, 制备成质量浓度为0.025 mg/mL的溶液。利用紫外分光光度计在波长范围为190~400 nm进行扫描, 以测定核桃多肽硒螯合物的紫外吸收光谱。
1.3.7 核桃多肽-硒螯合物的荧光光谱分析
分别称取一定质量的核桃多肽与核桃多肽-硒螯合物样品溶于超纯水中, 配制成0.025 mg/mL的溶液, 用荧光分光光度计进行扫描。设定仪器参数如下: 激发波长为280 nm, 发射波长范围为280~400 nm, 对核桃多肽-硒螯合物进行荧光光谱的测定。
1.3.8 核桃多肽-硒螯合物的溶解性测定
核桃多肽-硒螯合物溶解性的测定: 将样品分别配制成pH为2.0、4.0、6.0、8.0、10.0, 底物浓度为0.3%的溶液, 室温下搅拌30 min, 后将其装入规格为50 mL的离心管中进行离心(离心前配平), 设置离心机转速为5000 r/min, 离心2 min, 测定上清液中硒的含量。
1.3.9 核桃多肽-硒螯合物的热稳定性测定
将样品溶解于超纯水中, 配制成底物浓度为0.3%的标准溶液, 随后分别取20 mL溶液放入100 mL的锥形瓶中, 后将其分别置于40、60、80、100 ℃的恒温水浴锅中2 h, 然后冷却至到室温, 透析60 h后测定其硒含量。
1.3.10 核桃多肽-硒螯合物的酸碱稳定性测定
称取一定量的样品溶解于超纯水中, 经过精确稀释与混合, 制备成底物浓度为0.3%的标准溶液, 随后分别取20 mL溶液置于100 mL锥形瓶中, 使用1 mol/L的HCl和1 mol/L的NaOH调节各锥形瓶中的溶液的pH至3、5、7、9、11, 随后将调节完pH后的溶液放于37 ℃恒温水浴锅中放置2 h, 以确保反应充分进行, 透析60 h后测定其硒含量。
1.3.11 核桃多肽-硒螯合物的体外模拟胃肠道消化试验
(1)胃蛋白酶消化试验
称取一定量的样品溶解于超纯水中, 经过精确稀释与混合, 制备成底物浓度为0.3%的标准溶液, 取其中30 mL溶液于100 mL锥形瓶中。使用1 mol/L的HCI调节溶液pH至2.0, 在37 ℃恒温水浴中孵育20 min。在此后添加0.6%(物料比, 质量分数)的胃蛋白酶, 充分搅拌均匀, 后将溶液置于37 ℃恒温水浴中4 h, 最后置于沸水浴中沸煮20 min以终止酶解反应, 透析60 h后测定其硒含量[12]。
(2)胰蛋白酶消化试验
称取一定量的样品溶解于超纯水中, 经过精确稀释与混合, 制备成底物浓度为0.3%的标准溶液,取其中30 mL溶液于锥形瓶中。使用10%的NaOH调节溶液的pH至7.5, 后置于37 ℃恒温水浴中孵育20 min。添加2%(物料比, 质量分数)的胰蛋白酶, 充分搅拌均匀, 后将溶液置于37 ℃恒温水浴中处理6 h, 最后于沸水浴中沸煮20 min以终止酶解反应, 透析60 h后测定其硒含量。
1.3.12 核桃多肽-硒螯合物的抗氧化活性测定
(1) DPPH自由基清除率的测定
称取适量核桃多肽-硒螯合物, 分别配制成质量浓度为0.1、0.2、0.3、0.4、0.5、0.6 mg/mL的样品溶液, 参考蔡俊等[13]的研究方法测定DPPH自由基清除率。用同样的方法重复测定3次, 计算其平均值, 并将核桃多肽与螯合物清除能力进行对比。可按公式(4)计算DPPH自由基清除率:
DPPH自由基清除率/%=$\left(1-\frac{{A}_{\text{i}}-{A}_{j}}{{A}_{0}}\right)\times 100\%$
式中: Ai为样品吸光度; Aj为等体积无水乙醇吸光度; A0为等体积超纯水吸光度。
(2)羟基自由基清除率的测定
参考林栋等[14]的研究方法进行操作, 称取适量核桃多肽-硒螯合物, 分别配制成质量浓度为0.1、0.2、0.3、0.4、0.5、0.6 mg/mL的样品溶液, 取不同质量浓度的样品溶液1.0 mL置于小烧杯中, 按顺序加入硫酸亚铁溶液、水杨酸乙醇溶液和过氧化氢溶液各1 mL, 37 ℃反应0.5 h, 在紫外可见分光光度计波长510 nm处测定样品吸光度用同样的方法重复测定3次, 计算其平均值, 并将核桃多肽与螯合物清除能力进行对比。可按公式(5)计算羟基自由基清除率:
羟基自由基清除率/%=$\left(1-\frac{{B}_{\text{i}}-{B}_{j}}{{B}_{0}}\right)\times 100\%$
式中: Bi为样品吸光度; Bj为等体积无水乙醇吸光度; B0为等体积超纯水吸光度。
(3) ABTS阳离子自由基清除率的测定
参考张榕欣等[15]的测定方法配制ABTS工作液。称取适量核桃多肽-硒螯合物, 分别配制成质量浓度为0.1、0.2、0.3、0.4、0.5、0.6 mg/mL的样品溶液, 参考蔡俊等[13]的研究方法测定ABTS阳离子自由基清除率。用同样的方法重复测定3次, 计算其平均值, 并将核桃多肽与螯合物清除能力进行对比。可按公式(6)计算ABTS阳离子自由基清除率:
ABTS阳离子自由基清除率/%=$\left(1-\frac{{C}_{\text{i}}-{C}_{j}}{{C}_{0}}\right)\times 100\%$
式中: Ci为样品吸光度; Cj为等体积无水乙醇吸光度; C0为等体积超纯水吸光度
1.4 数据处理
每组进行3次平行试验, 结果以平均值±标准偏差表示。使用GraphPad Prism 6.0软件对数据进行统计分析和绘图, 用单因素方差分析分析组间显著性差异。用小写字母表示5%水平上的显著性差异。
2 结果与分析
收起
2.1 核桃多肽硒螯合物的制备结果
2.1.1 单因素试验结果分析
由图1A可见, 随着多肽与亚硒酸钠质量比的增加, 核桃多肽-硒螯合物的螯合率呈现出一个先升后降的趋势。在多肽与亚硒酸钠的质量比为3:1时螯合率达到峰值51.04%。这一现象可能是因为在一定范围内, 随着多肽量的增加, 有更多的多肽分子与亚硒酸钠中的硒离子结合, 从而使螯合率上升。当多肽与亚硒酸钠质量比超过3:1后, 可能由于体系中过多的多肽会相互聚集, 减少了与硒离子接触的有效活性位点, 导致螯合率下降[16]。因此, 确定多肽与亚硒酸钠的质量比为3:1。
图1B显示螯合物的螯合率受温度变化影响显著, 呈现先升后缓降的趋势, 最佳螯合温度为50 ℃。在温度低于50 ℃时, 随着温度升高, 分子热运动加剧, 多肽分子和硒离子的碰撞频率增加, 使得两者更容易结合形成螯合物, 所以螯合率与温度呈正相关。而当温度超过50 ℃时, 硒螯合率缓慢下降。这是因为温度过高会破坏核桃多肽的空间结构, 使其变性失活。多肽的活性中心结构被破坏后, 就无法有效地与硒离子发生螯合反应, 导致螯合率降低[17-19]。因此, 确定最佳螯合温度为50 ℃, 以保证螯合反应的顺利进行, 同时避免因温度过高导致多肽变性带来的损失。
由图1C可见, 螯合时间的变化对硒螯合率的影响并不剧烈, 但也呈现出一定规律。当螯合时间小于等于50 min, 螯合率平稳增长, 这可能是因为随着时间的延长, 多肽和硒离子有更多的时间相互接触和反应, 从而使螯合反应不断进行, 螯合率逐渐提高。当大于50 min, 螯合率却缓缓下降。这可能是由于核桃多肽-硒螯合物本身不稳定, 过长的螯合时间会导致已生成的螯合物分解, 从而降低螯合率有关[20]。因此, 选择50 min作为最佳螯合时间, 可以在保证较高螯合率的同时, 提高生产效率, 避免不必要的时间浪费。
图1D表明pH的变化对硒螯合率的影响颇为显著, 最佳螯合pH为8。当pH低于8时, 硒螯合率随着pH的增大而逐渐上升; 当pH低于8时, 硒螯合率随着pH的增大而逐渐上升, 在pH为8的条件下, 硒盐中的硒离子更易于电离, 从而有利于螯合物的生成[14]。这是因为合适的pH环境能够改变多肽分子的带电状态, 使其活性基团更有利于与硒离子结合。当pH超过8时, 硒螯合率开始逐渐下降, 可能是因为过高的pH会导致多肽分子结构发生变化, 影响其与硒离子的结合能力, 同时也可能会使硒离子发生其他化学反应, 降低了参与螯合反应的硒离子浓度[21]。因此, 可以确定最佳的螯合pH为8。
2.1.2 正交试验结果分析
以硒螯合率为指标, 得出的正交试验结果见表3。
根据表3的结果, 各因素对硒螯合率的影响程度排序为: 核桃多肽与亚硒酸钠质量比(A)>pH (B)>螯合时间(C)>螯合温度(D)。通过正交试验, 确定了最优组合为A2B2C2D1, 即多肽与亚硒酸钠质量比为3:1, 螯合pH为8, 螯合温度为45 ℃, 螯合时间为50 min, 此时硒螯合率最高, 达到了58.99%±1.44%, 较林栋等[14]报道的方案(多肽浓度4%、pH 8、50 ℃、质量比3:1、50 min)提升8.18%。
2.2 核桃多肽-硒螯合物的紫外光谱结果
根据紫外光谱显示, 金属离子与配位体结合确实能改变配体的电子跃迁特性。通过对比核桃多肽-硒螯合物与核桃多肽的吸收峰变化, 可以清晰地看到两者在最大吸收波长位置和吸收强度上的差异。如图2所示, 核桃多肽-硒螯合物在波长约200~210 nm处有一个明显的强吸收峰。在紫外光谱中, 200~220 nm附近的吸收峰通常与肽键的π-π*跃迁有关, 这表明核桃多肽-硒螯合物中存在肽键结构, 且该结构在紫外光区有特征吸收。与核桃多肽相比, 螯合物在该强吸收峰处的吸收强度更高, 可能是由于硒的螯合作用影响了肽键周围的电子云分布, 从而增强了该波长下的吸收[22]。核桃多肽-硒螯合物在200~210 nm处的吸收强度明显高于其他波长范围, 随着波长的增加, 吸收强度逐渐下降, 在250~300 nm之间有一些小的波动, 但幅度较小。螯合物的光谱形状与核桃多肽的光谱有一定差异, 说明硒的螯合改变了核桃多肽原本的电子结构和分子构型, 进而影响了其对不同波长紫外光的吸收特性。通过与核桃多肽紫外光谱对比, 进一步证明了硒螯合作用对核桃多肽结构和光学性质产生了影响[23]。
2.3 核桃多肽-硒螯合物的荧光光谱结果
芳香族氨基酸所产生的荧光变化, 作为一种间接指标, 能够有效地揭示多肽结构的变化情况。在300 nm和350 nm波长处观察到的吸收峰, 分别对应着色氨酸和酪氨酸的特征吸收[24]。图3所示的荧光光谱图清晰地表明, 核桃多肽-硒螯合物在最强吸收峰位置和强度上, 虽然变化幅度不大, 但始终低于核桃多肽。综合来看, 硒与核桃多肽螯合后螯合物的荧光强度降低, 光谱形状也发生了细微而明确的变化。这些变化暗示了硒螯合作用对核桃多肽分子结构和电子状态的修饰, 有助于进一步探究其在生物体内的作用机制和功能特性。
2.4 核桃多肽-硒螯合物的溶解性结果
图4揭示了不同pH对核桃多肽-硒螯合物中硒溶解性的影响。从图4中可以清晰看出, 当pH小于或等于8时, 核桃多肽-硒螯合物在水中的溶解度逐渐上升; 而当pH超过8时, 其溶解度则明显下降。这一变化表明, 碱性条件可能促使多肽链发生变构或聚合, 从而导致溶解性降低[25]。另外, 在极端酸性环境(pH=2)下, 核桃多肽-硒螯合物的溶解度达到了最低点, 可能是由于强酸环境导致Se-O/N键质子化断裂[26]。
2.5 核桃多肽-硒螯合物的热稳定性结果
图5揭示了在不同热处理条件下对核桃多肽-硒螯合物中的硒含量的影响。从图5中数据可以看出, 在经过2 h的连续加热处理后, 核桃多肽-硒螯合物的硒含量并未出现显著变化, 这充分说明了其整体稳定性极佳。特别是在60~100 ℃的温度范围内, 硒含量虽有轻微波动, 但变化并不明显。值得注意的是, 即便在20 ℃的较低温度下, 硒含量也仅是略微降低, 但仍保持在94%以上的高水平。这一结果有力地证实了硒离子与多肽之间形成的化学键合具有出色的耐热性, 能够在高温条件下保持结构的稳定, 从而确保了核桃多肽-硒螯合物在较高温度下的结构与功能完整性。
2.6 核桃多肽-硒螯合物的酸碱稳定性结果
图6展示的试验结果揭示了在不同pH条件下, 经37 ℃处理2 h的核桃多肽-硒螯合物中硒含量的变化趋势。随着溶液pH的逐步增大, 硒含量展现出一个先上升后下降的过程。特别地, 在pH为7时, 硒含量达到了一个峰值, 高达86.74%。即便在酸性较强的pH=3的环境下, 硒含量虽有所下降, 但仍维持在77%以上的较高水平。这一结果证明了, 核桃多肽-硒螯合物在广泛的pH条件下, 无论是酸性还是碱性介质, 均能展现出较强的稳定性, 其硒含量保持在一个相对较高的水平。
2.7 核桃多肽-硒螯合物的体外模拟胃肠道消化试验结果
图7揭示了不同消化方式的作用下对核桃多肽-硒螯合物中硒含量的影响, 从图7中数据可以看出, 经过胃蛋白酶与胰蛋白酶的体外消化处理后, 核桃多肽-硒螯合物中的硒含量相对比例均维持在90%以上, 这有力地证明了核桃多肽与硒之间配位结合的高度稳定性[27]。即便在肽链遭受酶促断裂的情况下, 这种结合依然保持稳固[28]。这一结果充分表明, 核桃多肽-硒螯合物在模拟的消化过程中展现出了良好的稳定性, 能够有效抵御消化酶的降解作用[29]。
2.8 核桃多肽-硒螯合物的抗氧化活性结果
ZHANG等[30]研究表明, 核桃多肽具有良好的抗氧化活性, 能够提高一些抗氧化酶的水平, 从而有效降低自由基对HT22细胞的氧化损伤。方热军等[31]研究表明, 硒是多种酶的活性中心, 可以参与生物机体内多种抗氧化酶的合成, 其抗氧化能力主要体现于含硒酶。
由图8可知, 核桃多肽-硒螯合物在对DPPH自由基的清除能力随质量浓度的增加显著提升, 核桃多肽-硒螯合物的清除能力均强于核桃多肽, 表明螯合后抗氧化活性得到增强。图9显示, 两者清除羟基自由基的能力均随质量浓度增加而增强, 但核桃多肽-硒螯合物的增强趋势更为明显, 尤其在质量浓度超过0.30 mg/mL后, 其清除率数据(0.50 mg/mL浓度下50.52%±0.63%)与林栋等[14]报道的同类研究结果趋势一致。图10则表明, 在0.10 mg/mL质量浓度时, 两者清除ABTS阳离子自由基的能力相当, 但在0.20~0.60 mg/mL质量浓度范围内, 核桃多肽-硒螯合物的清除能力显著强于核桃多肽。综上所述, 核桃多肽与亚硒酸钠螯合后, 其抗氧化活性得到显著提升。综上所述, 核桃多肽-硒螯合物展现出显著的自由基清除能力。
从试验结果可以推测, 硒与核桃多肽的螯合作用改变了核桃多肽的化学结构和物理性质, 进而影响了其抗氧化活性。可能的机制包括: 硒原子的引入改变了核桃多肽分子的电子云分布, 使其更容易提供电子给自由基, 从而终止自由基链式反应[32]; 硒可能与核桃多肽中的某些氨基酸残基形成了新的化学键或相互作用, 改变了多肽分子的空间构象, 使得活性基团更易于暴露和与自由基反应[32]; 此外, 硒本身也可能具有一定的抗氧化能力, 与核桃多肽协同作用, 增强了整体的抗氧化效果[33]。由于核桃多肽-硒螯合物具有显著增强的抗氧化活性, 其在食品、保健品和医药等领域具有潜在的应用价值。
3 结论
收起
本研究以核桃多肽和亚硒酸钠为原材料, 制备核桃多肽-硒螯合物, 并对其溶解性、热稳定性、酸碱稳定性及体外消化特性和抗氧化活性进行了深入探究。核桃多肽-硒螯合物的最佳制备条件为肽硒质量比3:1、螯合温度45 ℃、时间50 min、pH为8, 此时硒螯合率达到58.99%±1.44%。紫外扫描光谱显示, 螯合反应后样品的最大吸收波长和吸收强度均发生变化, 表明其为一种新物质。荧光光谱分析进一步揭示, 核桃多肽与硒离子的配位结合导致内部基团变化, 引发荧光猝灭。核桃多肽-硒螯合物具有良好的水溶性和热稳定性, 经酶解后硒含量变化较小。核桃多肽与核桃多肽-硒螯合物均具备抗氧化活性, 且螯合物的抗氧化活性明显提升, 能减轻自由基造成的氧化损伤。因此, 核桃多肽-硒螯合物有望成为用于低硒人群的功能性营养食品原料, 不仅拓展了核桃多肽在食品领域的应用, 还为开发补硒功能食品原料提供了理论依据, 并为硒缺乏相关慢性疾病的治疗提供了新方向。
基金
收起
  • 陕西学前师范学院引进人才(博士)科研启动项目(2023DS02)
  • 陕西高校青年创新团队“秦岭药食同源生物资源保护与利用创新团队”项目
  • 陕西学前师范学院科研创新团队“陕西特色农产品精深加工技术创新团队”项目
  • “儿童健康照明LED荧光粉”项目
  • 西安市科技计划项目(23NYGG0061)
  • 陕西省体育局常规课题项目(20240468)
文献
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2025年第16卷第15期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250121005
  • 接收时间：2025-01-21
  • 首发时间：2026-01-09
  • 出版时间：2025-08-15
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  • 收稿日期：2025-01-21
基金
陕西学前师范学院引进人才(博士)科研启动项目(2023DS02)
陕西高校青年创新团队“秦岭药食同源生物资源保护与利用创新团队”项目
陕西学前师范学院科研创新团队“陕西特色农产品精深加工技术创新团队”项目
“儿童健康照明LED荧光粉”项目
西安市科技计划项目(23NYGG0061)
陕西省体育局常规课题项目(20240468)
作者信息
    陕西学前师范学院生物食品与化学学院, 西安 710100

通讯作者:

*王静(1985—), 女, 博士, 副教授, 主要研究方向为食品功能因子的研究与开发。E-mail:
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2种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
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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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