Article(id=1320889619059687599, tenantId=1146029695717560320, journalId=1260987677001138203, issueId=1320889172802515656, articleNumber=null, orderNo=null, doi=10.13386/j.issn1002-0306.2025030090, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1741536000000, receivedDateStr=2025-03-10, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1792854226125, onlineDateStr=2026-10-24, pubDate=1768406400000, pubDateStr=2026-01-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792854226125, onlineIssueDateStr=2026-10-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792854226125, creator=13701087609, updateTime=1792854226125, updator=13701087609, issue=Issue{id=1320889172802515656, tenantId=1146029695717560320, journalId=1260987677001138203, year='2026', volume='47', issue='2', pageStart='1', pageEnd='514', issueExtLink='null', onlineDate='null', pubDate='1768406400000', pubDateStr='2026-01-15', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792854119730, creator='13701087609', updateTime=1792854119730, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=277, endPage=286, ext={EN=ArticleExt(id=1320889619261014192, articleId=1320889619059687599, tenantId=1146029695717560320, journalId=1260987677001138203, language=EN, title=Optimization of α-Linolenic Acid Enrichment Process and Quality Analysis in Germinated Flaxseeds, columnId=1261343843090313538, journalTitle=Science and Technology of Food Industry, columnName=Processing Technology, runingTitle=null, highlight=null, articleAbstract=

In order to develop functional sprouted foods rich in α-linolenic acid (ALA), this study investigated the effects of incubation temperature, salt concentration, and pH on the ALA content during the sprouting process using flaxseed as the raw material. Based on the results of single-factorial experiments, Box-Behnken response surface methodology was used to optimize the sprouting conditions, and the quality and bioactivity of sprouted flaxseed were comprehensively evaluated. The results indicated that the optimal germination time was 48 h, and the optimized process parameters were an incubation temperature of 26 ℃, a salt concentration of 0.5 mg/mL NaCl, and a pH of 6.0, resulting in an ALA content of 20.42 g/100 g. Under these conditions, the activities of lipoxygenase (LOX), allene oxide synthase (AOS), and allene oxide cyclase (AOC) were significantly enhanced (P<0.05). Meanwhile, the expression of fad2, fad3, fad7, and fad8 genes was significantly upregulated (P<0.05), while the expression of lox gene was downregulated (P<0.05), thereby promoting the synthesis and accumulation of ALA. When flax seeds were sprouted for different periods of time under the optimal sprouting conditions, it was found that the sprouted flax seeds had better nutritional value compared with the unsprouted ones, with the ALA content reaching 24.125 g/100 g (P<0.05), an increase of 13.125 g/100 g. The protein content was increased to 25.75% (P<0.05), the lignan content was increased by 1.29 mg/mL (P<0.05), and the γ-aminobutyric acid (GABA) content was increased by 28.99% (P<0.05), while the cyanogenic glycoside content was reduced by 97.62% (P<0.05). In addition, bioactivity was significantly enhanced (P<0.05), with superoxide dismutase (SOD) activity increasing by 154.25 U/mL (P<0.05). The antioxidant capacity was also significantly improved, with the DPPH and ABTS+ free radical scavenging rates increasing by 12.51% and 41.67%, respectively (P<0.05), and the ferrous ion reducing power reaching 0.88 mmol/L (P<0.05). In conclusion, the biosynthesis and accumulation of ALA in flaxseed were significantly improved by optimized germination conditions, and its nutritional quality and bioactivity were enhanced, providing theoretical basis and technical support for the development of healthy foods rich in ALA.

, authors=Rongjun HE1, *, Yu YANG1, Jinhao ZHANG1, Xiushuang LI2, Xinru LIU1, Wei WANG2, authorsList=Rongjun HE, Yu YANG, Jinhao ZHANG, Xiushuang LI, Xinru LIU, Wei WANG, authorCompany=null, correspAuthors=Rongjun HE, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright © 2026 Science and Technology of Food Industry. All rights reserved., 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, fund=null), CN=ArticleExt(id=1320889622805201093, articleId=1320889619059687599, tenantId=1146029695717560320, journalId=1260987677001138203, language=CN, title=萌芽亚麻籽中α-亚麻酸的富集工艺优化及品质分析, columnId=1261343845481066826, journalTitle=食品工业科技, columnName=工艺技术, runingTitle=null, highlight=null, articleAbstract=

为开发富含α-亚麻酸(α-Linolenic Acid,ALA)的功能性萌芽食品,本研究以亚麻籽为原料,探究培养温度、盐浓度和pH对萌芽过程中ALA含量的影响。基于单因素实验结果,采用Box-Behnken响应面法优化培养条件,并对萌芽亚麻籽的品质及生物活性进行综合评价。结果表明,最佳萌芽时间为48 h,优化工艺参数为培养温度26 ℃、盐浓度0.5 mg/mL NaCl和pH6.0,ALA含量为20.42 g/100 g。在此条件下,脂氧合酶(Lipoxygenase,LOX)、丙二烯氧化物合成酶(Allene oxide synthase,AOS)和丙二烯氧化物环化酶(Allene oxide cyclase,AOC)的活性显著提高(P<0.05)。同时,fad2、fad3、fad7和fad8基因的表达显著上调(P<0.05),而lox基因的表达显著下调(P<0.05),从而促进了ALA的合成与积累;在最佳萌芽工艺条件下培养不同萌芽时间亚麻籽,发现与未萌芽的相比,萌芽的亚麻籽具有良好的营养价值,ALA含量达24.125 g/100 g(P<0.05),增加了13.125 g/100 g,蛋白质含量提升至25.75%(P<0.05),木酚素含量提升1.29 mg/mL(P<0.05),γ-氨基丁酸(γ-Aminobutyric acid,GABA)含量提升了28.99%(P<0.05),生氰糖苷含量降低97.62%(P<0.05);同时,生物活性显著增强(P<0.05),超氧化物歧化酶(Superoxide dismutase,SOD)活性提高154.25 U/mL(P<0.05);抗氧化能力显著改善,DPPH和ABTS+自由基清除率分别提高了12.51%和41.67%(P<0.05),铁离子还原能力达0.88 mmol/L(P<0.05)。综上所述,通过优化萌芽条件显著提升了亚麻籽中ALA的生物合成与积累,同时改善了其营养品质和生物活性,为富含ALA的健康食品研发提供了理论依据和技术支持。

, authors=何荣军1, *, 杨雨1, 张锦豪1, 李秀双2, 刘芯如1, 王威2, authorsList=何荣军, 杨雨, 张锦豪, 李秀双, 刘芯如, 王威, authorCompany=null, correspAuthors=何荣军, authorNote=

何荣军(1978−),男,硕士,副教授,研究方向:功能性食品开发,E-mail:

, correspAuthorsNote=
何荣军(1978−),男,硕士,副教授,研究方向:功能性食品开发,E-mail:。
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何荣军(1978−),男,硕士,副教授,研究方向:功能性食品开发,E-mail:

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何荣军(1978−),男,硕士,副教授,研究方向:功能性食品开发,E-mail:

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注:图中不同小写字母表示差异显著(P<0.05);图3~图5、图7~图9同。

, figureFileSmall=wlaAVIdA6/0cZnUDQm8FyQ==, figureFileBig=tYitblyFMB4mVZA4HHkC7w==, tableContent=null), ArticleFig(id=1320889627628650742, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Fig.2, caption=Response surface plots of the interaction effects of two factors on ALA content in sprouted flaxseeds, figureFileSmall=yu2U5lmHcsklo+lMxo4PZw==, figureFileBig=fnnN+u39jQZUlNCrQOi3zQ==, tableContent=null), ArticleFig(id=1320889627699953911, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=图2, caption=两因素交互作用对萌芽亚麻籽ALA含量影响的曲面图, figureFileSmall=yu2U5lmHcsklo+lMxo4PZw==, figureFileBig=fnnN+u39jQZUlNCrQOi3zQ==, tableContent=null), ArticleFig(id=1320889627767062776, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Fig.3, caption=Effects of temperature, salt concentration, and pH variation on enzyme activity, figureFileSmall=x0kxwPcWueYiXPNaOur7xg==, figureFileBig=aPZAgex+ppk/WsKPs2BxHg==, tableContent=null), ArticleFig(id=1320889627825783033, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=图3, caption=温度、盐浓度、pH变化对酶活性的影响, figureFileSmall=x0kxwPcWueYiXPNaOur7xg==, figureFileBig=aPZAgex+ppk/WsKPs2BxHg==, tableContent=null), ArticleFig(id=1320889627888697594, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Fig.4, caption=Gene expression of key enzymes at different ALA levels, figureFileSmall=7pg0qcLPkpfyII3LY83g3w==, figureFileBig=cPkii0zEZrTGTLIcTHCK0g==, tableContent=null), ArticleFig(id=1320889627947417851, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=图4, caption=不同ALA含量中关键酶基因的相对表达量, figureFileSmall=7pg0qcLPkpfyII3LY83g3w==, figureFileBig=cPkii0zEZrTGTLIcTHCK0g==, tableContent=null), ArticleFig(id=1320889628006138108, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Fig.5, caption=Changes in fundamental physicochemical indices during flaxseed germination, figureFileSmall=RA1IXz4fkDy6j+EATVKkaQ==, figureFileBig=Qp3bD1OgV/Ois26St290pw==, tableContent=null), ArticleFig(id=1320889628069052669, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=图5, caption=亚麻籽萌芽过程中基本理化指标变化, figureFileSmall=RA1IXz4fkDy6j+EATVKkaQ==, figureFileBig=Qp3bD1OgV/Ois26St290pw==, tableContent=null), ArticleFig(id=1320889628144550142, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Fig.6, caption=Changes in fatty acid composition during flaxseed germination, figureFileSmall=g1oWJ/PM0vvpJvZmVYjWPA==, figureFileBig=Gj9Sk15t2Le5lFg8HLeb9A==, tableContent=null), ArticleFig(id=1320889628211659007, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=图6, caption=亚麻籽萌芽过程中脂肪酸组成成分的变化, figureFileSmall=g1oWJ/PM0vvpJvZmVYjWPA==, figureFileBig=Gj9Sk15t2Le5lFg8HLeb9A==, tableContent=null), ArticleFig(id=1320889628282962176, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Fig.7, caption=Changes in lignan content during flaxseed germination, figureFileSmall=PziX3dCKL8LWONqx2FpgEg==, figureFileBig=Mm4zjmQmoRJoXcYlp62cHA==, tableContent=null), ArticleFig(id=1320889628371042561, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=图7, caption=亚麻籽萌芽过程中木酚素含量的变化, figureFileSmall=PziX3dCKL8LWONqx2FpgEg==, figureFileBig=Mm4zjmQmoRJoXcYlp62cHA==, tableContent=null), ArticleFig(id=1320889628442345730, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Fig.8, caption=Changes in GABA content during flaxseed germination, figureFileSmall=BJY8zPHxOV9bcd5Toq7Crw==, figureFileBig=CV8RxK8JLEuUZ3N8SoEIdQ==, tableContent=null), ArticleFig(id=1320889628509454595, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=图8, caption=亚麻籽萌芽过程中GABA含量的变化, figureFileSmall=BJY8zPHxOV9bcd5Toq7Crw==, figureFileBig=CV8RxK8JLEuUZ3N8SoEIdQ==, tableContent=null), ArticleFig(id=1320889628572369156, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Fig.9, caption=Changes in cyanogenic glycoside content during flaxseed germination, figureFileSmall=cldgD6tgrGDUMbSDQkd1ZA==, figureFileBig=Xod8+dg6ObMRHZeSZoDxPg==, tableContent=null), ArticleFig(id=1320889628647866629, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=图9, caption=亚麻籽萌芽过程中生氰糖苷含量的变化, figureFileSmall=cldgD6tgrGDUMbSDQkd1ZA==, figureFileBig=Xod8+dg6ObMRHZeSZoDxPg==, tableContent=null), ArticleFig(id=1320889628727558406, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Fig.10, caption=Changes in antioxidant capacity during flaxseed sprouting, figureFileSmall=JXZ9KMIIU3g6aa9nyQrFDA==, figureFileBig=qnxc6RtY77v66BZXEvRN9g==, tableContent=null), ArticleFig(id=1320889628811444487, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=图10, caption=亚麻籽萌芽过程中抗氧化能力的变化

注:图中不同小写字母表示相同萌芽时间不同浓度之间的差异显著(P<0.05)。

, figureFileSmall=JXZ9KMIIU3g6aa9nyQrFDA==, figureFileBig=qnxc6RtY77v66BZXEvRN9g==, tableContent=null), ArticleFig(id=1320889628882747656, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Table 1, caption=

Factors and levels of the Box-Behnken experimental design

, figureFileSmall=null, figureFileBig=null, tableContent=
因素编码水平
−101
培养温度(℃)A202530
盐浓度(mg/mL)B0.30.50.7
pHC5.06.07.0
), ArticleFig(id=1320889628962439433, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=表1, caption=

Box-Behnken响应面试验因素水平设计

, figureFileSmall=null, figureFileBig=null, tableContent=
因素编码水平
−101
培养温度(℃)A202530
盐浓度(mg/mL)B0.30.50.7
pHC5.06.07.0
), ArticleFig(id=1320889629016965386, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Table 2, caption=

PCR primer sequence

, figureFileSmall=null, figureFileBig=null, tableContent=
基因正向引物(5’-3’)反向引物(5’-3’)
fad2TGGATTCAGGTGGT
GCTGTC
GGCTCTCATCCGCG
ATAACA
fad3GCGTCATTCCAGCCC
TATCA
GACTCCGCACCAAG
AAGTGA
fad7AAGTGCACCGCTTA
GGATCC
TATGCCAGCAAGGG
AAAGGG
fad8GCCAATGTCATGGTGGA
TATTATT
GCGGCAAGGAATGGAA
TTATC
loxACGTCATCAAAACATG
CACCA
CAGAGGTTGGGTATCC
GCAGT
β-ACTINGCCATCTTTGATTG
GAATGG
GGTGCCACAACCTTG
ATCTT
), ArticleFig(id=1320889629121822987, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=表2, caption=

PCR引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=
基因正向引物(5’-3’)反向引物(5’-3’)
fad2TGGATTCAGGTGGT
GCTGTC
GGCTCTCATCCGCG
ATAACA
fad3GCGTCATTCCAGCCC
TATCA
GACTCCGCACCAAG
AAGTGA
fad7AAGTGCACCGCTTA
GGATCC
TATGCCAGCAAGGG
AAAGGG
fad8GCCAATGTCATGGTGGA
TATTATT
GCGGCAAGGAATGGAA
TTATC
loxACGTCATCAAAACATG
CACCA
CAGAGGTTGGGTATCC
GCAGT
β-ACTINGCCATCTTTGATTG
GAATGG
GGTGCCACAACCTTG
ATCTT
), ArticleFig(id=1320889629205709068, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Table 3, caption=

Box-Behnken experimental design and results

, figureFileSmall=null, figureFileBig=null, tableContent=
实验号ABCY(g/100 g)
1−1−1013.52
21−1015.46
3−11014.44
411015.89
5−10−111.71
610−112.86
7−10113.09
810115.06
90−1−112.11
1001−112.50
110−1113.92
1201114.57
1300016.88
1400017.34
1500017.90
1600017.80
1700018.03
), ArticleFig(id=1320889629285400845, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=表3, caption=

Box-Behnken试验设计与结果

, figureFileSmall=null, figureFileBig=null, tableContent=
实验号ABCY(g/100 g)
1−1−1013.52
21−1015.46
3−11014.44
411015.89
5−10−111.71
610−112.86
7−10113.09
810115.06
90−1−112.11
1001−112.50
110−1113.92
1201114.57
1300016.88
1400017.34
1500017.90
1600017.80
1700018.03
), ArticleFig(id=1320889629356704014, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Table 4, caption=

Regression ANOVA

, figureFileSmall=null, figureFileBig=null, tableContent=
项目平方和自由度均方F值Prob>F显著性
模型71.8197.9859.88<0.0001**
A-培养温度5.3015.3039.760.0004**
B-盐浓度0.714010.71405.360.0432*
C-pH6.9616.9652.200.0002**
AB0.060010.06000.45050.5236
AC0.168110.16811.260.2984
BC0.016910.01690.12680.7322
A28.6018.6064.50<0.0001**
B27.4917.4956.210.0001**
C237.42137.42280.84<0.0001**
残差0.932870.1333
失拟项0.032430.01080.04790.9842
净误差0.900440.2251
总离差72.7416
), ArticleFig(id=1320889629432201487, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=表4, caption=

回归方差分析

, figureFileSmall=null, figureFileBig=null, tableContent=
项目平方和自由度均方F值Prob>F显著性
模型71.8197.9859.88<0.0001**
A-培养温度5.3015.3039.760.0004**
B-盐浓度0.714010.71405.360.0432*
C-pH6.9616.9652.200.0002**
AB0.060010.06000.45050.5236
AC0.168110.16811.260.2984
BC0.016910.01690.12680.7322
A28.6018.6064.50<0.0001**
B27.4917.4956.210.0001**
C237.42137.42280.84<0.0001**
残差0.932870.1333
失拟项0.032430.01080.04790.9842
净误差0.900440.2251
总离差72.7416
), ArticleFig(id=1320889629499310352, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=EN, label=Table 5, caption=

Changes in amino acid composition during flaxseed germination

, figureFileSmall=null, figureFileBig=null, tableContent=
氨基酸种类氨基酸含量(mg/g)
0 h24 h48 h72 h96 h
天冬氨酸4.1±0.21d5.4±0.35c9.4±0.07b12.4±0.35a11.4±0.84a
丝氨酸5.9±0.35e14.6±0.16d48.6±0.72c58.6±0.21b30.5±0.84a
谷氨酸6.0±0.42d13.1±0.53c22.9±0.19a21.8±0.10a18.3±0.43b
甘氨酸1.1±0.21c2.11±0.37c6.6±0.25b10.4±0.77a5.0±0.43b
丙氨酸3.2±0.31e6.6±0.12d14.7±0.74b16.8±0.43a8.6±0.21c
胱氨酸0.8±0.11b0.7±0.16b2.3±0.24a2.3±0.15a2.2±0.53a
缬氨酸3.7±0.04d5.2±0.51c13.7±0.34ab14.0±0.43a12.4±0.38b
蛋氨酸2.0±0.64b2.7±0.57b6.7±0.15a6.1±0.36a5.5±0.28a
异亮氨酸2.8±0.56d4.1±0.44c10.9±0.36a11.5±0.52a8.4±0.36b
亮氨酸4.8±0.41d6.3±0.65c18.6±0.89a17.4±0.41a15.3±0.29b
酪氨酸2.6±0.04e3.8±0.11d11.1±0.85a10.6±0.17b8.6±0.19c
苯丙氨酸3.7±0.53d5.4±0.10c16.6±0.12a16.6±0.47a14.2±0.55b
组氨酸6.0±0.43c3.2±0.31d18.5±0.76b21.9±0.68a20.1±0.22a
赖氨酸2.3±0.09e3.2±0.31d9.9±0.28b10.8±0.08a9.5±0.20c
精氨酸3.8±0.24d6.9±0.33c24.9±0.52b29.8±0.50a29.2±0.51a
苏氨酸1.4±0.04d2.2±0.12c4.6±0.54a4.3±0.05a3.4±0.64b
色氨酸0.6±0.65c0.8±0.06c1.9±0.02a1.8±0.05a1.5±0.10b
脯氨酸1.7±0.16e4.4±0.38d22.3±0.80c51.3±0.43a35.5±0.10b
), ArticleFig(id=1320889629574807825, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889619059687599, language=CN, label=表5, caption=

亚麻籽萌芽过程中氨基酸组成的变化

, figureFileSmall=null, figureFileBig=null, tableContent=
氨基酸种类氨基酸含量(mg/g)
0 h24 h48 h72 h96 h
天冬氨酸4.1±0.21d5.4±0.35c9.4±0.07b12.4±0.35a11.4±0.84a
丝氨酸5.9±0.35e14.6±0.16d48.6±0.72c58.6±0.21b30.5±0.84a
谷氨酸6.0±0.42d13.1±0.53c22.9±0.19a21.8±0.10a18.3±0.43b
甘氨酸1.1±0.21c2.11±0.37c6.6±0.25b10.4±0.77a5.0±0.43b
丙氨酸3.2±0.31e6.6±0.12d14.7±0.74b16.8±0.43a8.6±0.21c
胱氨酸0.8±0.11b0.7±0.16b2.3±0.24a2.3±0.15a2.2±0.53a
缬氨酸3.7±0.04d5.2±0.51c13.7±0.34ab14.0±0.43a12.4±0.38b
蛋氨酸2.0±0.64b2.7±0.57b6.7±0.15a6.1±0.36a5.5±0.28a
异亮氨酸2.8±0.56d4.1±0.44c10.9±0.36a11.5±0.52a8.4±0.36b
亮氨酸4.8±0.41d6.3±0.65c18.6±0.89a17.4±0.41a15.3±0.29b
酪氨酸2.6±0.04e3.8±0.11d11.1±0.85a10.6±0.17b8.6±0.19c
苯丙氨酸3.7±0.53d5.4±0.10c16.6±0.12a16.6±0.47a14.2±0.55b
组氨酸6.0±0.43c3.2±0.31d18.5±0.76b21.9±0.68a20.1±0.22a
赖氨酸2.3±0.09e3.2±0.31d9.9±0.28b10.8±0.08a9.5±0.20c
精氨酸3.8±0.24d6.9±0.33c24.9±0.52b29.8±0.50a29.2±0.51a
苏氨酸1.4±0.04d2.2±0.12c4.6±0.54a4.3±0.05a3.4±0.64b
色氨酸0.6±0.65c0.8±0.06c1.9±0.02a1.8±0.05a1.5±0.10b
脯氨酸1.7±0.16e4.4±0.38d22.3±0.80c51.3±0.43a35.5±0.10b
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萌芽亚麻籽中α-亚麻酸的富集工艺优化及品质分析
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何荣军 1, * , 杨雨 1 , 张锦豪 1 , 李秀双 2 , 刘芯如 1 , 王威 2
食品工业科技 | 工艺技术 2026,47(2): 277-286
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食品工业科技 |工艺技术 2026 , 47 (2) : 277 -286
萌芽亚麻籽中α-亚麻酸的富集工艺优化及品质分析
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何荣军1, * , 杨雨1, 张锦豪1, 李秀双2, 刘芯如1, 王威2
作者信息
  • 1浙江工业大学食品科学与工程学院,浙江杭州 310014
  • 2湖州维义生物科技股份有限公司,浙江湖州 313200
通讯作者:
何荣军(1978−),男,硕士,副教授,研究方向:功能性食品开发,E-mail:。
作者简介:

何荣军(1978−),男,硕士,副教授,研究方向:功能性食品开发,E-mail:

Optimization of α-Linolenic Acid Enrichment Process and Quality Analysis in Germinated Flaxseeds
Rongjun HE1, * , Yu YANG1, Jinhao ZHANG1, Xiushuang LI2, Xinru LIU1, Wei WANG2
Affiliations
  • 1College of Food Science and Engineering, Zhejiang University of Technology, Hangzhou 310014, China
  • 2Huzhou Weiyi Biotechnology Co., Ltd., Huzhou 313200, China
出版时间: 2026-01-15 doi: 10.13386/j.issn1002-0306.2025030090
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为开发富含α-亚麻酸(α-Linolenic Acid,ALA)的功能性萌芽食品,本研究以亚麻籽为原料,探究培养温度、盐浓度和pH对萌芽过程中ALA含量的影响。基于单因素实验结果,采用Box-Behnken响应面法优化培养条件,并对萌芽亚麻籽的品质及生物活性进行综合评价。结果表明,最佳萌芽时间为48 h,优化工艺参数为培养温度26 ℃、盐浓度0.5 mg/mL NaCl和pH6.0,ALA含量为20.42 g/100 g。在此条件下,脂氧合酶(Lipoxygenase,LOX)、丙二烯氧化物合成酶(Allene oxide synthase,AOS)和丙二烯氧化物环化酶(Allene oxide cyclase,AOC)的活性显著提高(P<0.05)。同时,fad2、fad3、fad7和fad8基因的表达显著上调(P<0.05),而lox基因的表达显著下调(P<0.05),从而促进了ALA的合成与积累;在最佳萌芽工艺条件下培养不同萌芽时间亚麻籽,发现与未萌芽的相比,萌芽的亚麻籽具有良好的营养价值,ALA含量达24.125 g/100 g(P<0.05),增加了13.125 g/100 g,蛋白质含量提升至25.75%(P<0.05),木酚素含量提升1.29 mg/mL(P<0.05),γ-氨基丁酸(γ-Aminobutyric acid,GABA)含量提升了28.99%(P<0.05),生氰糖苷含量降低97.62%(P<0.05);同时,生物活性显著增强(P<0.05),超氧化物歧化酶(Superoxide dismutase,SOD)活性提高154.25 U/mL(P<0.05);抗氧化能力显著改善,DPPH和ABTS+自由基清除率分别提高了12.51%和41.67%(P<0.05),铁离子还原能力达0.88 mmol/L(P<0.05)。综上所述,通过优化萌芽条件显著提升了亚麻籽中ALA的生物合成与积累,同时改善了其营养品质和生物活性,为富含ALA的健康食品研发提供了理论依据和技术支持。

α-亚麻酸  /  亚麻籽  /  萌芽  /  工艺优化  /  生物活性

In order to develop functional sprouted foods rich in α-linolenic acid (ALA), this study investigated the effects of incubation temperature, salt concentration, and pH on the ALA content during the sprouting process using flaxseed as the raw material. Based on the results of single-factorial experiments, Box-Behnken response surface methodology was used to optimize the sprouting conditions, and the quality and bioactivity of sprouted flaxseed were comprehensively evaluated. The results indicated that the optimal germination time was 48 h, and the optimized process parameters were an incubation temperature of 26 ℃, a salt concentration of 0.5 mg/mL NaCl, and a pH of 6.0, resulting in an ALA content of 20.42 g/100 g. Under these conditions, the activities of lipoxygenase (LOX), allene oxide synthase (AOS), and allene oxide cyclase (AOC) were significantly enhanced (P<0.05). Meanwhile, the expression of fad2, fad3, fad7, and fad8 genes was significantly upregulated (P<0.05), while the expression of lox gene was downregulated (P<0.05), thereby promoting the synthesis and accumulation of ALA. When flax seeds were sprouted for different periods of time under the optimal sprouting conditions, it was found that the sprouted flax seeds had better nutritional value compared with the unsprouted ones, with the ALA content reaching 24.125 g/100 g (P<0.05), an increase of 13.125 g/100 g. The protein content was increased to 25.75% (P<0.05), the lignan content was increased by 1.29 mg/mL (P<0.05), and the γ-aminobutyric acid (GABA) content was increased by 28.99% (P<0.05), while the cyanogenic glycoside content was reduced by 97.62% (P<0.05). In addition, bioactivity was significantly enhanced (P<0.05), with superoxide dismutase (SOD) activity increasing by 154.25 U/mL (P<0.05). The antioxidant capacity was also significantly improved, with the DPPH and ABTS+ free radical scavenging rates increasing by 12.51% and 41.67%, respectively (P<0.05), and the ferrous ion reducing power reaching 0.88 mmol/L (P<0.05). In conclusion, the biosynthesis and accumulation of ALA in flaxseed were significantly improved by optimized germination conditions, and its nutritional quality and bioactivity were enhanced, providing theoretical basis and technical support for the development of healthy foods rich in ALA.

α-linolenic acid  /  flaxseed  /  germination  /  process optimization  /  bioactivity
何荣军, 杨雨, 张锦豪, 李秀双, 刘芯如, 王威. 萌芽亚麻籽中α-亚麻酸的富集工艺优化及品质分析. 食品工业科技, 2026 , 47 (2) : 277 -286 . DOI: 10.13386/j.issn1002-0306.2025030090
Rongjun HE, Yu YANG, Jinhao ZHANG, Xiushuang LI, Xinru LIU, Wei WANG. Optimization of α-Linolenic Acid Enrichment Process and Quality Analysis in Germinated Flaxseeds[J]. Science and Technology of Food Industry, 2026 , 47 (2) : 277 -286 . DOI: 10.13386/j.issn1002-0306.2025030090
亚麻(Linum usitatissimum L),又称胡麻,是一种重要的油料作物和功能性食品原料,其种子因富含ω-3脂肪酸、木酚素和膳食纤维等多种生物活性成分而备受关注[1−3]。现有研究表明,亚麻籽在心血管保护、抗氧化和肠道菌群调节等方面具有潜在功效[4−5],但其应用受到生氰糖苷等抗营养因子的限制[6]。传统加工方法难以同时实现营养成分富集和抗营养因子降解的双重目标,这成为制约亚麻籽功能性开发的关键瓶颈。
α-亚麻酸(α-Linolenic Acid,ALA)是一种关键的ω-3多不饱和脂肪酸,在维持人体健康方面发挥着多种生理功能,包括但不限于代谢综合征改善[7]、抗炎作用[8]、抗氧化效应[9]以及心血管和神经系统保护[10]。亚麻籽作为植物界最丰富的ALA来源之一,其ALA含量可达脂肪酸总量的50%~60%。然而,亚麻籽中的ALA主要以甘油三酯形式存在,存在生物利用度受限的问题,且其含量易受品种、生长环境及加工工艺等因素的影响[11]。当前研究现状表明,虽然萌芽技术作为一种绿色生物加工方法已在功能性食品领域获得广泛应用,并在谷物类作物的营养改良方面取得显著成效,但针对油料作物特别是亚麻籽中ALA代谢调控的研究仍存在重要空白,如现有ALA富集研究多集中于物理化学提取工艺的优化,对生物加工途径的调控机制探索不足;关于萌芽时间对ALA合成关键酶基因表达的动态影响也尚未明确。因此,探索萌芽对ALA含量的影响及其调控机制,对于开发高ALA含量的功能性亚麻籽产品具有重要意义。
本研究以亚麻籽为研究对象,系统考察培养温度、盐浓度和pH对萌芽过程中ALA含量的影响,筛选并优化最佳萌芽工艺参数,并通过分析萌芽前后ALA代谢相关酶活性、关键基因表达差异、营养成分及生物活性的动态变化,揭示萌芽对ALA合成与积累的调控机制。研究结果旨在为高ALA含量功能性亚麻籽食品的开发提供理论依据和技术支持。
亚麻籽 会宁维义农业有限公司;石油醚、异辛烷、盐酸、氢氧化钠、硫酸氢钠、硫酸、三氯乙酸、无水乙醇 均为分析纯,上海国药集团化学试剂有限公司;甲醇、乙腈 均为色谱级,上海泰坦科技股份有限公司;抗坏血酸 色谱纯,上海麦克林生化科技股份有限公司;脂氧合酶活性测定试剂盒 伊势久生物科技有限责任公司;植物丙二烯氧化物环化酶酶联免疫分析试剂盒、植物丙二烯氧化物合成酶酶联免疫分析试剂盒 上海雅吉生物科技有限公司;FastPure® Universal Plant Total RNA lsolation Kit试剂盒、HiScript® II Q RT SuperMix for qPCR(+g DNA wiper)、ChamQ Universal SYBR qPCR Master Mix 南京诺唯赞生物科技股份有限公司;超氧化物歧化酶分型测定试剂盒、总抗氧化能力检测试剂盒(FRAP法)、ABTS+自由基清除能力试剂盒 南京建成生物工程研究所。
DHP9082恒温培养箱 中国一恒科学仪器有限公司;LC-2050高效液相色谱仪 日本岛津公司;SOX606脂肪测定仪 中国海能未来技术集团股份有限公司;UDK139半自动凯氏定氮仪 意大利VELP公司;RE3000A旋转蒸发仪 中国坤诚科学仪器有限公司;NANODROP核酸蛋白分析仪 美国赛默飞世尔科技有限公司;AnalytikJena荧光定量PCR仪 德国耶拿分析仪器有限公司;T25高速剪切机 德国 IKA公司;7820A气相色谱仪 美国安捷伦科技有限公司;CR21N落地式高速冷冻离心机 日本日立集团;FC酶标仪 美国赛默飞世尔科技有限公司;HH-6数显恒温水浴锅 常州澳华仪器有限公司;ME1D4E电子天平 瑞士梅特勒-托利多集团有限公司。
称取80 g亚麻籽,按1:7的料液比加入无菌水,清洗后浸泡30 min。随后对亚麻籽进行脱胶处理,直至表面无粘液,重复3~5次,完成后将其置于恒温培养箱中萌芽,持续4天。每24 h收集一次样品。培养温度、盐浓度和pH根据实验设计进行设置,萌芽结束后进行样品处理和指标测定。
在固定条件:料液比1:7,浸泡时间为30 min,脱胶处理的基础上,以48 h培养后的ALA含量为指标,考察培养温度(15、20、25、30、35 ℃,固定盐浓度0.7 mg/mL,pH5.0)、盐浓度(0.1、0.3、0.5、0.7、1.0 mg/mL)、pH(3.0、4.0、5.0、6.0、7.0)三个因素对ALA含量的影响,且后续因素的实验均在前一因素实验的较优结果上进行。
根据单因素实验结果,选取对ALA含量影响显著的三个因素:培养温度(A)、盐浓度(B)和pH(C),以ALA含量为响应值,进行响应面试验设计。采用Box-Behnken设计进行工艺优化试验,各因素及其水平编码见表1。
ALA的测定依照GB 5009.168-2016《食品安全国家标准 食品中脂肪酸的测定》第一法进行,经甲酯化处理后采用气相色谱法测定,其含量按公式(1)计算。
$ \mathrm{A}\mathrm{L}\mathrm{A}\mathrm{含}\mathrm{量}\left(\mathrm{g}/100\;\mathrm{g}\right)=\left(\frac{{\mathrm{A}}_{\mathrm{A}\mathrm{L}\mathrm{A}}}{\sum {\mathrm{A}}_{\mathrm{总}\mathrm{脂}\mathrm{肪}\mathrm{酸}}}\right)\times \mathrm{总}\mathrm{脂}\mathrm{肪}\mathrm{含}\mathrm{量} $
式中,AALA表示测得的ALA峰面积;∑A总脂肪酸表示所有可检测脂肪酸的峰面积总和。
采用试剂盒法测定脂氧合酶(LOX)、丙二烯氧化物合成酶(AOS)及丙二烯氧化物环化酶的活性。分别称取未萌芽亚麻籽和萌芽亚麻籽样品约2.0 g(精确至0.0001 g),使用研磨机充分研磨后,加入预冷的20 mL磷酸盐缓冲液(PBS,0.05 mol/L)进行溶解。随后,将混合物置于旋涡振荡器中充分混匀,并采用超声波破碎仪在冰浴条件下处理5~10 min。处理后的混合物于4 ℃条件下以12000 r/min离心5 min,收集上清液用于酶活性测定。酶活性的测定严格参照试剂盒说明书进行操作。
采用FastPure® Universal Plant Total RNA Isolation Kit 试剂盒提取总RNA,三个ALA含量梯度组和一个对照组:低ALA组(20 ℃、0.5 mg/mL盐浓度、pH5.0,ALA含量11.71 g/100 g)、中ALA组(25 ℃、0.7 mg/mL盐浓度、pH7.0,ALA含量14.57 g/100 g)、高ALA组(25 ℃、0.5 mg/mL盐浓度、pH6.0,ALA含量17.90 g/100 g)以及对照组(25 ℃蒸馏水培养),梯度组处理均基于响应面优化试验设计。
使用RNase-free ddH2O作为空白对照,取1 μL RNA溶液,使用核酸蛋白测定仪检测其在260~280 nm波长下的吸光度值,并计算OD260/OD280比值。若比值处于1.95~2.05范围内时,表明RNA纯度符合后续实验标准。
RNA浓度测定后,立即在冰浴条件下使用HiScript Ⅱ Q RT SuperMix for qPCR(+gDNA wiper)试剂盒进行cDNA合成,所有操作步骤均严格遵循试剂盒说明书。
根据参考文献[12]进行引物设计,所用引物见表2,采用ChamQ Universal SYBY qPCR Master Mix试剂盒进行反应,反应体系为10 µL,循环40次。
亚麻籽按照前述1.2.1方法处理,按最佳培养工艺进行萌芽。分别取未萌芽亚麻籽和萌芽亚麻籽样品,利用研磨机进行研磨,然后取适量研磨样品开展营养成分的测定。水分、蛋白质和脂肪的测定分别依据GB 5009.3-2016《食品安全国家标准 食品中水分的测定》第一法、GB 5009.5-2016《食品安全国家标准 食品中蛋白质的测定》第一法和GB 5009.6-2016《食品安全国家标准 食品中脂肪的测定》的第一法进行。
采用氨基酸自动分析仪进行样品测定。准确称取1 g样品,加入10 mL蒸馏水,振荡提取24 h。提取完成后,将混合物离心分离,收集上清液。取1 mL上清液,加入等体积的10%三氯乙酸溶液(2倍浓度),于4 ℃条件下静置过夜以沉淀蛋白质。随后,在4 ℃条件下以37920 r/min离心15 min,收集0.2 mL上清液,转移至旋转蒸发仪中蒸干。蒸干后的固形物用1 mL 5倍浓度的柠檬酸钠缓冲液溶解,并通过0.22 µm滤膜过滤,所得滤液用于上样分析。
脂肪酸的测定依据GB 5009.168-2016《食品安全国家标准 食品中脂肪酸的测定》第一法进行。
采用高效液相色谱法对样品进行测定。精确称取0.5 g样品,加入10 mL 95%乙醇和10 mL 1 mol/L NaOH溶液,于60 ℃条件下超声提取1 h。提取完成后,将混合物与4 ℃条件下以12649 r/min离心5 min,收集沉淀并用10 mL 95%乙醇洗涤,再次以12649 r/min。离心5 min。合并两次离心所得上清液,用2 mol/L H2SO4调节pH至3.0。随后,将混合物与己烷按1:5(w/v)比例混合进行脱脂处理,加蒸馏水定容至10 mL,最后通过0.45 μm有机相微孔滤膜过滤,待测定。色谱条件:色谱柱采用反相C18(5 μm,250×4.6 mm);柱温:25 ℃;进样量:10 μL;流动相:A-0.01 mol/L磷酸盐缓冲液pH=2.8,梯度洗脱程序如下:B-乙腈;A-B:0~20 min(95:5,v/v),21~22 min(80:20,v/v),23~25 min(30:70,v/v),26~30 min(95:5,v/v);流速:1 mL/min;检测波长:280 nm。以浓度为横坐标,峰面积为纵坐标绘制标准曲线,其标准方程为Y=1795.2X+2755.5(R2=0.9992)。基于该方程,计算样品中木酚素的含量。
γ-氨基丁酸的测定依据NY/T 2890-2016《稻米中γ-氨基丁酸的测定 高效液相色谱法》中高效液相色谱法进行。色谱条件:流动相流速0.3 mL/min、柱温箱温度30 ℃,流动相A为0.1%甲酸水溶液,流动相B为含0.1%甲酸的乙腈溶液。
参考文献[13]方法并稍作修改。精确称取200 mg不同萌发时间的亚麻籽样品,加入2 mL甲醇,超声提取30 min。提取结束后,将混合液在4 ℃下以15811 r/min离心10 min,收集上清液。重复上述提取和离心操作3次,合并全部上清液。取1 mL合并后的提取液,加入等体积蒸馏水混匀,经0.45 µm有机相微孔滤膜过滤,所得滤液用于后续分析。色谱条件:流动相流速0.3 mL/min、柱温箱温度30 ℃,流动相A为0.1%甲酸水溶液,流动相B为含0.1%甲酸的乙腈溶液,梯度洗脱程序如下:0~4 min,15% B;5~12 min,15%~50% B;12~12.1 min,50%~15% B;12.1~15 min,15% B。以浓度为横坐标,峰面积为纵坐标作出标准曲线,得到标准方程为Y=646.3X+9326.9(R2=0.9921)。根据标准曲线方程计算样品中生氰糖苷的含量。
称取2.0 g样品,加入50 mmol/L磷酸盐缓冲液(pH7.8,含1 mmol/L EDTA和1% PVP),按照1:10料液比充分混合。将混合液置于冰浴中,先进行机械匀浆处理,随后超声提取15 min。提取完成后,将混合液在4 ℃条件下以15000 r/min离心15 min,小心收集上清液备用。样品的超氧化物歧化酶(SOD)活性测定严格按照试剂盒说明书进行操作。
亚麻籽的体外抗氧化活性以清除DPPH自由基的能力为衡量标准,测定方法参照Noreen等[14]描述的DPPH自由基清除能力的测定方法。同时以抗坏血酸为对照组,比较两者的自由基清除能力。DPPH自由基清除能力的计算如公式(2):
$ \mathrm{D}\mathrm{P}\mathrm{P}\mathrm{H}\mathrm{自}\mathrm{由}\mathrm{基}\mathrm{清}\mathrm{除}\mathrm{率}(\text{%})=\frac{{\mathrm{A}}_{0}-{\mathrm{A}}_{1}}{{\mathrm{A}}_{0}}\times 100 $
式中,A0表示DPPH溶液的吸光度;A1表示被测样品存在时的吸光度。
称取约2 g亚麻籽样品,加入20 mL 70%甲醇,充分振荡混匀,将混合物置于超声仪中,在室温下超声提取30 min,将提取后的混合物在4 ℃条件下,以15000 r/min离心10 min,使固液分离。小心收集上清液。ABTS+自由基清除能力的测定严格参照试剂盒操作规范进行,其清除率按公式(3)计算。总抗氧化能力的测定同样依据试剂盒标准操作规程执行,最终结果以FeSO4标准溶液当量浓度(mmol/L)表示。
$ {\mathrm{A}\mathrm{B}\mathrm{T}\mathrm{S}}^+\mathrm{自}\mathrm{由}\mathrm{基}\mathrm{清}\mathrm{除}\mathrm{率}(\text{%})=\left(1-\frac{{\mathrm{A}}_{1}-{\mathrm{A}}_{2}}{{\mathrm{A}}_{0}}\right)\times 100 $
式中,A0表示蒸馏水和ABTS溶液反应后的吸光度;A1表示亚麻籽溶液与ABTS溶液反应后的吸光度;A2表示蒸馏水和亚麻籽溶液混合后的吸光度。
所有实验均重复三次,数据以均数±标准差表示,使用SPSS 27.0、Design Expert 13和GraphPad Prism 10进行统计分析和作图。多组数据采用单因素方差分析或双因素方差分析,以Tukey’s检验进行多重比较确定统计学意义,P<0.05表示有统计学意义。
温度、盐浓度和pH是调控亚麻籽萌芽及ALA生物合成的关键环境因子[15]。目前,利用萌芽处理提升亚麻籽ALA含量的研究仍较为有限。由结果可知,ALA含量随培养温度的升高呈现先上升后下降的趋势,且在25 ℃时达到峰值(图1A)。这一温度条件下,种子的呼吸代谢和酶活性处于最适状态,为ALA的高效合成与积累提供了有利的生理环境。盐浓度对ALA含量的影响表现为典型的“低促高抑”效应(图1B),在0.1~0.5 mg/mL NaCl范围内,适度的盐胁迫通过增强细胞渗透调节能力、激活抗氧化防御系统以及促进脂质代谢关键酶活性,显著(P<0.05)提高了ALA的积累;然而,当盐浓度超过0.5 mg/mL时,过量的盐胁迫抑制了合成酶活性并加剧了氧化降解,导致ALA含量显著下降。此外,pH对ALA合成的影响同样显著(图1C),ALA含量在pH5.0的弱酸性条件下ALA积累量最高,这可能与弱酸性环境能够有效促进脂肪分解和脂氧合酶途径的激活,从而优化ALA的生物合成有关[16]。基于研究结果可以推测,25 ℃培养温度、0.5 mg/mL NaCl盐浓度和pH5.0的培养条件能够在诱导亚麻籽产生适度胁迫响应的同时,有效调控脂肪酸代谢相关酶和基因的表达水平,从而最大限度地促进α-亚麻酸的生物合成与积累。
基于单因素实验结果,采用Box-Behnken试验设计原理,运用3因素3水平的响应面分析法对亚麻籽萌发条件进行优化。实验设计共包含17组试验,其中5组为中心点试验,用于评估试验误差。以培养温度(A)、盐浓度(B)和pH(C)为自变量,以ALA含量为响应值,开展响应面优化实验,具体试验结果见表3。
根据表3中的数据,采用Design-Expert 13统计分析软件进行数据分析,建立了培养温度(A)、盐浓度(B)和pH(C)的二次多项式模型,并对模型中的各项系数进行了方差分析。拟合得到的二次多项式方程为:Y=17.59+0.8138A+0.2988B+0.9325C−0.1225AB+0.2050AC+0.0650BC−1.43A2−1.33B2−2.98C2。由表4可知,模型整体极显著(P<0.0001),且失拟项不显著(P=0.9842),表明实验误差较小,模型可靠性较高。
盐浓度的一次项达到显著水平,而培养温度的一次项、pH的一次项、培养温度的二次项、盐浓度的二次项以及pH的二次项均达到极显著水平。Adeq precision值为21.2566,表明模型具有较高的预测精度。R2adj值为0.9872,说明该模型能够解释98.72%的响应值变化。综上所述,该模型拟合效果良好,适用于亚麻籽萌芽工艺的分析与预测。
采用Design-Expert 13软件绘制培养温度(A)、盐浓度(B)和pH(C)两因素之间的三维曲面图,结果如图2所示。由图2结合表4可知,两因素交互作用中培养温度与pH(AC)交互作用对ALA含量的影响最大,培养温度与盐浓度(AB)影响次之。综上,两因素对应的曲线中对萌芽亚麻籽ALA含量影响由大到小依次为培养温度与pH(AC)>培养温度与盐浓度(AB)>盐浓度与pH(BC)。根据F值大小可知影响萌芽亚麻籽ALA含量大小的因素主次顺序为:pH>培养温度>盐浓度。
经Design-Expert 13软件分析结果,亚麻籽萌芽的最佳参考工艺条件为培养温度26.463 ℃,盐浓度0.521 mg/mL,pH6.167,在此条件下萌芽亚麻籽ALA含量理论值为21.81 g/100 g。根据实际操作的可行性,调整工艺为培养温度26 ℃,盐浓度0.5 mg/mL,pH6.0,在此条件下进行3次平行验证实验,萌芽亚麻籽中ALA含量平均值为20.42 g/100 g,与预测值较为接近,说明该回归模型对于优化亚麻籽萌芽工艺是有效的。因此,应用此工艺条件进行后续实验。
在ALA代谢途径中,LOX、AOS和AOC作为核心催化酶,协同调控ALA的生物合成过程。为探究环境因子对酶活性的影响,比较了温度、盐浓度和pH变化对亚麻籽萌发过程中LOX、AOS和AOC酶活性的动态变化(图3)。结果表明,三种酶的活性均呈现先升高后降低的趋势,且均在26 ℃、0.5 mg/mL盐浓度和pH6.0附近达到峰值。这一变化趋势与ALA含量的最适条件高度一致,表明适宜的培养环境能够显著增强ALA代谢相关酶的活性。酶活性分析进一步证实,温度、盐浓度和pH是调控酶活性的关键环境因子,适宜条件能够维持酶蛋白的稳定性并提高其催化效率,而极端条件则可能导致酶构象改变、活性中心失活或底物结合受阻[17]。因此,优化培养温度、盐浓度和pH条件对于维持ALA代谢相关酶的稳定性、提高其催化效率以及促进ALA的生物合成具有重要意义。
比较了正常培养条件下与响应面优化过程中低、中、高ALA含量组萌芽亚麻籽中ALA代谢相关基因的表达差异。实验结果表明,温度、盐浓度和pH的变化显著(P<0.05)调控了ALA合成与代谢相关基因的表达水平(图4)。与对照组相比,高ALA含量组中脂肪酸去饱和酶基因fad2、fad3、fad7和fad8的mRNA相对表达量显著上调(P<0.05),其中fad2的表达量达到对照组的2.13倍,表明其高表达为ALA的生物合成提供了充足的底物[18]。同时,fad3、fad7和fad8作为催化亚油酸转化为ALA的关键基因,在高ALA含量组中的整体表达水平显著高于对照组(P<0.05)。相比之下,参与ALA氧化代谢的脂氧合酶基因lox在高ALA含量组中的表达量显著降低(P<0.05),这与lox在种子发育过程中逐渐下调的表达模式一致。lox作为催化ALA转化为脂肪酸氢过氧化物的关键酶,其高表达会加速ALA的氧化降解[19],这解释了对照组中ALA含量(约为高ALA含量组的2.45倍)显著高于高ALA含量组的原因(P<0.05)。综上所述,这5个基因共同参与调控亚麻籽萌芽过程中ALA的形成,通过上调fad2、fad3、fad7和fad8的表达,同时抑制lox的表达,从而促进油酸向亚油酸及ALA的转化,最终提高萌芽过程中ALA的积累。
未萌芽及萌芽亚麻籽的理化指标测定结果如图5所示。萌芽过程中,含水量呈现先快后慢的变化趋势,这可能与渗透压调节及细胞壁膨胀有关。蛋白质含量在萌芽48 h达到峰值(25.75%),较未萌芽状态提高7.77%,这可能与蛋白酶激活导致的储藏蛋白分解及新蛋白合成有关[20]。脂肪含量在萌芽24 h后降至最低值(33.79 g/100 g),较未萌芽时减少约15.57 g/100 g,随后回升,这一动态变化主要由于脂肪分解酶、脂酶及脂肪氧化酶的激活,促使甘油三酯水解产生游离脂肪酸和甘油,同时引发不饱和脂肪酸的氧化降解等代谢过程。
亚麻籽萌芽过程中氨基酸的动态变化是蛋白质代谢的重要表征,也是评估其营养价值提升的关键指标。如表5所示,萌芽后18种氨基酸的总含量较未萌芽状态显著增加,其中丝氨酸、谷氨酸、亮氨酸、苯丙氨酸、组氨酸、精氨酸和脯氨酸的含量变化尤为显著。这些氨基酸的积累为蛋白质的生物合成提供了必需前体物质,有效促进了细胞的增殖与修复。从营养学角度来看,氨基酸含量的增加不仅优化了亚麻籽的蛋白质品质,还赋予其多种生理功能特性:包括免疫调节、神经功能改善、心血管保护、组织修复增强以及抗氧化能力提升等[20]。
亚麻籽萌芽前后脂肪酸含量的变化如图6所示。萌芽处理显著促进了ALA的积累,在48 h时ALA达到峰值(24.125 g/100 g),较初始值(10.200 g/100 g)增加了13.925 g/100 g。与此同时,棕榈酸、油酸及亚油酸含量在48 h后呈下降趋势。这种脂肪酸组成的转变可能源于硬脂酸、油酸和亚油酸通过氧化脱氢反应向ALA的转化过程[21]。48 h后脂肪酸含量的不规则波动反映了萌芽后期脂肪酸代谢(合成、氧化及能量利用途径)的复杂动态平衡。这种代谢途径的调整不仅显著提升了ALA的相对含量,还可能增强萌芽产物的生物活性。
亚麻籽中的木酚素作为一类具有多重生物活性的天然化合物,其显著的抗氧化、抗癌、心血管保护、激素调节及肠道健康改善等生理功能,使亚麻籽成为具有重要开发潜力的功能性食品原料[22]。如图7所示,亚麻籽萌芽过程中木酚素含量呈现显著的动态变化特征。随着萌芽时间的延长,木酚素含量呈现先升高后降低的趋势,其中在萌芽48 h时达到最大值(2.56 mg/mL),较未萌芽状态显著增加了1.29 mg/mL(P<0.05)。这一动态变化规律表明了木酚素生物合成的代谢调控机制,在萌芽初期,苯丙烷代谢途径被显著激活,关键限速酶(包括苯丙氨酸解氨酶PAL、肉桂酸-4-羟化酶C4H和4-香豆酸-CoA连接酶4CL)的活性上调(P<0.05),通过松脂醇-落叶松脂醇途径促进木酚素的生物合成;而在萌芽后期,由于苯丙烷类前体物质供应不足及酶活性降低(P<0.05),木酚素合成速率下降,同时木酚素通过氧化聚合作用转化为木质素类衍生物,最终表现为含量先升高后降低的趋势[23]。
如图8所示,亚麻籽在萌芽过程中GABA含量发生了显著变化(P<0.05)。随着萌芽时间的延长,亚麻籽中的GABA含量逐渐增加,萌芽后的含量较未萌芽时提高了28.99%。尽管亚麻籽中GABA的初始含量较低,但通过萌芽或加工处理,其含量得到了显著提升。这可能与萌芽过程中谷氨酸在谷氨酸脱羧酶催化下转化为GABA的代谢过程有关,从而促进了GABA的合成与积累。此外,GABA含量的增加有助于提升亚麻籽的抗氧化能力,这不仅增强了其营养价值,也改善了其生理功能[24]。
生氰糖苷作为亚麻籽中的主要抗营养因子,主要分布于种壳和种仁中,可通过水煮、烘烤、微波加热或溶剂提取等加工方式有效去除[25]。如图9所示,亚麻籽在萌芽过程中生氰糖苷含量呈现显著下降趋势(P<0.05),且与萌芽时间呈显著负相关(P<0.05)。其中,萌芽初期的前3 d表现出快速降解特征,至48 h时生氰糖苷含量已降低97.62%。这可能是萌芽过程中内源酶的激活,其通过酶促反应加速了生氰糖苷的降解代谢,从而显著降低了该抗营养因子的含量。
亚麻籽中的超氧化物歧化酶(SOD)在抗氧化防御系统中起关键作用,其通过高效催化超氧化物转化为氧气和过氧化氢,显著提升抗氧化能力[26]。如图10所示,SOD酶活性随萌芽时间延长而增加,在48 h达到峰值(154.25 U/mL),这归因于萌芽过程中代谢活性的增强促进了抗氧化酶的合成(图10A)。同时,亚麻籽的抗氧化活性显著提升,DPPH自由基清除率从86.45%增至98.96%,ABTS+自由基清除率从55.97%增至97.64%,铁离子还原能力从0.25 mmol/L提高至1.13 mmol/L(图10B~图10D)。亚麻籽萌芽提高其抗氧化能力,可能与其代谢途径的激活密切相关,可能与萌芽激活其代谢途径,促进木酚素等天然抗氧化成分合成,增强水合作用和酶活性有关,这一过程促进了木酚素等天然抗氧化成分的生物合成,同时通过增强水合作用和提高酶活性,进一步强化了其抗氧化防御系统[27]。
本文探究了亚麻籽萌芽过程中ALA的富集机制及其品质变化规律。通过响应面法优化获得最佳萌芽工艺参数为:培养温度26 ℃、盐浓度0.5 mg/mL、pH6.0、萌芽时间48 h,在此条件下ALA含量达20.42 g/100 g。实验结果表明,该优化条件通过协同调控LOX、AOS和AOC的活性,同时上调fad2、fad3、fad7和fad8基因的表达水平,显著促进了ALA的生物合成与积累。在最佳萌芽工艺条件下,亚麻籽的营养成分组成和生物活性呈现出显著改善。与现有研究相比,本研究阐明了萌芽过程中脂肪酸代谢的分子调控机制,证实萌芽处理可同步提升亚麻籽的营养价值和功能活性。然而,研究也发现萌芽后期ALA含量出现下降趋势,其分子调控机制及不同品种亚麻籽对萌芽处理的响应差异仍需进一步探究。基于此,建议未来研究应结合转录组学和代谢组学技术,深入解析代谢调控机制,并进一步探索规模化生产工艺的优化方案。本研究为开发富含ALA的功能性产品研发提供新的思路,同时也为亚麻籽等油料作物精深加工和产业化应用提供了重要的理论依据和实践参考。

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2026年第47卷第2期
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doi: 10.13386/j.issn1002-0306.2025030090
  • 接收时间:2025-03-10
  • 首发时间:2026-10-24
  • 出版时间:2026-01-15
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  • 收稿日期:2025-03-10
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    1浙江工业大学食品科学与工程学院,浙江杭州 310014
    2湖州维义生物科技股份有限公司,浙江湖州 313200

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何荣军(1978−),男,硕士,副教授,研究方向:功能性食品开发,E-mail:。
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2种不同金属材料的力学参数
科
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
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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