Article(id=1320889157195510272, tenantId=1146029695717560320, journalId=1260987677001138203, issueId=1320888908896907909, articleNumber=null, orderNo=null, doi=10.13386/j.issn1002-0306.2025020126, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1739980800000, receivedDateStr=2025-02-20, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1792854116008, onlineDateStr=2026-10-24, pubDate=1772294400000, pubDateStr=2026-03-01, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792854116008, onlineIssueDateStr=2026-10-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792854116008, creator=13701087609, updateTime=1792854116008, updator=13701087609, issue=Issue{id=1320888908896907909, tenantId=1146029695717560320, journalId=1260987677001138203, year='2026', volume='47', issue='5', pageStart='1', pageEnd='503', issueExtLink='null', onlineDate='null', pubDate='1772294400000', pubDateStr='2026-03-01', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792854056809, creator='13701087609', updateTime=1792854056809, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=186, endPage=197, ext={EN=ArticleExt(id=1320889157422002689, articleId=1320889157195510272, tenantId=1146029695717560320, journalId=1260987677001138203, language=EN, title=Screening, Identification, and Characterization of Aroma-producing Lactic Acid Bacteria in Mustard Fermentation, columnId=1261336275769016441, journalTitle=Science and Technology of Food Industry, columnName=Bioengineering, runingTitle=null, highlight=null, articleAbstract=

To obtain lactic acid bacteria (LAB) with aroma-producing capabilities for application in mustard fermentation, traditional isolation and culturing methods were employed to screen presumptive LAB strains from fermented vegetables. Aroma-producing LAB were selected using sniffing test and diacetyl content analysis. Growth and fermentation characteristics of the strains were studied, and they were inoculated into mustard juice to analyze volatile aroma components. The results showed that: A total of 67 LAB strains were isolated, from which 5 aroma-producing strains were screened. Based on morphological, physiological, biochemical, and 16S rDNA molecular identification, strains LJ036, LJ065, QT038, and JC091 were identified as Lactiplantibacillus plantarum, while JC045 was identified as Weissella hellenica. LJ065 and QT038 exhibited stronger acid-producing capabilities. QT038 grew well at pH3.5, showed slightly higher salt tolerance, and demonstrated good temperature tolerance. LJ036 exhibited the strongest antibacterial effect against Staphylococcus aureus. LJ065 possessed the highest nitrite degradation ability, with a degradation rate of 94.45%. HS-SPME-GC-MS (headspace solid-phase microextraction gas chromatography-mass spectrometry) identified 52 volatile compounds. The fermented groups contained higher levels of alcohols, aldehydes, and ketones. Among them, the QT038, LJ065, and LJ036 fermentation groups showed higher acetoin contents of 4.27, 3.11, and 3.80 μg/mL, respectively. Cluster heatmap analysis revealed higher levels of volatile components such as (Z)-2-hexen-1-ol, damascenone, (Z)-3-hexen-1-ol, 2-ethylphenol, and linalyl acetate in QT038, LJ065, and LJ036 fermented mustard juice, imparting more intense floral, fruity, and herbaceous notes to the mustard greens. Comprehensive evaluation indicated that QT038, LJ065, and LJ036 possessed favorable fermentation and aroma-producing characteristics and showed promise as potential starter cultures for inoculated fermentation of mustard.

, authors=Huiting LAI1, Wei LIU1, 2, *, Yixuan LUO1, Xinyu WANG1, Shuhua LIN2, Juhua ZHANG1, 2, *, authorsList=Huiting LAI, Wei LIU, Yixuan LUO, Xinyu WANG, Shuhua LIN, Juhua ZHANG, authorCompany=null, correspAuthors=Wei LIU, Juhua ZHANG, 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=1320889160286712348, articleId=1320889157195510272, tenantId=1146029695717560320, journalId=1260987677001138203, language=CN, title=芥菜发酵用产香乳酸菌的筛选、鉴定及特性研究, columnId=1261336277547401341, journalTitle=食品工业科技, columnName=生物工程, runingTitle=null, highlight=null, articleAbstract=

为获得具有产香能力的乳酸菌应用于芥菜发酵生产,采用传统分离培养法从发酵蔬菜中筛选疑似乳酸菌,通过嗅闻法和乙偶姻含量分析筛选产香乳酸菌,开展菌株生长及发酵特性研究,并接种至芥菜汁中分析其挥发性香气成分。结果表明:本研究共获得67株乳酸菌菌株,筛到5株产香菌株,经形态学、生理生化和16S rDNA分子生物学鉴定,LJ036、LJ065、QT038和JC091为植物乳植杆菌,JC045为赫伦魏斯氏菌。LJ065和QT038具有更强的产酸能力;QT038在pH3.5下生长良好,且高盐耐受力略强,具有较好的耐温性;LJ036对金黄色葡萄球菌的抑菌效果最好;LJ065的亚硝酸盐降解能力最强,降解率为94.45%。HS-SPME-GC-MS挥发性成分共鉴定出52种,发酵组挥发性成分含量较高的有醇类、醛类和酮类,其中QT038、LJ065和LJ036发酵组的乙偶姻含量较高,分别为4.27、3.11、3.80 μg/mL。聚类热图分析显示QT038、LJ065和LJ036发酵芥菜汁中(Z)-2-己烯-1-醇、大马士酮、(Z)-3-己烯-1-醇、2-乙基苯酚和乙酸芳樟酯等挥发性成分含量较高,赋予芥菜更为浓郁的花香、果香及草木香。综合评价,QT038、LJ065和LJ036具备较好的发酵及产香特性,有望为芥菜接种发酵提供菌株支持。

, authors=赖慧婷1, 刘伟1, 2, *, 罗艺璇1, 王新宇1, 林树花2, 张菊华1, 2, *, authorsList=赖慧婷, 刘伟, 罗艺璇, 王新宇, 林树花, 张菊华, authorCompany=null, correspAuthors=刘伟, 张菊华, authorNote=

赖慧婷(2000−),女,硕士研究生,研究方向:果蔬发酵加工,E-mail:

, correspAuthorsNote=
刘伟(1983−),男,博士,研究员,研究方向:果蔬加工及综合利用,E-mail:。
张菊华(1971−),女,硕士,研究员,研究方向:果蔬发酵加工,E-mail:。
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赖慧婷(2000−),女,硕士研究生,研究方向:果蔬发酵加工,E-mail:

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赖慧婷(2000−),女,硕士研究生,研究方向:果蔬发酵加工,E-mail:

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注:不同小写字母表示有显著性差异,P<0.05。

, figureFileSmall=DxZkb/5ZrJ2BZOxO7RQV4Q==, figureFileBig=nPxsgaBOWUxBRW/SMH7tnw==, tableContent=null), ArticleFig(id=1320889164032225873, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.3, caption=Colony morphology and Gram staining result of 5 strains, figureFileSmall=0z48/m88z1P0sE0hi89lAg==, figureFileBig=wt7K3ZqeYS0kAqAj/np8rA==, tableContent=null), ArticleFig(id=1320889164099334738, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图3, caption=5株菌的形态和革兰氏染色结果, figureFileSmall=0z48/m88z1P0sE0hi89lAg==, figureFileBig=wt7K3ZqeYS0kAqAj/np8rA==, tableContent=null), ArticleFig(id=1320889164162249299, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.4, caption=Results of 16S rDNA, figureFileSmall=r0+nKvz5bmqxU6Tq5iQxCQ==, figureFileBig=ektJaUT6uA1k7juaOi7N2A==, tableContent=null), ArticleFig(id=1320889164229358164, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图4, caption=16S rDNA结果, figureFileSmall=r0+nKvz5bmqxU6Tq5iQxCQ==, figureFileBig=ektJaUT6uA1k7juaOi7N2A==, tableContent=null), ArticleFig(id=1320889164292272725, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.5, caption=Phylogenetic tree of strains, figureFileSmall=ux3xxj2xMviz/14GsmEiFQ==, figureFileBig=vwz8WhsfHt7N3kFwlxJgXA==, tableContent=null), ArticleFig(id=1320889164359381590, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图5, caption=菌株系统发育树, figureFileSmall=ux3xxj2xMviz/14GsmEiFQ==, figureFileBig=vwz8WhsfHt7N3kFwlxJgXA==, tableContent=null), ArticleFig(id=1320889164430684759, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.6, caption=Growth curve and pH of 5 strains, figureFileSmall=V2L6xT+LcpLYXr02ZonqOA==, figureFileBig=LNzObLKzx+4coeccllbw2g==, tableContent=null), ArticleFig(id=1320889164514570840, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图6, caption=5株菌的生长曲线和pH, figureFileSmall=V2L6xT+LcpLYXr02ZonqOA==, figureFileBig=LNzObLKzx+4coeccllbw2g==, tableContent=null), ArticleFig(id=1320889164585874009, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.7, caption=Growth of the strains at different pH, figureFileSmall=8/0sp1S8INCQbxAptZ4NQA==, figureFileBig=PWAD48I6bS1WbJ85z8ew9A==, tableContent=null), ArticleFig(id=1320889164652982874, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图7, caption=不同pH下各菌株的生长情况

注:同pH,不同小写字母表示有显著性差异,P<0.05。

, figureFileSmall=8/0sp1S8INCQbxAptZ4NQA==, figureFileBig=PWAD48I6bS1WbJ85z8ew9A==, tableContent=null), ArticleFig(id=1320889164715897435, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.8, caption=Inhibition zones of the strains in different indicator bacteria, figureFileSmall=HvL5gTZjkAQdCK7SI5CIYw==, figureFileBig=Up4EDqcp0dNRHpiGW8KReA==, tableContent=null), ArticleFig(id=1320889164803977820, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图8, caption=不同指示菌中各菌株的抑菌圈

注:上为金黄色葡萄球菌,下为大肠杆菌。

, figureFileSmall=HvL5gTZjkAQdCK7SI5CIYw==, figureFileBig=Up4EDqcp0dNRHpiGW8KReA==, tableContent=null), ArticleFig(id=1320889164866892381, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.9, caption=Antibacterial activity of 5 strains, figureFileSmall=CmDG4+fA2407coXPPX2fsA==, figureFileBig=1qvGm8yFoSI0mLKK2cA3UA==, tableContent=null), ArticleFig(id=1320889164934001246, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图9, caption=5株菌的抑菌能力

注:同指示菌,不同小写字母表示有显著性差异,P<0.05。

, figureFileSmall=CmDG4+fA2407coXPPX2fsA==, figureFileBig=1qvGm8yFoSI0mLKK2cA3UA==, tableContent=null), ArticleFig(id=1320889165026275935, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.10, caption=Color reaction results of strains with hydrochloric acid 1-naphthylamine, figureFileSmall=tbC5vxKEIqNSwTp6JCS1XA==, figureFileBig=yrG5Tpag8XnYT7dpx5tJiw==, tableContent=null), ArticleFig(id=1320889165101773408, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图10, caption=菌株的盐酸萘乙二胺显色反应效果图, figureFileSmall=tbC5vxKEIqNSwTp6JCS1XA==, figureFileBig=yrG5Tpag8XnYT7dpx5tJiw==, tableContent=null), ArticleFig(id=1320889165206631009, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.11, caption=Antioxidant capacity determination results of lactic acid bacteria strains, figureFileSmall=jPZMuiFTXVQGzcpNgLE8Bg==, figureFileBig=y/jftAQtaR7tolya+9B5Ww==, tableContent=null), ArticleFig(id=1320889165282128482, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图11, caption=乳酸菌菌株抗氧化能力的测定结果

注:同指标,不同小写字母表示有显著性差异,P<0.05。

, figureFileSmall=jPZMuiFTXVQGzcpNgLE8Bg==, figureFileBig=y/jftAQtaR7tolya+9B5Ww==, tableContent=null), ArticleFig(id=1320889165340848739, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.12, caption=Types (A) and relative content (B) in volatile components in different fermented mustard, figureFileSmall=Rtt+HHjd205rl3u9VNPTrg==, figureFileBig=lEgXoPwGP6kxZ7XRy1aZZQ==, tableContent=null), ArticleFig(id=1320889165407957604, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图12, caption=不同发酵组芥菜挥发性成分种类数(A)和含量(B), figureFileSmall=Rtt+HHjd205rl3u9VNPTrg==, figureFileBig=lEgXoPwGP6kxZ7XRy1aZZQ==, tableContent=null), ArticleFig(id=1320889165462483557, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Fig.13, caption=Cluster heat map analysis of volatile aroma compounds in mustard juice fermented by lactic acid bacteria, figureFileSmall=fWURkA+b81WExBEMz96xHA==, figureFileBig=J/JIKeNE61fzxLjr5g5g3A==, tableContent=null), ArticleFig(id=1320889165542175334, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=图13, caption=乳酸菌发酵芥菜汁的挥发性香气成分聚类热图分析, figureFileSmall=fWURkA+b81WExBEMz96xHA==, figureFileBig=J/JIKeNE61fzxLjr5g5g3A==, tableContent=null), ArticleFig(id=1320889165609284199, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Table 1, caption=

Physiological and biochemical results

, figureFileSmall=null, figureFileBig=null, tableContent=
序号项目LJ036LJ065QT038JC091JC045序号项目LJ036LJ065QT038JC091JC045
1阴性对照−−−−−26七叶灵+++++
2丙三醇−−−−−27水杨苷+++++
3赤藓糖醇−−−−−28D-纤维二糖+++++
4D-阿拉伯糖−−−−−29D-麦芽糖+++++
5L-阿拉伯糖+++++30D-乳糖+++++
6D-核糖+++++31D-密二糖++++−
7D-木糖−−−−+32D-蔗糖+++++
8L-木糖−−−−−33D-海藻糖+++++
9D-核糖醇−−−−−34菊粉−−−−−
10甲基-β-D吡喃木糖苷−−−−−35D-松三糖++++−
11D-半乳糖+++++36D-棉子糖−−−−−
12D-葡萄糖+++++37淀粉−−−−−
13D-果糖+++++38糖原−−−−−
14D-甘露糖+++++39木糖醇−−−−−
15L-山梨糖−−−−−40D-龙胆二糖+++++
16L-鼠李糖++++−41D-土伦糖++++−
17卫矛醇−−−−−42D-来苏糖−−−−−
18肌醇−−−−−43D-塔格糖−−−−−
19甘露醇+++++44D-岩藻糖−−−−−
20山梨醇++++−45L-岩藻糖−−−−−
21甲基-α-D-吡喃甘露糖苷++++−46D-阿拉伯醇−−−−−
22甲基-α-D-吡喃葡萄糖苷−−−−−47L-阿拉伯醇−−−−−
23N-乙酰葡萄糖胺+++++48葡萄糖酸钾++++−
24苦杏仁苷+++++492-酮基葡萄糖酸钾−−−−−
25熊果苷+++++505-酮基葡萄糖酸钾−−−−−
), ArticleFig(id=1320889165693170280, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=表1, caption=

生理生化结果

, figureFileSmall=null, figureFileBig=null, tableContent=
序号项目LJ036LJ065QT038JC091JC045序号项目LJ036LJ065QT038JC091JC045
1阴性对照−−−−−26七叶灵+++++
2丙三醇−−−−−27水杨苷+++++
3赤藓糖醇−−−−−28D-纤维二糖+++++
4D-阿拉伯糖−−−−−29D-麦芽糖+++++
5L-阿拉伯糖+++++30D-乳糖+++++
6D-核糖+++++31D-密二糖++++−
7D-木糖−−−−+32D-蔗糖+++++
8L-木糖−−−−−33D-海藻糖+++++
9D-核糖醇−−−−−34菊粉−−−−−
10甲基-β-D吡喃木糖苷−−−−−35D-松三糖++++−
11D-半乳糖+++++36D-棉子糖−−−−−
12D-葡萄糖+++++37淀粉−−−−−
13D-果糖+++++38糖原−−−−−
14D-甘露糖+++++39木糖醇−−−−−
15L-山梨糖−−−−−40D-龙胆二糖+++++
16L-鼠李糖++++−41D-土伦糖++++−
17卫矛醇−−−−−42D-来苏糖−−−−−
18肌醇−−−−−43D-塔格糖−−−−−
19甘露醇+++++44D-岩藻糖−−−−−
20山梨醇++++−45L-岩藻糖−−−−−
21甲基-α-D-吡喃甘露糖苷++++−46D-阿拉伯醇−−−−−
22甲基-α-D-吡喃葡萄糖苷−−−−−47L-阿拉伯醇−−−−−
23N-乙酰葡萄糖胺+++++48葡萄糖酸钾++++−
24苦杏仁苷+++++492-酮基葡萄糖酸钾−−−−−
25熊果苷+++++505-酮基葡萄糖酸钾−−−−−
), ArticleFig(id=1320889165789639273, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Table 2, caption=

Growth of lactic acid bacteria strains in different concentrations of NaCl

, figureFileSmall=null, figureFileBig=null, tableContent=
食盐浓度(g/L)菌株名称
LJ036LJ065QT038JC091JC045
601.988±0.029a1.850±0.076b1.987±0.011a1.780±0.021c1.721±0.019c
801.100±0.058d1.381±0.017a1.370±0.080ab1.251±0.013bc1.245±0.029c
1000.193±0.016c0.256±0.017a0.235±0.007ab0.227±0.008ab0.224±0.017abc
1200.169±0.016a0.182±0.008a0.183±0.007a0.182±0.006a0.179±0.013a
1400.152±0.004b0.168±0.008a0.175±0.009a0.171±0.007a0.165±0.004ab
), ArticleFig(id=1320889165848359530, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=表2, caption=

乳酸菌在不同NaCl浓度下生长情况

, figureFileSmall=null, figureFileBig=null, tableContent=
食盐浓度(g/L)菌株名称
LJ036LJ065QT038JC091JC045
601.988±0.029a1.850±0.076b1.987±0.011a1.780±0.021c1.721±0.019c
801.100±0.058d1.381±0.017a1.370±0.080ab1.251±0.013bc1.245±0.029c
1000.193±0.016c0.256±0.017a0.235±0.007ab0.227±0.008ab0.224±0.017abc
1200.169±0.016a0.182±0.008a0.183±0.007a0.182±0.006a0.179±0.013a
1400.152±0.004b0.168±0.008a0.175±0.009a0.171±0.007a0.165±0.004ab
), ArticleFig(id=1320889165915468395, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Table 3, caption=

Growth of the strains at different temperatures

, figureFileSmall=null, figureFileBig=null, tableContent=
温度(℃)菌株名称
LJ036LJ065QT038JC091JC045
40.245±0.001c0.262±0.001c0.351±0ab0.368±0.003a0.315±0.001b
372.218±0.020a2.189±0.075a2.205±0.018a2.251±0.072a2.188±0.052a
451.821±0.087ab1.888±0.060a1.880±0.053a1.760±0.022b1.839±0.006ab
), ArticleFig(id=1320889165965800044, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=表3, caption=

不同温度下各菌株的生长情况

, figureFileSmall=null, figureFileBig=null, tableContent=
温度(℃)菌株名称
LJ036LJ065QT038JC091JC045
40.245±0.001c0.262±0.001c0.351±0ab0.368±0.003a0.315±0.001b
372.218±0.020a2.189±0.075a2.205±0.018a2.251±0.072a2.188±0.052a
451.821±0.087ab1.888±0.060a1.880±0.053a1.760±0.022b1.839±0.006ab
), ArticleFig(id=1320889166045491821, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=EN, label=Table 4, caption=

NaNO2 reduction rate of different strains

, figureFileSmall=null, figureFileBig=null, tableContent=
菌株名称LJ036LJ065QT038JC091JC045
降解率(%)93.11±0.17b94.45±0.13a91.58±0.40c91.51±0.13c85.09±0.55d
), ArticleFig(id=1320889166116794990, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320889157195510272, language=CN, label=表4, caption=

不同菌株的亚硝酸盐降解率

, figureFileSmall=null, figureFileBig=null, tableContent=
菌株名称LJ036LJ065QT038JC091JC045
降解率(%)93.11±0.17b94.45±0.13a91.58±0.40c91.51±0.13c85.09±0.55d
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芥菜发酵用产香乳酸菌的筛选、鉴定及特性研究
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赖慧婷 1 , 刘伟 1, 2, * , 罗艺璇 1 , 王新宇 1 , 林树花 2 , 张菊华 1, 2, *
食品工业科技 | 生物工程 2026,47(5): 186-197
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食品工业科技 |生物工程 2026 , 47 (5) : 186 -197
芥菜发酵用产香乳酸菌的筛选、鉴定及特性研究
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赖慧婷1 , 刘伟1, 2, * , 罗艺璇1, 王新宇1, 林树花2, 张菊华1, 2, *
作者信息
  • 1湖南大学生物学院隆平分院,湖南长沙 410125
  • 2湖南省农产品加工与质量安全研究所,湖南长沙 410125
通讯作者:
刘伟(1983−),男,博士,研究员,研究方向:果蔬加工及综合利用,E-mail:。
张菊华(1971−),女,硕士,研究员,研究方向:果蔬发酵加工,E-mail:。
作者简介:

赖慧婷(2000−),女,硕士研究生,研究方向:果蔬发酵加工,E-mail:

Screening, Identification, and Characterization of Aroma-producing Lactic Acid Bacteria in Mustard Fermentation
Huiting LAI1 , Wei LIU1, 2, * , Yixuan LUO1, Xinyu WANG1, Shuhua LIN2, Juhua ZHANG1, 2, *
Affiliations
  • 1Longping Branch, College of Biology, Hunan University, Changsha 410125, China
  • 2Hunan Institute of Agricultural Product Processing and Quality Safety, Changsha 410125, China
出版时间: 2026-03-01 doi: 10.13386/j.issn1002-0306.2025020126
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为获得具有产香能力的乳酸菌应用于芥菜发酵生产,采用传统分离培养法从发酵蔬菜中筛选疑似乳酸菌,通过嗅闻法和乙偶姻含量分析筛选产香乳酸菌,开展菌株生长及发酵特性研究,并接种至芥菜汁中分析其挥发性香气成分。结果表明:本研究共获得67株乳酸菌菌株,筛到5株产香菌株,经形态学、生理生化和16S rDNA分子生物学鉴定,LJ036、LJ065、QT038和JC091为植物乳植杆菌,JC045为赫伦魏斯氏菌。LJ065和QT038具有更强的产酸能力;QT038在pH3.5下生长良好,且高盐耐受力略强,具有较好的耐温性;LJ036对金黄色葡萄球菌的抑菌效果最好;LJ065的亚硝酸盐降解能力最强,降解率为94.45%。HS-SPME-GC-MS挥发性成分共鉴定出52种,发酵组挥发性成分含量较高的有醇类、醛类和酮类,其中QT038、LJ065和LJ036发酵组的乙偶姻含量较高,分别为4.27、3.11、3.80 μg/mL。聚类热图分析显示QT038、LJ065和LJ036发酵芥菜汁中(Z)-2-己烯-1-醇、大马士酮、(Z)-3-己烯-1-醇、2-乙基苯酚和乙酸芳樟酯等挥发性成分含量较高,赋予芥菜更为浓郁的花香、果香及草木香。综合评价,QT038、LJ065和LJ036具备较好的发酵及产香特性,有望为芥菜接种发酵提供菌株支持。

芥菜  /  乳酸菌  /  乙偶姻  /  菌株鉴定  /  生长特性

To obtain lactic acid bacteria (LAB) with aroma-producing capabilities for application in mustard fermentation, traditional isolation and culturing methods were employed to screen presumptive LAB strains from fermented vegetables. Aroma-producing LAB were selected using sniffing test and diacetyl content analysis. Growth and fermentation characteristics of the strains were studied, and they were inoculated into mustard juice to analyze volatile aroma components. The results showed that: A total of 67 LAB strains were isolated, from which 5 aroma-producing strains were screened. Based on morphological, physiological, biochemical, and 16S rDNA molecular identification, strains LJ036, LJ065, QT038, and JC091 were identified as Lactiplantibacillus plantarum, while JC045 was identified as Weissella hellenica. LJ065 and QT038 exhibited stronger acid-producing capabilities. QT038 grew well at pH3.5, showed slightly higher salt tolerance, and demonstrated good temperature tolerance. LJ036 exhibited the strongest antibacterial effect against Staphylococcus aureus. LJ065 possessed the highest nitrite degradation ability, with a degradation rate of 94.45%. HS-SPME-GC-MS (headspace solid-phase microextraction gas chromatography-mass spectrometry) identified 52 volatile compounds. The fermented groups contained higher levels of alcohols, aldehydes, and ketones. Among them, the QT038, LJ065, and LJ036 fermentation groups showed higher acetoin contents of 4.27, 3.11, and 3.80 μg/mL, respectively. Cluster heatmap analysis revealed higher levels of volatile components such as (Z)-2-hexen-1-ol, damascenone, (Z)-3-hexen-1-ol, 2-ethylphenol, and linalyl acetate in QT038, LJ065, and LJ036 fermented mustard juice, imparting more intense floral, fruity, and herbaceous notes to the mustard greens. Comprehensive evaluation indicated that QT038, LJ065, and LJ036 possessed favorable fermentation and aroma-producing characteristics and showed promise as potential starter cultures for inoculated fermentation of mustard.

mustard greens  /  lactic acid bacteria  /  acetoin  /  strain identification  /  growth characteristics
赖慧婷, 刘伟, 罗艺璇, 王新宇, 林树花, 张菊华. 芥菜发酵用产香乳酸菌的筛选、鉴定及特性研究. 食品工业科技, 2026 , 47 (5) : 186 -197 . DOI: 10.13386/j.issn1002-0306.2025020126
Huiting LAI, Wei LIU, Yixuan LUO, Xinyu WANG, Shuhua LIN, Juhua ZHANG. Screening, Identification, and Characterization of Aroma-producing Lactic Acid Bacteria in Mustard Fermentation[J]. Science and Technology of Food Industry, 2026 , 47 (5) : 186 -197 . DOI: 10.13386/j.issn1002-0306.2025020126
芥菜(Brassica juncea)属于十字花科植物,是一年生草本植物[1]。芥菜营养丰富,含有大量的膳食纤维、叶绿素、β-胡萝卜素、抗坏血酸、矿物质和挥发性成分等[2−3],同时富含生物活性成分,如硫代葡萄糖苷及其降解产物和多酚(类黄酮和花青素)。发酵芥菜因其口感脆嫩和风味鲜美,具有多种健康益处和疾病预防作用[4−5],受到消费者的喜爱。
发酵蔬菜的风味源于微生物群的代谢活动、菌群之间以及与非生物环境(原料、盐、温度)的相互作用,非生物环境通过影响微生物活性、群落结构及相互作用来影响风味[6−7]。自然发酵蔬菜大多依赖于原料中自带的乳酸菌(Lactic acid bacteria)进行发酵,如肠膜明串珠菌(Leuconostoc mesenteroides)、粪肠球菌(Enterococcus faecalis)、乳酸乳球菌(Lactococcus lactis)以及植物乳植杆菌(Lactiplantibacillus plantarum)等[8]。华容大叶芥菜在不同的盐浓度下发酵,其主要的细菌种类包括乳杆菌属(Lactobacillus)、片球菌属(Pediococcus)和棒状杆菌属(Corynebacterium)等[9]。不动杆菌属(Acinetobacter)、普兰久浩酵母属(Guehomyces)、假单胞菌属(Pseudomonas)和魏斯氏菌属(Weissella)则是泡青菜发酵过程中风味形成的关键微生物[10]。但目前传统芥菜发酵大多采用高盐保坯工艺,发酵风味不足,同时由于周期长,风味品质不稳定,特定的植物乳植杆菌对芥菜的品质和代谢产物产生显著的影响,筛选适宜的菌株可进一步丰富腌制芥菜的风味多样性[11]。
发酵芥菜中主要的风味成分包括醇类、醛类、酯类、酸类、酮类以及含硫和含氮的化合物,其中乙醛、乙偶姻及双乙酰等是乳酸菌发酵腌制品的主要芳香来源[12]。而乙醛在发酵过程中容易被进一步代谢为其它物质,如乙醇;双乙酰的生成途径较为复杂且在某种情况下可能会产生不良风味,这两种物质作为筛选指标都不够理想[13]。乙偶姻(Acetoin,3-hydroxy-2-butanone)具有令人愉快的牛奶味、奶油味、甜香味[14],可由二乙酰基和2,3-丁二醇转化合成、丁酮水解或醛直接合成,也可以由微生物产生(例如,在奶酪和酒精发酵过程中乳酸菌、酵母代谢生成乙偶姻) [15−16],在发酵过程中具有较高的稳定性和积累性,容易检测和监控,因此作为筛选产香乳酸菌指标更为合适。有研究从香椿中筛选产香菌株,该菌株发酵后乙偶姻含量显著提升至714.6 mg/kg[17];乙偶姻随腌菜腌渍时间的延长,含量增加[18]。因此,发酵芥菜行业急需挖掘具有优良风味特性的发酵菌株,如分离高产乙偶姻的乳酸菌菌株制备优良发酵剂并应用于蔬菜发酵,从而提升发酵芥菜的品质风味。
本研究从发酵蔬菜中分离乳酸菌,采用感官嗅闻和产乙偶姻能力(肌酸比色法)综合考量,获得产香优良的菌株,再经形态学、生理生化和16S rDNA分子生物学鉴定,同时探究乳酸菌的生长曲线、耐盐、耐酸、耐温、抑菌、降解亚硝酸盐特性及抗氧化特性,分析其发酵芥菜汁的产香特性,获得产香和生长特性优良的乳酸菌。
新鲜大叶芥菜 湖南洞庭明珠食品有限公司提供;发酵蔬菜及泡菜汁 收集于湖南省华容县、新宁县、保靖县等78份;大肠杆菌、金黄色葡萄球菌 湖南省农产品加工与质量安全研究所实验室保存;葡萄糖(D(+)-葡萄糖,一水)、氯化钠、对氨基苯磺酸、铁氰化钾、乙酸锌、肌酸(一水)、亚硝酸钠、磷酸氢二钾、硼砂 国药集团化学试剂有限公司;N-(1-萘基)乙二胺盐酸盐 天津市化学试剂研究所有限公司;酵母浸粉 广东环凯微生物科学研究所;丙三醇 西陇化工股份有限公司;环己酮、甲醇 色谱纯,德国Merck公司;1-萘酚 北京沃凯生物科技有限公司;邻苯二胺 麦克林化学试剂有限公司;0.1 mol/L盐酸标准溶液 深圳市博林达科技有限公司;乙偶姻 标准品,上海阿拉丁生物化学科技有限公司;革兰氏染色试剂盒、VP试剂、30%过氧化氢 广东环凯微生物科学研究所;DPPH法试剂盒、ABTS法试剂盒、FRAP试剂盒 苏州科铭生物技术有限公司;Ezup柱式细菌基因组DNA抽提试剂盒 生工生物工程(上海)股份有限公司;MRS肉汤培养基、LB琼脂培养基 广东环凯微生物科学研究所;改良MRS琼脂培养基 广州华润科技有限公司;MRS-NaNO2培养基:每1 L水中加入54 g的MRS肉汤,加热搅拌溶解,pH调至6.8,再加入5 mg NaNO2,含NaNO2浓度为200 mg/L。
PHS-3C型pH计 上海仪电科学仪器股份有限公司;WP-UP-WF-20型超纯水制备机 四川沃特尔水处理设备有限公司;SW-CJ-1B型超净工作台 苏州净化设备有限公司;LX-B100 L型立式压力蒸汽灭菌锅 合肥华泰有限公司;SPX-150型生化培养箱 中仪国科(北京)科技有限公司;Synergy H1型酶标仪 美国Bio-Tek公司;Avanti J-26XP冷冻离心机 美国Beckman公司;50/30 μm固相萃取仪 德国CNW公司;7890A-5975C型气相色谱-质谱(Gas chromatography-mass spectrometry,GC-MS)联用仪 美国 Agilent公司。
发酵蔬菜或泡菜汁分别取1 g(mL)于10 mL MRS肉汤培养基,37 ℃培养1 d,培养液经梯度稀释后,从10−3、10−4、10−5、10−6稀释梯度中分别吸取0.1 mL涂布在MRS固体培养基上,37 ℃培养2~3 d。用接种环选取单菌落进行平板划线分离,纯化2~3次。
菌株活化后,进行革兰氏染色和过氧化氢酶试验,选取疑似乳酸菌、无芽孢、革兰氏阳性和过氧化氢接触酶阴性的菌株保存。
发酵芥菜汁制作工艺流程如图1所示。参考张佳等[19]感官评定标准,选6名经过培训具有感官评定背景的人员,评定方法如下:以未接种芥菜汁为对照(CK),采用嗅闻法对不同的发酵芥菜汁打分。评分标准:0~3分:无明显发酵香气或有异味;4~7分:有正常发酵香气;8~10分:有浓郁发酵香气(每个样品评3次)。同时用pH计测定pH。
采用肌酸比色法[20]测定发酵芥菜汁中的乙偶姻。准确称取2.5 g乙偶姻,定容至50 mL容量瓶中,配制成质量浓度为50 mg/mL的乙偶姻标准溶液,并稀释至不同倍数。以乙偶姻质量浓度(X)为横坐标,530 nm处吸光值(Y)为纵坐标绘制标准曲线,获得标准曲线回归方程Y=0.0186X−0.0052,R2=0.9990。
取5 mL发酵液用蒸馏水定容至50 mL,混匀后静置30 min,取上清液于8000 r/min离心10 min,取离心后的上清液3 mL,加入1 mL的5% 1-萘酚,1 mL的0.5% 肌酸,振荡2 min,37 ℃下反应60 min,红色为阳性反应,未变色则为阴性反应。以不接菌种作为空白对照,测定反应液在530 nm处吸光值(A),利用标准曲线回归方程计算乙偶姻含量。
将获得的高产乙偶姻菌株和嗅闻香气好综合评价优的菌株接于MRS固体培养基中,37 ℃下培养48 h后观察并拍照记录菌株形态特征,经革兰氏染色后置于显微镜油镜下观察并记录。
碳源利用能力采用法国梅里埃公司API 50 CHL试纸条和API 50 CH对筛选优产乙偶姻菌株进行鉴定,检测菌株对49种不同碳源的发酵能力。按照API试纸条使用手册将活化后的菌株接种到试纸中,用无菌液体石蜡封口,记录培养24 h和48 h的结果,利用API LAB Plus自动判读系统鉴定。
采用CTAB法提取菌株DNA。16S rDNA扩增采用通用引物。测序引物为:27F:AGAGTTTGATCMTGGCTCAG;1492R:GGTTACCTTGTTACGACTT。PCR反应体系(共25 μL):10×PCR Buffer,dNTP(each 10 mmol/L),Taq Plus DNA Polymerase(5 U/μL)和50 mmol/L MgSO4共12.5 μL;引物F(10 µmol/L)1 μL;引物R(10 µmol/L)1 μL;Template(DNA)1 μL;ddH2O 9.5 μL。PCR反应体系为:95 ℃预变性5 min;94 ℃变性30 s;57 ℃退火30 s;72 ℃延伸90 s,共循环30次;最后72 ℃延伸10 min。将筛选出的菌株经2~3次纯化出单菌落送至生工生物工程(上海)股份有限公司测序,将测序结果与NCBI中GenBank数据库进行BLAST对比,采用MEGA 7.0构建系统发育树。
将活化后的菌株以2%接种量接种于MRS肉汤培养基中37 ℃培养24 h,以MRS培养基为空白,每2 h用酶标仪测定600 nm处吸光值(A)以及测定pH,每组3平行。
将活化好的菌株以2%接种量分别接至含60、80、100、120、140 g/L NaCl的MRS液体培养基中,37 ℃培养24 h,测定600 nm处吸光值(A),每组3平行。
将活化好的菌株以2%接种量分别接至pH2.0、2.5、3.0、3.5、4.0的MRS液体培养基中,37 ℃培养24 h,测定600 nm处吸光值(A),每组3平行。
将活化好的菌株以2%接种量分别接至MRS液体培养基中,于4、37、45 ℃培养24 h,测定600 nm处吸光值(A),每组3平行。
选取大肠杆菌(Escherichia coil)、金黄色葡萄球菌(Staphylococcus aureus)作为指示菌。采用琼脂扩散法[21]检测乳酸菌的抑菌能力,测量抑菌圈直径并按公式计算抑菌能力(Antimicrobial Unit,AU)。
$ \mathrm{AU={\pi} (d}_{ \mathrm{1}}^{ \mathrm{2}} \mathrm{-d}_{ \mathrm{0}}^{ \mathrm{2}} \mathrm{)/4V} $
式中:AU:抑菌能力,mm2/mL;d1:抑菌圈直径,mm;d0:琼脂孔径,7.5 mm;V:抑菌液体积,0.2 mL。
参照郭志华等[22]的方法,以不接种含亚硝酸盐的MRS培养基为空白对照,并测定反应液538 nm处吸光值(A)。按公式计算菌株的亚硝酸盐降解率:
$\rm 亚硝酸盐降解率(\text{%})=\frac{ {\mathrm{C}}_{0}-{\mathrm{C}}_{1}}{{\mathrm{C}}_{0}}\times 100 $
式中:C0和C1分别为培养基中NaNO2的含量和接种菌株发酵后NaNO2的含量。
待测样品:将各菌株接入MRS肉汤培养基中,37 ℃培养24 h后,于4 ℃、5000 r/min离心10 min,取上清液待测。
采用总抗氧化活性(DPPH)试剂盒(微量法)进行测定。空白:试剂盒中提取液20 µL与380 µL DPPH溶液混合。测定:取上清液20 µL与380 µL DPPH溶液充分混匀。室温避光反应20 min,取200 µL测定515 nm处吸光值(A)。按公式计算DPPH自由基的清除率:
$\rm DPPH自由基清除率(\text{%})=\frac{ {\mathrm{A}}_{\text{空白}}-{\mathrm{A}}_{\text{测定}}}{{\mathrm{A}}_{\text{空白}}}\times 100 $
采用总抗氧化活性(ABTS)试剂盒(微量法)进行测定。空白:试剂盒中提取液10 µL与190 µL ABTS溶液混合。测定:取上清液10 µL与190 µL ABTS溶液充分混匀。需10 min内测定734 nm处吸光值(A)。按公式计算ABTS+自由基的清除率:
$\rm ABTS^+自由基清除率(\text{%})= \frac{{\mathrm{A}}_{\text{空白}}-{\mathrm{A}}_{\text{测定}}}{{\mathrm{A}}_{\text{空白}}}\times 100 $
混合液制备(现配现用):由0.3 mol/L醋酸缓冲盐、10 mmol/L 2,4,6-三(2-吡啶基)-1,3,5-三嗪和20 mmol/L FeCl3溶液按10:1:1的比例混合,使用前37 ℃预温。
采用总抗氧化活性(FRAP)试剂盒(微量法)进行测定。空白:试剂盒中提取液10 µL和190 µL 混合液混匀。测定:取上清液10 µL与190 µL 混合液充分混匀。反应20 min,测定593 nm处吸光值(A)。还原力以Trolox为标准,以mmol Trolox/L表示,按公式计算样品对铁离子的还原能力:
$ {\rm FRAP还原力(mmol\; Trolox/L)=\rm (A_{测定}-A_{空白}-0.0134)\times 0.8054 }$
采用SPME-GC-MS[23]对发酵芥菜汁的香气成分进行富集测定。取5 mL芥菜汁至20 mL顶空萃取瓶中,随后加入0.85 g NaCl和10 µL内标物(环己酮浓度为18.94 μg/mL),于40 ℃、500 r/min 平衡30 min。萃取头于250 ℃老化1 h,萃取头在40 ℃下萃取1 h,随后取出并插入GC仪进样口,250 ℃解析5 min后检测。采用DB-5MS毛细管柱(30 m×0.25 mm,0.25 µm)分离挥发性化合物;载气为氦气(1.2 mL/min),程序升温:40 ℃以3 ℃/min速度升至100 ℃,再以5 ℃/min升至200 ℃,最后以10 ℃/min升至260 ℃;质谱采用70 eV电子电离,离子源和传输线温度分别为230 ℃和280 ℃;检测采用扫描方式,检测距离为50~550 m/z。利用GC-MS分析软件分析,通过比对NIST 2011质谱库匹配度、CAS号、各峰值保留时间对挥发性香气化合物进行定性,采用内标物定量,计算各发酵芥菜汁中香气成分的含量。
$ \rm 香气成分含量 (\text{μ}g/mL)= \frac{香气峰面积}{内标的峰面积}\times 内标的浓度$
所有实验均进行3次重复,结果以平均数±标准差表示。采用IBM SPSS 25.0和Origin Pro 2024软件进行方差分析和作图。P<0.05表示为差异有统计学意义。
发酵芥菜、辣椒等发酵蔬菜经梯度稀释后,于MRS固体培养基培养,共筛得疑似乳酸菌88株。对筛得的88株疑似乳酸菌菌株进行革兰氏染色和过氧化氢接触酶实验,获得革兰氏染色呈阳性且接触酶阴性的67株菌。
根据消费者感官偏好的重要性,香气浓郁、典型乳酸发酵香味是重要的评价指标[24]。通过嗅闻法得到有浓郁发酵香气菌株25株,有正常发酵香气菌株34株,无明显发酵香气或有异味菌株8株;产酸能力强(pH≤3.50)有5株,分别是LJ036、LJ065、LJ071、DJ001和LJ033;pH3.50~3.60的有15株;pH3.60~3.70的最多,有23株。结果得到优良菌株40株,均具有浓郁发酵香气或正常发酵香气,且发酵芥菜汁后pH小于3.70。
采用肌酸比色法,在碱性条件下,发酵液与肌酸混合液变成红色则表明该菌株具有产乙偶姻的能力[25]。显色反应结果见图2(A),在40株乳酸菌中有15株菌株呈现粉色至深红色。
将肌酸比色法得到显色深的菌株进行乙偶姻含量的测定。图2(B)所示为15株菌株的乙偶姻测定值,其中乙偶姻含量较高的有QT038、LJ036、LJ065、JC045和JC091,含量分别为57.50、43.29、36.03、17.68和11.42 mg/L,均高于10 mg/L。综合考量乳酸发酵香气好、pH低以及高乙偶姻含量的菌株,选择LJ036、LJ065、QT038、JC091和JC045这5株菌株进行后续实验研究。
菌株LJ036、LJ065、QT038、JC091和JC045的菌落形态以及光学显微镜下成像如图3所示,菌落中等大小,形态均呈圆形、表面光滑、边缘整齐,颜色呈白色,革兰氏染色皆为阳性,且于光学显微镜下菌株形态均呈杆或短杆状,推测这5株菌均属于乳杆菌属。
利用API 50 CH微生物生化鉴定系统测定菌株对49种碳源的利用能力,将对照作为阴性,底物可以利用的反应结果作为阳性,试验结果见表1,于法国梅里埃公司官网利用API LABPlus自动判读系统对其进行鉴定,初步鉴定LJ036、LJ065、QT038、JC091和JC045均为植物乳植杆菌,但仅通过生理生化还不能精确判断,因此还需要进行分子生物学鉴定。
LJ036、LJ065、QT038、JC091和JC045菌株的16S rDNA基因PCR扩增结果如图4所示,条带亮度清晰,基因片段大小为1500 bp左右,测序结果得到片段长分别为1503、1530、1529、1530、1512 bp,具有典型的16S rDNA特征,符合测序要求。
运用BLAST同源性分析基因测序结果,构建系统发育树,结果见图5,其中LJ036、LJ065、QT038和JC091菌株在同一进化分支,且与乳植杆菌属模式菌株中的植物乳植杆菌NR 025447.1同源性最高;菌株JC045与魏斯氏菌属模式菌株中的赫伦魏斯氏菌NR118771.1同源性最高。基于API系统对罕见菌或新物种的鉴定能力有限,再结合形态学及生理生化实验结果,最终鉴定LJ036、LJ065、QT038和JC091菌株为植物乳植杆菌(Lactiplantibacillus plantarum),JC045菌株鉴定为赫伦魏斯氏菌(Weissella hellenica)。
测定菌株的生长曲线,可以了解菌株生长过程中的活力变化,确定菌株较优培养时间,直观评价菌株的发酵性能[26]。菌株的生长曲线及产酸测定结果见图6,5株菌在MRS肉汤中均生长良好,0~2 h时,生长比较缓慢,处于迟缓期,2~8 h快速生长,进入对数生长期,8 h后所有乳酸菌到达稳定期,600 nm处吸光值一直保持较高水平,20~24 h进入衰亡期。其中,菌株QT038在20 h时吸光值相对较高。
菌株的产酸能力是评价发酵剂的重要指标之一,影响发酵蔬菜的风味和稳定性[27]。图6可看出菌株产酸与生长同步,在2~8 h时pH快速下降,LJ036、LJ065、QT038、JC091和JC045发酵终点的pH分别为3.62、3.60、3.60、3.66和3.64,LJ065和QT038菌株表现相对较强的产酸能力。生长和产酸速度是选择菌株作芥菜发酵剂的两个重要因素,5株菌均在2 h后进入对数生长期,同时大量产酸,能迅速占据发酵初期优势地位,保证发酵质量。
乳酸菌发酵蔬菜需加入一定食盐以增加产品的风味、脆度及贮藏性,因此乳酸菌对食盐的耐受性是评价菌株的一个重要指标[28]。不同NaCl浓度下各菌株生长情况如表2所示,5株菌株的生长随NaCl浓度升高吸光值呈下降趋势,60 g/L NaCl浓度生长几乎不受影响,600 nm处吸光值均大于1.7;80 g/L NaCl浓度下吸光值为1.100~1.381,其中LJ036 耐盐能力相对较弱;超过100 g/L NaCl浓度对菌株影响较大,5株菌生长明显受到抑制,相比之下,QT038在高盐条件下耐受力略强。5株菌均可以用于芥菜的低盐(≤80 g/L NaCl)接种发酵,如需提升菌株的高盐耐受性,还要在高盐环境中逐步驯化。
菌株耐酸存活率越高,在人体胃肠道消化系统中越能较好地发挥作用。5株乳酸菌的耐酸实验结果如图7,随着pH的降低,5株菌生长呈下降趋势。pH4.0对菌株生长影响小;pH3.5下JC091和QT038生长良好;当pH≤3.0时仍能生长,但表现出较弱的pH耐受性;发酵芥菜pH3.5~4.0,会对菌株有一定的抑制作用。结合2.4.1中产酸能力发现,产酸能力强的LJ065和QT038,耐酸能力不是最优;而耐酸能力较强的JC091的产酸能力也不是最强。因此,菌株的产酸与耐酸之间是否具有关联性还有待研究。
将各菌株接种于MRS培养基中,在不同温度下培养发酵24 h后,测得各发酵液在600 nm下的吸光值,从表3可看出,5株乳酸菌株均在37 ℃温度下生长旺盛,发酵液吸光值均在2.2左右,且5株菌株表现无显著差异;在4 ℃中表现出明显的生长抑制,QT038和JC091表现相对良好的耐低温特性;45 ℃下LJ065和QT038的高温耐受性更好。可见过高或过低的温度均会抑制乳酸菌的生长,可能影响发酵产物的形成,从而影响乳酸发酵蔬菜的品质。
乳酸菌在发酵过程中能够分泌多种抑菌物质,如有机酸、细菌素和过氧化氢等。这些物质不仅能够有效抑制杂菌的生长,还能显著延长食品的保质期。此外,直接利用具有抑菌特性的乳酸菌作为生物保护菌,应用于发酵制品和食品原料的保鲜,已成为当前食品科学研究的热点领域[29−30]。
选取了两种常见的肠道致病菌:金黄色葡萄球菌和大肠杆菌,考察4株植物乳植杆菌和1株赫伦魏斯氏菌对两种致病菌在平板上生长的抑制作用,测定其抑菌圈直径大小并计算抑菌能力。从图8可以看出,LJ036对金黄色葡萄球菌的生长呈现出了较好的抑制作用,抑菌圈直径为23.12±1.75 mm;JC091对大肠杆菌的生长呈现出了较好的抑制作用,抑菌圈直径为19.20±0.53 mm。双全等[31]从发酵酸菜中分离14株乳酸菌,8株对大肠杆菌和金黄色葡萄球菌有抑菌作用,其中S1-4抑菌效果最强,抑菌圈直径分别为19.69±0.71 mm和20.59±0.26 mm。图9可知不同菌株对两种指示菌的抑制效果存在差异,5株乳酸菌均能抑制两种指示菌,其中LJ036对金黄色葡萄球菌的抑菌效果更好,JC091对大肠杆菌的抑菌能力较强。
亚硝酸盐是酱腌菜的重要安全指标之一[32]。乳酸菌是泡菜中降解亚硝酸盐效果较理想的微生物,乳酸杆菌因产酸能力强具有比乳酸球菌更优的降解亚硝酸盐能力,如植物乳植杆菌、短促生乳杆菌、发酵粘液乳杆菌等,其中植物乳植杆菌的降解效果最好[33]。5株乳酸菌的亚硝酸盐显色结果和降解率如图10、表4所示,降解率均在85%以上,其中LJ065的亚硝酸盐降解率最高为94.45%,其次是LJ036。陈雪梅等[34]采用植物乳植杆菌发酵黄秋葵泡菜的亚硝酸盐含量低于自然发酵,自然发酵亚硝酸盐含量最高为17.58 mg/kg,接种发酵亚硝酸盐含量最高为9.78 mg/kg,且成熟期更短;杨晶[35]筛选的3株植物乳植杆菌对亚硝酸盐的降解率均达到80%以上。
乳酸菌具有清除活性氧,缓解氧化损失的作用[36],还能改善发酵食品的风味缩短发酵周期。梁小波等[37]将高抗氧化活性发酵乳杆菌接种发酵泡菜,缩短了2 d左右的发酵周期并改善了风味。
由图11可以看出,5株产香乳酸菌均具有一定的抗氧化活性,但不同菌株之间的抗氧化能力存在显著差异(P<0.05)。其中5株菌的DPPH自由基清除率均超过50%,尤其LJ065菌株表现出更强的抗氧化活性,其清除率高达57.16%,显著高于赵鑫等[38]从传统泡菜中筛选菌株的50.62%。相较其它菌株,LJ065、JC091和LJ036的ABTS+自由基清除率较高,分别为53.72%、49.52%和47.57%。FRAP还原力在0.6880~1.0054 mmol Trolox/L之间,其中LJ036还原力最高为1.0054 mmol Trolox/L,其次是JC045和JC091,分别为0.8292 mmol Trolox/L和0.8086 mmol Trolox/L,与卢佳音[39]从发酵酸菜中筛选得到乳酸菌抗氧化能力结果相当。综上所述,植物乳植杆菌和赫伦魏斯氏菌具有一定的抗氧化活性,且植物乳植杆菌LJ036与LJ065的总抗氧化活性高于其他菌株。
乳酸菌发酵过程中代谢糖、蛋白质及其它营养物质,生成挥发性风味物质和非挥发性滋味物质,从而形成发酵蔬菜的特征风味[40]。风味是发酵蔬菜制品给人的最直观感受,芥菜原有的“生味”和“冲味”会被独特浓郁的酸香味代替[41]。图12(A)显示,发酵芥菜汁共鉴定出52种挥发性成分,其中醇类13种、醛类4种、酸类4种、酯类3种、酚类6种、烷类3种、烯类3种、酮类7种、杂环类7种和腈类2种。CK中挥发性成分26种,接种发酵后均显著增加了挥发性成分的种类,其中植物乳植杆菌LJ065种类最多,达42种。从图12(B)可以看出,乳酸菌接种发酵后的芥菜汁中各类挥发性风味化合物含量均有提高,且不同发酵组挥发性香气成分有差异。各组的醇类、醛类和酮类含量较高,如(Z)-4-己烯-1-醇、苯乙醇、2,4-二甲基苯甲醛、3-羟基-2-丁酮(乙偶姻)等挥发性香气物质。醇类的种类及相对含量占比均最高,醇类一般具有花香、芳香和水果香,可以与有机酸形成酯类物质,极大地改善发酵后产品的风味形成[42];发酵组的醛类化合物含量均减少,LJ036、QT038的酯类物质相对含量较高,可能是因为醛类极易被乳酸菌分解还原成醇类[43],醇类物质的增加进而会使相应的酯类物质增加,而酯类是产生浓郁水果香味的重要物质;发酵组的酮类含量均增加,酮类多伴有果香味[44],是植物乳植杆菌发酵过程中的第二类主要挥发性成分[45]。
将分析鉴定出的52种挥发性风味物质绘制成聚类热图(图13),颜色越蓝表示挥发性成分含量较高,颜色越红表示挥发性成分含量越低。由图可见CK组和5种菌株发酵组的挥发性成分有显著差异,十八烷基乙烯基醚和1-戊烯-3-醇这2种挥发物在CK组中的含量相对较高;JC091中壬酸、3-蒈烯、1-戊烯-3-醇、2-丁基辛醇和4-庚基苯酚等的含量相对较高,JC045中苯乙醇、3-蒈烯和3-丁烯腈等的含量相对较高;LJ036中4-庚基苯酚、β-紫罗酮、3,4-二甲基苯甲醛、D-柠檬烯、(Z)-2-己烯-1-醇、2-丁酮、(Z)-2-戊烯-1-醇、异戊酸和愈创木酚等的含量相对较高;LJ065中辛酸、3-甲基苄醇、2-丁酮、1-庚三醇、大马士酮、2-壬酮、芳樟醇和异戊酸等的含量相对较高;QT038中(Z)-3-己烯-1-醇、乙酸芳樟酯、2-乙基苯酚、3-羟基-2-丁酮和2,4-二叔丁基酚等的含量相对较高。苯乙醇、1-戊烯-3-醇、3-丁烯腈及(Z)-3-己烯-1-醇等是发酵芥菜、泡菜和腌制菜中主要的挥发性成分[46−48],β-紫罗酮、大马士酮、2-壬酮与芳樟醇等赋予发酵芥菜花香、果香[49−50],有效中和发酵产生的部分刺激性气味,丰富其风味层次。聚类分析可以看出JC091和JC045的风味特征较为相似,且表现为3-蒈烯含量较高、3-甲基苄醇含量居中、D-柠檬烯含量低;LJ036、LJ065和QT038的风味特征较为相似,且均表现为(Z)-2-己烯-1-醇含量较高,β-环柠檬醛含量居中,1-戊烯-3-醇含量较低。
结果表明,不同乳酸菌发酵后的芥菜汁挥发性物质存在显著差异,乳酸菌发酵可以增加挥发性香气物质的种类及含量。LJ036、LJ065、QT038、JC091、JC045菌株中均检测到3-羟基-2-丁酮(乙偶姻)含量,赋予发酵芥菜汁的甜香、奶油香。其中QT038、LJ065和LJ036发酵后芥菜汁的乙偶姻含量较高,分别为4.27、3.11和3.80 μg/mL,且其他特征挥发性成分种类较多,如(Z)-2-己烯-1-醇、大马士酮、(Z)-3-己烯-1-醇、2-乙基苯酚和乙酸芳樟酯等,这些物质香气特征为花香、果香及草木香,突显出这3株菌具有较强的风味形成能力。
本研究从发酵蔬菜或泡菜中筛选得到产酸、产香、产乙偶姻含量较高的5株乳酸菌,经形态学、生理生化和分子生物学鉴定,LJ036、LJ065、QT038和JC091为植物乳植杆菌,JC045为赫伦魏斯氏菌。其中菌株LJ065和QT038具有较强的产酸能力;5株菌低于80 g/L NaCl生长良好,QT038耐高盐能力略强;QT038表现出较好的耐低pH和耐低温;LJ065和QT038表现出较好的高温耐受性;LJ036对金黄色葡萄球菌的抑菌效果最好;LJ065菌株的亚硝酸盐降解能力最强,降解率为94.45%,其次是LJ036。研究结果表明,LJ036、LJ065和QT038具备良好的生长及发酵特性。
接种发酵芥菜汁共鉴定出52种挥发性成分,LJ065的挥发性成分种类最高达42种。各组的醇类、醛类和酮类含量较高,其中醇类的种类及相对含量占比均最高,相比CK组,发酵芥菜汁的醛类含量均减少,但酮类含量均增加,且LJ036和QT038的酯类相对含量较高。聚类热图分析表明,5株乳酸菌表现出不同的产香特性,尤其是QT038、LJ065和LJ036发酵后的芥菜汁中,(Z)-2-己烯-1-醇、大马士酮、(Z)-3-己烯-1-醇、2-乙基苯酚和乙酸芳樟酯等挥发性成分的含量相对较高,这些物质的香气特征为花香、果香及草木香,赋予芥菜更为浓郁的风味;QT038、LJ065和LJ036发酵组的乙偶姻含量较高,分别为4.27、3.11、3.80 μg/mL,突显出这3株菌具有较强的风味形成能力。本研究筛选获得的优良菌株有望为芥菜接种发酵提供有效的菌株支持。

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2026年第47卷第5期
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doi: 10.13386/j.issn1002-0306.2025020126
  • 接收时间:2025-02-20
  • 首发时间:2026-10-24
  • 出版时间:2026-03-01
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  • 收稿日期:2025-02-20
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    1湖南大学生物学院隆平分院,湖南长沙 410125
    2湖南省农产品加工与质量安全研究所,湖南长沙 410125

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刘伟(1983−),男,博士,研究员,研究方向:果蔬加工及综合利用,E-mail:。
张菊华(1971−),女,硕士,研究员,研究方向:果蔬发酵加工,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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