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江西产腐乳生物胺水平调查及产胺菌的分离与鉴定

王维亚 黄瑞钰 骆瑜 吴珊珊 刘玉婷 李金林

王维亚,黄瑞钰,骆瑜,等. 江西产腐乳生物胺水平调查及产胺菌的分离与鉴定[J]. 食品工业科技,2022,43(12):246−252. doi:  10.13386/j.issn1002-0306.2021110004
引用本文: 王维亚,黄瑞钰,骆瑜,等. 江西产腐乳生物胺水平调查及产胺菌的分离与鉴定[J]. 食品工业科技,2022,43(12):246−252. doi:  10.13386/j.issn1002-0306.2021110004
WANG Weiya, HUANG Ruiyu, LUO Yu, et al. Investigation of the Biogenic Amine Content in Jiangxi-made Sufu and Isolation and Identification of Amine-producing Bacteria[J]. Science and Technology of Food Industry, 2022, 43(12): 246−252. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2021110004
Citation: WANG Weiya, HUANG Ruiyu, LUO Yu, et al. Investigation of the Biogenic Amine Content in Jiangxi-made Sufu and Isolation and Identification of Amine-producing Bacteria[J]. Science and Technology of Food Industry, 2022, 43(12): 246−252. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2021110004

江西产腐乳生物胺水平调查及产胺菌的分离与鉴定

doi: 10.13386/j.issn1002-0306.2021110004
基金项目: 江西省市场监督管理局科技项目(GSJK202010);国家自然科学基金(32060557)。
详细信息
    作者简介:

    王维亚(1982−),女,硕士,工程师,研究方向:食品微生物,E-mail:544106094@qq.com

    通讯作者:

    李金林(1983−),男,博士,高级工程师,研究方向:食品加工,E-mail:lijinlin405@126.com

  • 中图分类号: TS201.6

Investigation of the Biogenic Amine Content in Jiangxi-made Sufu and Isolation and Identification of Amine-producing Bacteria

  • 摘要: 采用高效液相色谱法对江西产腐乳生物胺含量水平进行调查分析,通过微生物学分离结合高效液相色谱确认,筛选出生物胺产生菌,并对其进行鉴定。结果表明,12种腐乳样品中总生物胺含量范围为68.5~1084.0 mg/kg,色胺、腐胺、组胺和酪胺是主要生物胺,其中腐胺和酪胺的含量较高。筛选出的9株产胺菌产生物胺的含量范围为93.8~369.2 mg/L。对产胺量较高的6株菌进行表型鉴定和基因型16S rDNA序列测定,得到耐热芽孢杆菌(Bacillus sporothermodurans)1株,鲁梅利杆菌(Rummeliibacillus stabekisii)1株,耐硼赖氨酸芽孢杆菌(Lysinacillus boronitolerans)3株和皮脂葡萄球菌(Staphylococcus piscifermentans)1株。研究结果对腐乳食用安全性评价提供了理论依据,为进一步探清如何抑制腐乳中产胺菌的活性提供了理论研究基础。
  • 图  1  生物胺混合标准品溶液的液相色谱图

    Figure  1.  Chromatogram of biogenic amines standards

    图  2  腐乳中生物胺的液相色谱图

    Figure  2.  Chromatogram of biogenic amines in sufu

    图  3  菌株的革兰氏染色

    Figure  3.  Gram staining of strains F3、F5 and F6

    图  4  PCR产物电泳图

    Figure  4.  Agarose gel electrophoresis of PCR products

    表  1  样品信息

    Table  1.   Sample information

    编号名称产地配料
    1豆腐乳江西南昌黄豆、辣椒、植物油、味精、食用盐、白酒、白砂糖、香辛料、食品添加剂:硫酸钠、脱氢乙酸钠
    2A茶油腐乳香辣味江西吉安水、黄豆、茶油、食用植物油、食用盐、辣椒粉、香辛料
    3A农家腐乳猛辣型江西吉安水、黄豆、植物油、食用盐、辣椒粉、香辛料
    4B红油腐乳香辣味江西吉安水、大豆、菜籽油、食用盐、白酒、辣椒、八角、食品添加剂:谷氨酸钠
    5B白腐乳原味江西吉安水、大豆、食用盐、白酒、食品添加剂:脱氢乙酸钠
    6C香辣腐乳江西南昌黄豆、植物油、辣椒、食盐、酒、香辛料
    7C茶油腐乳江西南昌黄豆、菜籽精炼油、山茶油、辣椒、食盐、酒、香辛料
    8D家乡豆腐乳江西南昌黄豆、水、食用大豆油、麻油、食用碘盐、辣椒、白酒、生姜、味精、香辛料
    9D豆腐乳家乡味江西南昌黄豆、水、食用大豆油、食用碘盐、辣椒、白酒、生姜、味精、香辛料
    10山茶油腐乳江西吉安黄豆、饮用水、山茶油、辣椒、食用盐、白酒、香辛料
    11E红油腐乳王江西吉安水、大豆、菜籽油、食用盐、辣椒、白酒、香辛料、味精
    12E茶油腐乳香辣味江西吉安水、大豆、菜籽油、山茶油、食用盐、辣椒、白酒、香辛料、味精
    注:编号中的相同大写字母表示同一品牌不同系列样品;表4~表7同。
    下载: 导出CSV

    表  2  梯度洗脱程序

    Table  2.   Gradient elution procedures

    时间(min)022253232.0134
    流动相A(%)587890905858
    流动相B(%)422210104242
    下载: 导出CSV

    表  3  生物胺的保留时间、回归方程和相关系数

    Table  3.   Retention time, the regression equations and correlation coefficients of biogenic amines

    生物胺保留时间(min)回归方程相关系数
    色胺10.722Y=0.5257x+0.00160.9999
    苯乙胺13.057Y=0.6156x+0.00630.9999
    腐胺14.176Y=1.7395x+0.01070.9999
    尸胺15.569Y=1.3790x+0.00030.9999
    组胺16.369Y=0.7710x+0.01860.9990
    章鱼胺17.840Y=0.7449x+0.01580.9998
    酪胺23.545Y=1.0711x−0.01270.9998
    亚精胺25.448Y=1.4346x+0.00080.9999
    精胺30.937Y=1.3006x−0.00580.9999
    下载: 导出CSV

    表  4  腐乳样品中生物胺含量 (mg/kg)

    Table  4.   Contents of biogenic amines in sufu samples (mg/kg)

    编号色胺苯乙胺腐胺尸胺组胺章鱼胺酪胺亚精胺精胺总生物胺
    166.7±1.7g15.1±1.1ef181.8±4.7i14.6±0.8d99.8±3.7aND208.4±3.7fNDND586.8±10.0f
    2A160.2±1.3a42.3±1.7b370.2±10.4d5.7±0.7e78.0±1.2dND431.7±11.7aNDND1084.0±22.8a
    3A89.3±2.9f23.5±1.4d411.5±8.4b7.6±1.3e71.1±1.6eND414.4±4.2bNDND1013.6±8.4b
    4B152.3±2.1b35.8±3.4c326.7±9.8f5.1±0.6e82.9±2.9cND192.5±3.2gNDND797.1±20.9d
    5B110.6±4.6d13.7±1.9f353.8±9.3e18.5±1.7c85.4±2.5cND385.7±6.9cNDND967.9±11.3c
    6CNDND33.4±2.5j24.8±2.2bNDND10.3±0.9kNDND68.5±5.6i
    7CND6.4±0.9g446.4±12.1a41.4±2.5a37.5±1.7gND50.2±2.0iND12.9±1.5595.9±13.0f
    8D105.1±3.5e25.3±2.2d316.9±6.8f6.0±0.6e37.6±1.5gND343.4±9.7dNDND824.5±17.2d
    9D59.1±4.3h18.0±2.3e279.0±13.5g7.2±1.4e43.7±1.7fND328.3±8.4eNDND735.6±14.8e
    10NDND12.5±1.4kNDNDND119.4±2.3hNDND131.7±4.4h
    11E118.2±4.8c45.8±2.7a220.1±9.8h40.2±2.1a90.5±2.8bND26.7±1.9jNDND534.4±20.3g
    12E84.8±4.4f23.2±1.1d387.1±8.0c14.8±1.7d94.5±4.5bND421.7±21.8abNDND1022.5±35.7b
    注:ND表示未检出;同列的不同小写字母表示有显著性差异,P<0.05;表5同。
    下载: 导出CSV

    表  5  菌株的生物胺生成情况 (mg/L)

    Table  5.   Biogenic amine contents produced by strains (mg/L)

    编号色胺苯乙胺腐胺尸胺组胺章鱼胺酪胺亚精胺精胺总生物胺
    F1-3A3.8±0.6cND17.8±0.8dNDNDNDNDNDND21.5±0.6f
    F2-2AND80.1±2.1a66.4±2.2aNDNDND222.7±5.1aNDND369.2±8.8a
    F3-2AND28.5±0.9cNDND37.9±1.7cND177.8±2.9bNDND244.1±2.9b
    F4-5B3.3±0.6cND23.0±1.5bcNDNDNDNDNDND26.3±2.0ef
    F5-12E25.8±1.7bND20.6±1.3dND47.3±1.1bNDNDNDND93.8±1.5d
    F6-12E85.3±3.9a56.8±2.9bNDNDNDND14.6±0.7cNDND156.7±7.5c
    F7-3A6.1±0.6cND10.6±1.3eNDNDND14.1±0.9cNDND30.8±1.8e
    F8-3A23.8±1.0bND24.2±0.4bND48.0±2.0bNDNDNDND96.0±1.0d
    F9-5B23.1±2.0bND21.1±1.4cdND54.3±1.8aNDNDNDND98.5±3.4d
    注:编号中“−”后面的序号表示来源于哪种腐乳;表6~表7同。
    下载: 导出CSV

    表  6  菌株的形态学鉴定

    Table  6.   Morphological identification of strains

    编号TSA平板菌落形态革兰氏染色结果
    F2-2A乳白色,近圆形,表面平坦,有光泽革兰氏阳性芽孢杆菌
    F3-2A乳白色,近圆形,略隆起,边缘整齐革兰氏阳性杆菌
    F5-12E微黄色,不规则,表面干燥有褶皱,边缘略不整齐革兰氏阳性芽孢杆菌
    F6-12E乳白色,圆形,表面光泽湿润,边缘整齐革兰氏阳性葡萄球菌
    F8-3A微黄色,不规则,表面干燥有褶皱,边缘略不整齐革兰氏阳性芽孢杆菌
    F9-5B微黄色,不规则,表面干燥有褶皱,边缘略不整齐革兰氏阳性芽孢杆菌
    下载: 导出CSV

    表  7  16S rDNA 序列 Blast 比对

    Table  7.   Blast analysis of 16S rDNA sequences

    菌株编号鉴定结果匹配度(%)
    F2-2A耐热芽孢杆菌(Bacillus sporothermodurans97.57
    F3-2A鲁梅利杆菌(Rummeliibacillus stabekisii99.65
    F5-12E耐硼赖氨酸芽孢杆菌(Lysinacillus boronitolerans99.24
    F6-12E皮脂葡萄球菌(Staphylococcus piscifermentans99.72
    F8-3A耐硼赖氨酸芽孢杆菌(Lysinibacillus boronitolerans97.07
    F9-5B耐硼赖氨酸芽孢杆菌(Lysinibacillus boronitolerans99.24
    下载: 导出CSV
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出版历程
  • 收稿日期:  2021-11-02
  • 网络出版日期:  2022-05-14
  • 刊出日期:  2022-06-08

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