CN111424002B - 一株大洋芽孢杆菌及其应用 - Google Patents
一株大洋芽孢杆菌及其应用 Download PDFInfo
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Abstract
本发明属于微生物食品加工技术领域，尤其涉及一株大洋芽孢杆菌及其应用。本发明从襄阳大头菜卤水中筛选出了一株嗜盐菌菌株，经形态、理化和16S rRNA序列鉴定为大洋芽孢杆菌，该菌株能够显著提高腌制蔬菜中挥发性物质种类及含量，降低辛辣味物质的含量，能够改善腌制大头菜的口感风味，具有良好的开发应用前景。
Description
技术领域
本发明属于微生物食品加工技术领域，尤其涉及一株大洋芽孢杆菌及其应用。
背景技术
腌制蔬菜在我国已有很长时间的历史，有着悠久丰富的文化底蕴。腌制蔬菜含盐量高达20％，属于高盐食品。高盐食品中存在着大量的嗜盐菌，嗜盐菌在食品加工及贮藏过程中发挥着重要作用，将嗜盐菌作为食品发酵剂加入高盐食品中，可以通过改变食品的颜色、表面粘稠度、风味等影响食品各方面的品质。比如在酱油的生产过程中，嗜盐四联球菌是酱醪发酵中后期的嗜盐乳酸菌，与风味物质氨基甲酸乙酯紧密的联系(廖淡宜等，嗜盐四联球菌的分离及其精氨酸代谢，食品与生物技术学报，2016，35：48-53)；比如中度嗜盐菌也已经被作为食品发酵剂在一些盐发酵的食品中广泛使用(赵百锁等.中度嗜盐菌在生物技术中的应用.微生物学通报，2007，34：359-362)。
目前，现有技术中在对蔬菜高盐浓度下嗜盐菌的分析鉴定及其对挥发性成分的影响方面尚未涉及。分离鉴定出腌制蔬菜中的嗜盐菌，探究嗜盐菌的种类，研究这些嗜盐菌对蔬菜挥发性物质的影响将有利于改善腌制蔬菜的口感风味。
发明内容
针对现有技术存在的问题，本发明提供了一株大洋芽孢杆菌及其应用，目的在于解决现有技术中的一部分问题或至少缓解现有技术中的一部分问题。
本发明是这样实现的，一株大洋芽孢杆菌，其特征在于：所述大洋芽孢杆菌保藏编号为CCTCC NO：M 20191070。
如上述的一株大洋芽孢杆菌在腌制蔬菜中的应用。
进一步地，所述应用方法为，将所述大洋芽孢杆菌的菌液接种至腌制蔬菜的卤水中。
进一步地，卤水中所述大洋芽孢杆菌菌液的接种量为6％。
进一步地，所述腌制蔬菜包括襄阳大头菜。
进一步地，将所述大洋芽孢杆菌的菌液接种至襄阳大头菜的卤水中后，37℃恒温培养。
本发明还披露了所述的一株大洋芽孢杆菌在增加腌制蔬菜中的挥发性物质种类和/或含量中的应用。
本发明还披露了所述的大洋芽孢杆菌在减少腌制蔬菜中刺激性辛辣风味物质含量中的应用。
进一步地，所述刺激性辛辣风味物质包括异硫氰酸烯丙酯。
综上所述，本发明的优点及积极效果为：
本发明从襄阳大头菜卤水中筛选出了一株嗜盐菌菌株，经形态、理化和16S rRNA序列鉴定为大洋芽孢杆菌，该菌株能够显著提高腌制蔬菜中挥发性物质种类及含量，降低辛辣味物质的含量，能够改善腌制大头菜的口感风味，具有良好的开发应用前景。
附图说明
图1是空白对照组的卤水中挥发性风味物质的气相色谱图；
图2是添加大洋芽孢杆菌G8(Oceanobacillus picturae G8)作为发酵剂后卤水中挥发性风味物质的气相色谱图。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白，以下结合实施例对本发明进行进一步详细说明，各实施例及试验例中所用的设备和试剂如无特殊说明，均可从商业途径得到。此处所描述的具体实施例仅用以解释本发明，并不用于限定本发明。
本发明披露了一株大洋芽孢杆菌及其应用，本发明涉及的大洋芽孢杆菌名称为墓画大洋芽孢杆菌G8，Oceanobacillus picturae G8，菌株已于2019年12月20日保藏于中国典型培养物保藏中心，地址为中国.武汉.武汉大学，保藏编号为CCTCC NO：M 20191070。该大洋杆菌的获取及应用具体如下各实施例所示。
实施例1菌株的分离与鉴定
1、菌株的分离筛选
以襄阳大头菜卤水为原料，选择高盐LB培养基，吸取1mL卤水样品涂于高盐LB培养基上，37℃恒温培养，再挑取菌落进行平板划线得到单菌落，共得到10种单菌落，接种到襄阳大头菜卤水后测定挥发性成分，筛选出两种能显著增加腌制襄阳大头菜挥发性物质的种类、改善主要挥发性物质的含量的菌株——大洋芽孢杆菌G8(Oceanobacillus picturaeG8)，下述简称为G8。
其中每1000mL高盐LB培养基中含胰蛋白胨10g，酵母膏5g，NaCl 10g，琼脂18g。
2、菌株鉴定
1)菌株的形态特征：大洋芽孢杆菌G8(Oceanobacillus picturae G8)为革兰氏阳性菌，芽孢染色为阳性。G8菌体在高盐LB培养基上呈类似圆，透明，中间凸起，边缘不整齐，光滑。
2)菌株的理化性质：大洋芽孢杆菌G8(Oceanobacillus picturae G8)：葡萄糖、果糖、麦芽糖发酵呈阳性，鼠李糖、蔗糖、甘露糖、乳糖、山梨醇、甘露醇、肌醇发酵呈阴性，阿拉伯糖发酵不定，氧化酶、触酶、V.P试验、产H2S反应呈阳性，淀粉酶、硝酸盐产气、亚硝酸盐产气实验呈阴性，最高耐盐度可达20％，可耐pH6.5、pH8.0。
3)菌株16S rRNA测序结果见SEQ ID NO.1所示。
实施例2G8菌株的应用
分别将菌种G8活化培养即接种到高盐LB培养基，200r/min 37℃摇床过夜，再将菌悬液以6％(V/V)的接种量接种到襄阳大头菜的卤水中，37℃恒温培养30d后采用固相微萃取-气相色谱质谱联用技术测定卤水所含的挥发性成分；同时以未接种任何菌悬液的空白卤水为对照。
所用气相色谱-质谱仪：Agilent7890N-5977B，美国安捷伦公司。采用顶空萃取时间为45min。进样方式：手动进样。进样口：GC。质谱仪：启用。色谱条件：柱型为HP-5MS 30m×250nm×0.25um，载气为He，流速为1.2mL/min，进样温度为250℃，起始柱温40℃，保持1min，以5℃/min到130℃，再以8℃/min到200℃，最后以12℃/min速度升温到250℃，保持7min。质谱条件：离子源温度为230℃，四极杆温度为150℃，电离方式为EI，质量范围为55m/z到550m/z。
图1、图2是检测的气相色谱图。腌制襄阳大头菜卤水中挥发性物质的种类及含量如下：
空白对照卤水中含挥发性成分共34种，其中烷烃类化合物1种，烯烃类1种，酚类0种，醇类1种，酸类1种，醛类3种，苯类0种，酯类5种，酮类7种，醚类5种，氰类2种，胺类3种，杂环类5种。
添加G8菌悬液的卤水中含挥发性成分共43种，其中烷烃类化合物1种，烯烃类3种，酚类1种，醇类1种，酸类0种，醛类4种，苯类0种，酯类9种，酮类8种，醚类6种，氰类1种，胺类3种，杂环类6种。
对具有特殊风味的异硫氰酸烯丙酯用SPSS进行含量显著性分析，结果见表1。
表1异硫氰酸烯丙酯含量显著性分析
| 代表物 | 风味特点 | 添加菌株 | 添加前 | 添加后 | P值 |
| 异硫氰酸烯丙酯 | 刺激性辛辣味 | G8 | 4.19±0.24 | 0.87±0.01 | 0.0330 |
*P>0.05为“不显著”；P≤0.05为“显著”。
异硫氰酸烯丙酯是蔬菜中刺激性辛辣风味的主要来源，在一定外界条件下由芥子酶催化硫代葡萄糖苷形成，使得蔬菜不易于食用，添加G8菌株后显著减小这一成分的含量，使大头菜食用口感更好。因此，本发明筛选的G8菌种可以作为发酵剂接种到蔬菜卤水中，用于丰富、改善腌制蔬菜的口感风味。
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。
序列表
<110> 华中农业大学
<120> 一株大洋芽孢杆菌及其应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1392
<212> DNA
<213> 墓画大洋芽孢杆菌G8 (Oceanobacillus picturae G8)
<400> 1
cagaacggaa ctcttcggag ggaagttcgt ggaacgagcg gcggacgggt gagtaacacg 60
taggcaacct gcctgtaaga ctgggataac tcgcggaaac gcgagctaat accggataac 120
actttccatc tcctgatggg aagttgaaag gcggcttttg ctgtcactta cagatgggcc 180
tgcggcgcat tagctagttg gtggggtaac ggctcaccaa ggcgacgatg cgtagccgac 240
ctgagagggt gatcggccac actgggactg agacacggcc cagactccta cgggaggcag 300
cagtagggaa tcttccgcaa tggacgaaag tctgacggag caacgccgcg tgagtgatga 360
aggttttcgg atcgtaaaac tctgttgtcg gggaagaaca agtacgatag taactgatcg 420
taccttgacg gtacccgacc agaaagccac ggctaactac gtgccagcag ccgcggtaat 480
acgtaggtgg caagcgttgt ccggaattat tgggcgtaaa gcgctcgcag gcggttcttt 540
aagtctgatg tgaaatcttg cggctcaacc gtaaacgtgc attggaaact ggaggacttg 600
agtgcagaag aggagagtgg aattccacgt gtagcggtga aatgcgtaga gatgtggagg 660
aacaccagtg gcgaaggcga ctctctggtc tgtaactgac gctgaggagc gaaagcgtgg 720
ggagcgaaca ggattagata ccctggtagt ccacgccgta aacgatgagt gctaggtgtt 780
agggggtttc cgccccttag tgctgaagtt aacgcattaa gcactccgcc tggggagtac 840
ggccgcaagg ctgaaactca aaagaattga cggggacccg cacaagcggt ggagcatgtg 900
gtttaattcg aagcaacgcg aagaacctta ccaggtcttg acatcctttg acacctctag 960
agatagagtt ttcccttcgg ggacaaagtg acaggtggtg catggttgtc gtcagctcgt 1020
gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc cttgatctta gttgccagca 1080
tttagttggg cactataagg tgactgccgg tgacaaaccg gaggaaggtg gggatgacgt 1140
caaatcatca tgccccttat gacctgggct acacacgtgc tacaatggac ggaacaaagg 1200
gaagcgaacc cgcgaggtcc agcaaatccc ataaaaccgt tctcagttcg gattgcaggc 1260
tgcaactcgc ctgcatgaag ccggaatcgc tagtaatcgc ggatcagcat gccgcggtga 1320
atacgttccc gggtcttgta cacaccgccc gtcacaccac gagagttcgt aacacccgaa 1380
gtcgtagagg ta 1392
Claims (9)
1.一株大洋芽孢杆菌，其特征在于：所述大洋芽孢杆菌名称为墓画大洋芽孢杆菌（Oceanobacillus picturae）G8，保藏编号为CCTCC NO：M 20191070。
2.如权利要求1所述的一株大洋芽孢杆菌在腌制蔬菜中的应用。
3.根据权利要求2所述的一株大洋芽孢杆菌在腌制蔬菜中的应用，其特征在于：所述应用方法为，将所述大洋芽孢杆菌的菌液接种至腌制蔬菜的卤水中。
4.根据权利要求3所述的一株大洋芽孢杆菌在腌制蔬菜中的应用，其特征在于：卤水中所述大洋芽孢杆菌菌液的接种量为6%。
5.根据权利要求3所述的一株大洋芽孢杆菌在腌制蔬菜中的应用，其特征在于：所述腌制蔬菜为襄阳大头菜。
6.根据权利要求5所述的一株大洋芽孢杆菌在腌制蔬菜中的应用，其特征在于：将所述大洋芽孢杆菌的菌液接种至襄阳大头菜的卤水中后，37℃恒温培养。
7.如权利要求1所述的一株大洋芽孢杆菌在增加腌制蔬菜卤水中的挥发性物质种类中的应用。
8.如权利要求1所述的一株大洋芽孢杆菌在减少腌制蔬菜卤水中刺激性辛辣风味物质含量中的应用。
9.根据权利要求8所述的应用，其特征在于，所述刺激性辛辣风味物质包括异硫氰酸烯丙酯。
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| KR101840851B1 (ko) * | 2016-12-21 | 2018-03-21 | 창원대학교 산학협력단 | 신규한 오세아노바실러스 소재 ｂｃｎｕ ３０２５ 균주 |
| CN109825456A (zh) * | 2019-03-15 | 2019-05-31 | 青海省农林科学院 | 一种玛纳斯海洋芽孢杆菌E40208a1及其应用 |
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| KR101840851B1 (ko) * | 2016-12-21 | 2018-03-21 | 창원대학교 산학협력단 | 신규한 오세아노바실러스 소재 ｂｃｎｕ ３０２５ 균주 |
| CN109825456A (zh) * | 2019-03-15 | 2019-05-31 | 青海省农林科学院 | 一种玛纳斯海洋芽孢杆菌E40208a1及其应用 |
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