CN112280715A - 一株新型食醋污染菌及其分离方法 - Google Patents
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Abstract
本发明涉及一种野生型菌株及其分离方法,具体涉及一株新型食醋污染菌及其分离方法,特别涉及导致食醋胀气变质的污染菌。一株新型食醋污染菌为乳杆菌Lactobacillus sp,菌株编号为Z‑1,保藏于中国普通微生物菌种保藏管理中心,保藏号为CGMCC No.20399,保藏日期为2020年7月20日。本发明提供的Lactobacillus sp.Z‑1筛选自胀气变质食醋,菌落呈圆形,米黄色,表面光滑湿润,边缘规则,中央微凸起。形态和分子生物学鉴定为一株野生型食醋污染菌乳杆菌Lactobacillus sp.Z‑1,乳杆菌Lactobacillus sp.Z‑1是一株新种,耐酸能力强,在pH值3.5环境条件下生长良好,能导致食醋发生胀壶变质现象。
Description
技术领域
本发明涉及一种微生物及其分离方法,具体涉及一株新型食醋污染菌及其分离方法,特别涉及导致食醋胀气变质的污染菌。
背景技术
食醋作为我国传统酸性调味品,拥有三千多年的发展历史,其生产离不开微生物的催化,随着食醋发酵产业的不断发展,从自然菌群到单一菌株再到多菌种的使用,微生物在食醋发酵的过程中占据着越来越重要的地位。但微生物同时也是影响食醋品质的极大威胁因素,有害微生物的污染和生长,不仅会影响产品质量还会对人们身体健康造成危害。
中国固态发酵食醋的生料发酵和开放式的发酵过程,极易引入环境中微生物,导致成品食醋腐败变质。近年来,食醋行业频发胀壶变质现象,整个产业经济受到一定程度影响。关于食醋中产气微生物的研究不系统,产气微生物种类不明和难常规培养、防控方法缺乏针对性等问题已成为食醋行业亟待解决的技术难题。采用传统培养与高通量测序相结合的方法对食醋产气污染菌进行研究,高通量测序结果表明引起食醋胀气变质的微生物为乳杆菌属,但通过固体培养基分离纯化未能得到菌落,产气菌株受到食醋影响会发生变化,普通培养基可能无法分离,因此微生物分离方法需要改进。研究导致食醋胀气的新型污染微生物对于食醋行业产气微生物的防控有重要意义。
发明内容
本发明目的在于提供一株导致食醋胀气变质的新型污染菌乳杆菌Lactobacillus sp. Z-1。
本发明的第二目的在于提供一株导致食醋胀气变质的新型污染菌乳杆菌Lactobacillus sp. Z-1的分离方法。
所述一株导致食醋胀气变质的新型污染菌为产气耐酸的乳杆菌Lactobacillus sp. Z-1,是一株新种,菌落呈圆形,米黄色,表面光滑湿润,边缘规则,中央微凸起。已于2020年7月20日保藏于中国普通微生物菌种保藏管理中心,地址:北京市朝阳区北辰西路1号院,邮编:100101,保藏中心登记入册编号:CGMCC No. 20399。
所述一株导致食醋胀气变质的新型污染菌为产气耐酸的乳杆菌Lactobacillus sp. Z-1,筛选自四川某胀壶变质食醋中,样品编号D-2,表现出胀壶现象,醋液发粘变黄,醋香弱,伴有淡腐味。
所述一株导致食醋胀气变质的新型污染菌乳杆菌Lactobacillus sp. Z-1的分离方法包括以下步骤:
1)取150 mL锥形瓶,洗净后加入50 mL改良MRS液体培养基,胶塞封口,121℃高温灭菌20min,待体系冷却至室温后,在超净工作台补加复合维生素B溶液,调节pH至3.5。
改良MRS液体培养基配方:葡萄糖20 g/L,蛋白胨10 g/L,牛肉膏10 g/L,酵母浸出粉5 g/L,磷酸氢二钾2 g/L,乙酸钠5 g/L,硫酸镁0.58 g/L,硫酸锰0.25 g/L,吐温-80 2g/L,柠檬酸二铵2 g/L,麸皮浸出液100 mL/L,复合氨基酸1 g/L,核苷酸0.5 g/L;改良MRS固体培养基即添加2%琼脂粉,121℃灭菌15min;改良MRS液体培养基中复合维生素B终浓度为0.5 g/L,过滤除菌。调节pH至3.5在超净工作台用过滤除菌后乳酸:乙酸=1:1调节改良MRS液体培养基pH至3.5。
2)取1 mL变质食醋样品,加入带50 mL改良MRS液体培养基的150 mL锥形瓶中,33℃下静置培养3 d。
3)取样涂布于改良MRS固体培养基平板上,置于厌氧罐中33℃培养一周后,形成肉眼可见的菌落,经反复分离纯化,得到菌株Z-1。
4)将乳杆菌Lactobacillus sp. Z-1接种于塑瓶装的高温灭菌6°-7°食醋中,拧紧瓶盖,培养7-14天,塑瓶膨胀,发生明显产气胀瓶现象。
本发明提取菌株Z-1的基因组DNA并以此为模板采用通用引物27F和1492R扩增16SrDNA,对扩增产物测序,并在NCBI上选取高相似度序列,用MEGA-X软件构建系统发育树。
所述一株导致食醋胀气变质的污染菌乳杆菌Lactobacillus sp.Z-1的16S rDNA序列为:TCGCCCTTGAGAGTTTGATCCTGGCTCAGGATGAACGCTGGCGGTGTGCCTAATACATGCAAGTCGCGCGCACTCCCGTAGATGATTTTGATGCTCTGCATCAAATGATTCTACATTCGAGTGAGCGGCGGATGGGTGAGTAACACGTGGGTAACTTGCCCTTGAGTCTGGGATAGCATCTGGAAACGGATGATAATACCGGATAACAACTGAAACCACATGGTTTCAGTTTGAAAGGCGCGTTTGCGTCGCTCTTGGAGGGACCCGCGGTCCATTAGTTAGTTGGTAAGATAAAGGCCTACCAAGACGATGATGGATAGCCGAACTGAGAGGTTAATCGGCCACATTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTTCACAATGGGCGAAAGCCTGATGAAGCAACACCGCGTGAATGAAGAAGGGTTTCGGCTCGTAAAATTCTGTTGCTAGAGAAGAACGTACCAAGTAGGAAATGGCTTGGTAGTGACGGTATCTGGTTAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTGTTTAAGTCCGATGTGAAAGCCTTCGGCTTAACCGAAGAAGTGCATTGGAAACTGAGCGACTTGAGTACAGAAGAGGACAGTAGAACACCATGTGTAGCGGTGAAATGCGTAGATATATGGTAGAATACCAGTGGCGAAAGCGGCTGTCTGGTCTGTCACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATGCTGTAAACGTTGAGTGCTAGGTGTTAGAGGATTTCCGTCCTTTAGTGCCGAAGTTAACACATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGTGGTGGAGCATGTGGTTTAATTCGATGCTACGCGAAGAACCTTACCAGGCCTTGACATACTTCTGTTAGGCTAAGAGATTAGCTGTTCCCTTCGGGGACAGATCTACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTG
GGTCAAGTCCCGCAACGAGCACAACCCTTGTGACTAGTTGCCAGCATTCAGTTGGGCACTCTAGTCAGACTGCCGGTGATAAACCGGAGGAAGGTGAGGATGACGTCTGATCATCATGCCCCTTATGGCCTGGGCTACACACGTGCTACAATGGACAGTACAATGGGTTGCGAAACCGCGAGGTCAAGCTAATCCCATAAAGCTGTTCTCAGTTCGGACTGCAGTCTGCAACTCGACTGCACGAAGCTGGAATCACTAGTAATCGTGGATCAGCATGCCACGGTGAATACGTTCCCGGGTCTTGTACATACCGCCCGTCACACCATGAGAGTCCGTAACACCCAAAGATAGTGCGGCAACCTCTTTTGAGGAGCCAGCTATCTAAGGTGGGACGAATGATTAGGGTGAAGTCGTAACAAGGTAGCCGTAAAGGGCGACACGCGAATTCGATATCAAGCTTCAGGTCTGCAGTCAATACGA。
附图说明
图1是乳杆菌Lactobacillus sp. Z-1的菌落形态和显微形态图。
图2为乳杆菌Lactobacillus sp. Z-1的系统发育树。
具体实施方式
以下实施例将结合附图对本发明作进一步的说明。
实施例1:菌株的分离
(1)胀壶食醋样品取自四川某麸醋厂,样品编号D-2,表现出胀壶现象,醋液发粘变黄,醋香弱,伴有淡腐味。样品经低温保存后运送至实验室进行下一阶段研究工作。
(2)取150 mL锥形瓶,洗净后加入50 mL改良MRS培养基,胶塞封口,121℃高温灭菌20min,待体系冷却至室温后,在超净工作台补加复合维生素B溶液,调节pH至3.5。取1 mL变质食醋样品,加入带50 mL改良MRS培养基的150 mL锥形瓶中,33℃下静置培养3 d;取样涂布于改良MRS固体培养基平板上,置于厌氧罐中33℃培养一周后,形成肉眼可见的菌落,经反复分离纯化,得到纯菌Z-1。将纯菌Z-1接种于塑瓶装经过高温灭菌的6°-7°食醋中,拧紧瓶盖,培养7-14天,塑瓶膨胀,发生明显产气胀瓶现象。
实施例2:对菌株Z-1进行鉴定
(1)革兰氏染色、过氧化氢酶实验
革兰氏染色:挑取平板上的菌落进行涂片、固定、结晶紫初染、媒染、脱色、番红复染,干燥,镜检;过氧化氢酶实验:待测菌先于空气中暴露30min,用毛细管吸取少量3% H2O2滴于平板表面所生长的菌落。如不产气泡则为过氧化氢酶阴性。
(2)菌落形态、菌体显微形态观察及生理生化鉴定
肉眼观察菌株的菌落形态特征,观察结果见图1a,该菌株呈圆形,米黄色,表面光滑湿润,边缘规则,中央微凸起。革兰氏染色后,于光学显微镜油镜下观察菌株的细胞形态特征,观察结果见图1b,其细胞形态呈杆状,革兰氏染色阳性,过氧化氢酶阴性。
(3)16S rDNA序列分析鉴定
提取该菌的基因组DNA并以此为模板采用通用27F,1492R引物扩增16S rDNA片段,并在NCBI上选取高相似度序列,用MEGA-X计算出序列的系统进化距离,构建系统发育树,参见图2;将16S rDNA测序结果与基因库中序列进行同源性比对,并结合形态学和生理生化指标,最终将菌株Z-1鉴定为乳杆菌Lactobacillus sp.。
以上的实例仅仅是对本发明的实施方式进行描述,而并非对本发明的范围进行限定,对于本领域的技术人员来说,可以对上述说明进行改进或变形,但所有的这些改进或变形均应落入本发明的权利要求确定的保护范围。
序列表
<110> 四川农业大学
<120> 一株新型食醋污染菌及其分离方法
<141> 2020-11-06
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1600
<212> DNA
<213> 乳杆菌(Lactobacillus sp)
<400> 1
tcgcccttga gagtttgatc ctggctcagg atgaacgctg gcggtgtgcc taatacatgc 60
aagtcgcgcg cactcccgta gatgattttg atgctctgca tcaaatgatt ctacattcga 120
gtgagcggcg gatgggtgag taacacgtgg gtaacttgcc cttgagtctg ggatagcatc 180
tggaaacgga tgataatacc ggataacaac tgaaaccaca tggtttcagt ttgaaaggcg 240
cgtttgcgtc gctcttggag ggacccgcgg tccattagtt agttggtaag ataaaggcct 300
accaagacga tgatggatag ccgaactgag aggttaatcg gccacattgg aactgagaca 360
cggtccagac tcctacggga ggcagcagta gggaatcttt cacaatgggc gaaagcctga 420
tgaagcaaca ccgcgtgaat gaagaagggt ttcggctcgt aaaattctgt tgctagagaa 480
gaacgtacca agtaggaaat ggcttggtag tgacggtatc tggttagaaa gtcacggcta 540
actacgtgcc agcagccgcg gtaatacgta ggtggcaagc gttatccgga tttattgggc 600
gtaaagcgag cgcaggcggt tgtttaagtc cgatgtgaaa gccttcggct taaccgaaga 660
agtgcattgg aaactgagcg acttgagtac agaagaggac agtagaacac catgtgtagc 720
ggtgaaatgc gtagatatat ggtagaatac cagtggcgaa agcggctgtc tggtctgtca 780
ctgacgctga ggctcgaaag catgggtagc aaacaggatt agataccctg gtagtccatg 840
ctgtaaacgt tgagtgctag gtgttagagg atttccgtcc tttagtgccg aagttaacac 900
attaagcact ccgcctgggg agtacgaccg caaggttgaa actcaaagga attgacgggg 960
acccgcacaa gtggtggagc atgtggttta attcgatgct acgcgaagaa ccttaccagg 1020
ccttgacata cttctgttag gctaagagat tagctgttcc cttcggggac agatctacag 1080
gtggtgcatg gtcgtcgtca gctcgtgtcg tgagatgttg ggtcaagtcc cgcaacgagc 1140
acaacccttg tgactagttg ccagcattca gttgggcact ctagtcagac tgccggtgat 1200
aaaccggagg aaggtgagga tgacgtctga tcatcatgcc ccttatggcc tgggctacac 1260
acgtgctaca atggacagta caatgggttg cgaaaccgcg aggtcaagct aatcccataa 1320
agctgttctc agttcggact gcagtctgca actcgactgc acgaagctgg aatcactagt 1380
aatcgtggat cagcatgcca cggtgaatac gttcccgggt cttgtacata ccgcccgtca 1440
caccatgaga gtccgtaaca cccaaagata gtgcggcaac ctcttttgag gagccagcta 1500
tctaaggtgg gacgaatgat tagggtgaag tcgtaacaag gtagccgtaa agggcgacac 1560
gcgaattcga tatcaagctt caggtctgca gtcaatacga 1600
Claims (7)
1.一株导致食醋胀壶变质的新型污染菌为乳杆菌Lactobacillus sp. Z-1,其特征在于保藏于中国普通微生物菌种保藏管理中心,保藏地点为北京市朝阳区北辰西路1号院,保藏号为CGMCC No. 20399,保藏日期为2020年7月20日。
2.如权利要求1所述一株乳杆菌Lactobacillus sp. Z-1,其特征为:菌落呈圆形,米黄色,表面光滑湿润,边缘规则,中央微凸起,革兰氏染色阳性,根据菌体形态及系统发育树鉴定为Lactobacillus sp.。
3. 如权利要求1所述一株乳杆菌Lactobacillus sp. Z-1的分离方法,其特征在于包括以下步骤:
1)取150 mL锥形瓶,加入50 mL改良MRS液体培养基,胶塞封口,121℃灭菌20min,待体系冷却至室温后,在超净工作台补加复合维生素B溶液,调节pH至3.5;
2)取1 mL变质食醋样品,加入带50 mL改良MRS液体培养基的150 mL锥形瓶中,33℃下静置培养3 d;
3)取样涂布于改良MRS固体培养基平板上,置于厌氧罐中33℃培养一周后,形成肉眼可见的菌落,经反复分离纯化,得到一株乳杆菌Lactobacillus sp. Z-1;
4)将乳杆菌Lactobacillus sp. Z-1接种于塑瓶装的高温灭菌6°-7°食醋中,拧紧瓶盖,培养7-14天,塑瓶膨胀,发生明显产气胀瓶现象。
4. 如权利要求3所述一株乳杆菌Lactobacillus sp. Z-1的分离方法,其特征在于步骤1)中,所述改良MRS液体培养基配方为:葡萄糖20 g/L,蛋白胨10 g/L,牛肉膏10 g/L,酵母浸出粉5 g/L,磷酸氢二钾2 g/L,乙酸钠5 g/L,硫酸镁0.58 g/L,硫酸锰0.25 g/L,吐温-80 2 g/L,柠檬酸二铵2 g/L,麸皮浸出液100 mL/L,复合氨基酸1 g/L,核苷酸0.5 g/L。
5.所述改良MRS液体培养基中复合维生素B终浓度为0.5 g/L,使用前需要0.22μm滤膜过滤除菌即可。
6. 如权利要求3所述一株乳杆菌Lactobacillus sp. Z-1的分离方法,其特征在于在步骤1)中,所述调节pH至3.5即在超净工作台用0.22μm滤膜过滤除菌后的乳酸:乙酸=1:1调节改良MRS液体培养基pH至3.5。
7. 如权利要求3所述一株乳杆菌Lactobacillus sp. Z-1的分离方法,其特征在于在步骤3)中,所述厌氧固体平板在改良MRS液体培养基的基础上添加2.0%琼脂粉。
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Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1995316A (zh) * | 2006-11-22 | 2007-07-11 | 天津科技大学 | 一种澄清不变质食醋(保健醋)制备工艺 |
CN104774734A (zh) * | 2015-04-09 | 2015-07-15 | 福建师范大学 | 一种富含γ-氨基丁酸的红曲醋制备方法 |
CN106190893A (zh) * | 2016-07-11 | 2016-12-07 | 江苏恒顺醋业股份有限公司 | 一株适用于食醋酿造的发酵乳杆菌及其菌粉的制备方法和应用 |
CN107607585A (zh) * | 2017-10-18 | 2018-01-19 | 山西省食品工业研究所 | 一种利用电子鼻传感器组合优化法监控食醋半成品质量的方法 |
CN109266715A (zh) * | 2018-09-21 | 2019-01-25 | 佛山市海天(江苏)调味食品有限公司 | 一种料酒中产气菌的培养基和检测方法 |
CN109929906A (zh) * | 2019-04-11 | 2019-06-25 | 千禾味业食品股份有限公司 | 一种食醋中产气嗜酸菌的检测方法 |
CN112980980A (zh) * | 2021-04-23 | 2021-06-18 | 江南大学 | 一种用于特异性定量金山醋酸乳杆菌的分子标记和试剂盒以及应用 |
CN113604402A (zh) * | 2021-08-30 | 2021-11-05 | 江苏恒顺醋业股份有限公司 | 一种特异性的乳酸菌培养基及其培养方法和应用 |
CN115236327A (zh) * | 2022-04-25 | 2022-10-25 | 四川农业大学 | 一株新型食醋污染菌的间接elisa检测方法 |
-
2020
- 2020-11-06 CN CN202011227512.3A patent/CN112280715B/zh active Active
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1995316A (zh) * | 2006-11-22 | 2007-07-11 | 天津科技大学 | 一种澄清不变质食醋(保健醋)制备工艺 |
CN104774734A (zh) * | 2015-04-09 | 2015-07-15 | 福建师范大学 | 一种富含γ-氨基丁酸的红曲醋制备方法 |
CN106190893A (zh) * | 2016-07-11 | 2016-12-07 | 江苏恒顺醋业股份有限公司 | 一株适用于食醋酿造的发酵乳杆菌及其菌粉的制备方法和应用 |
CN107607585A (zh) * | 2017-10-18 | 2018-01-19 | 山西省食品工业研究所 | 一种利用电子鼻传感器组合优化法监控食醋半成品质量的方法 |
CN109266715A (zh) * | 2018-09-21 | 2019-01-25 | 佛山市海天(江苏)调味食品有限公司 | 一种料酒中产气菌的培养基和检测方法 |
CN109929906A (zh) * | 2019-04-11 | 2019-06-25 | 千禾味业食品股份有限公司 | 一种食醋中产气嗜酸菌的检测方法 |
CN112980980A (zh) * | 2021-04-23 | 2021-06-18 | 江南大学 | 一种用于特异性定量金山醋酸乳杆菌的分子标记和试剂盒以及应用 |
CN113604402A (zh) * | 2021-08-30 | 2021-11-05 | 江苏恒顺醋业股份有限公司 | 一种特异性的乳酸菌培养基及其培养方法和应用 |
CN115236327A (zh) * | 2022-04-25 | 2022-10-25 | 四川农业大学 | 一株新型食醋污染菌的间接elisa检测方法 |
Non-Patent Citations (3)
Title |
---|
MIN LI等: "Isolation and identification of high-temperature tolerant spoilage microbes during vinegar brewing and growth characterization of the bacterial consortium TYF-LIM-L09 with a srong gas-producing ability.", vol. 39, no. 39, pages 95 - 103 * |
于学健;胡海蓉;曹艳花;程坤;辛亮;张哲;姚粟;: "乳杆菌属分类学地位变迁后菌种名称英解汉译检索表(三)", 食品与发酵工业, no. 17 * |
翟磊等: "一株食醋污染菌CICC 10774 的鉴定及其生长代谢特性", vol. 43, no. 43, pages 1524 * |
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