一株高产细菌素乳酸菌的分离与鉴定

doi:10.11924/j.issn.1000-6850.casb16060059

中国农学通报 ›› 2017, Vol. 33 ›› Issue (10): 150-157.doi: 10.11924/j.issn.1000-6850.casb16060059

一株高产细菌素乳酸菌的分离与鉴定

赵丹,王瑶,杜仁鹏,杨睿睿,王琪,葛菁萍

黑龙江大学生命科学学院/微生物省高校重点实验室;农业微生物技术教育工程研究中心,黑龙江大学生命科学学院/微生物省高校重点实验室;农业微生物技术教育工程研究中心,黑龙江大学生命科学学院/微生物省高校重点实验室;农业微生物技术教育工程研究中心,黑龙江大学生命科学学院/微生物省高校重点实验室;农业微生物技术教育工程研究中心,黑龙江大学生命科学学院/微生物省高校重点实验室;农业微生物技术教育工程研究中心,黑龙江大学生命科学学院/微生物省高校重点实验室;农业微生物技术教育工程研究中心

收稿日期:2016-06-12 修回日期:2017-03-22 接受日期:2016-07-25 出版日期:2017-04-11 发布日期:2017-04-11
通讯作者: 葛菁萍
基金资助:
国家自然科学基金青年科学基金项目“酸菜发酵生态系统中细菌群落与代谢物组特征耦合机制”(31300355)；哈尔滨市科技局青年后备人才项目“功能性乳酸菌发酵酸菜生态学过程研究”(2014RFQXJ101)；黑龙江大学研究生创新科研项目资金“酸菜发酵生态系统中细菌群落与代谢物组特征耦合机制”(YJSCX2015-090HLJU)；黑龙江省政府博士后项目“疣孢漆斑菌(Myrothecium verrucaria)NF-05 漆酶的产生、特性及应用研究” (LBH-Z15214)；黑龙江大学杰出青年基金项目“果胶质降解菌在温水沤麻生态系统中的过程与功能研究”(JCL201305)。

Isolation and Identification of a High Bacteriocin-producing Lactic Acid Bacterium

葛菁萍

Received:2016-06-12 Revised:2017-03-22 Accepted:2016-07-25 Online:2017-04-11 Published:2017-04-11

摘要/Abstract

摘要： 为了满足细菌素的市场需求，降低生产成本，筛选高产细菌素菌株并应用于食品生产成为当前研究的重点。本研究从东北自然酸菜发酵液中分离筛选出一株高产细菌素的乳酸菌11MZ-5-2，发酵48 h 时，细菌素含量达到(344.47±5.50) IU/mL。采用生理生化及分子生物学方法鉴定该菌株，并构建系统发育树。结果表明：菌株11MZ-5-2 为革兰氏阳性细菌，杆状，无芽胞，最适生长温度为30~37℃，耐盐范围为0%~10%，最适pH 5.5~6.5，主要脂肪酸组分为C16:0、C18:0 和C18:1。16S rDNA部分序列长度为1446 bp，与植物乳杆菌(Lactobacillus plantarum) IDCC 3501 相似性最大，达99%，将该菌株命名为L.plantarum 11MZ-5-2。菌株L. plantarum 11MZ-5-2 可作为发酵剂或将细菌素产物应用于食品中，可以有效地防止杂菌污染，延长食品保质期。

关键词: 棚栽嫁接西瓜, 棚栽嫁接西瓜, 黄瓜绿斑驳花叶病毒病, 发生动态, 田间扩展

Abstract: order to meet the market demand of bacteriocin and reduce the production cost, screening a high bacteriocin-producing bacterium and applying it to food production has become the focus of current research. In this study, a lactic acid bacterium 11MZ-5-2 with high bacteriocin-producing ability was isolated from liquid sample of pickle Chinese cabbage. The bacteriocin titer of strain 11MZ-5-2 was (344.47±5.50) IU/mL after 48 h fermentation. Strain 11MZ-5-2 was identified according to physiological and biochemical methods. Meanwhile, the phylogenetic tree was constructed based on its partial 16S rDNA sequence. The results showed that strain 11MZ-5-2 was a kind of rod gram-positive bacterium with no spores, the optimum temperature range was 30-37℃ and the salt-tolerance range was 0%-10%, and the optimum pH range was 5.5-6.5. The main aliphatic acid components of strain 11MZ-5-2 were C16:0, C18:0 and C18:1. The partial sequence of the 16S rDNA of strain 11MZ-5-2 was 1446 bp which was 99% similar withLactobacillus plantarum IDCC 3501. Strain 11MZ-5-2 was identified asL. plantarum .L. plantarum 11MZ-5-2 could be applied as a starter culture in food fermentation. Furthermore, the bacteriocins produced byL. plantarum 11MZ-5-2 could be used as preservatives in food to prevent bacterial pollution and prolong the shelf-life.

中图分类号:

赵丹,王瑶,杜仁鹏,杨睿睿,王琪,葛菁萍. 一株高产细菌素乳酸菌的分离与鉴定[J]. 中国农学通报, 2017, 33(10): 150-157.

葛菁萍. Isolation and Identification of a High Bacteriocin-producing Lactic Acid Bacterium[J]. Chinese Agricultural Science Bulletin, 2017, 33(10): 150-157.

参考文献

[1] 房春红, 刘杰, 许修宏. 乳酸菌素的研究现状和发展趋势[J]. 中国乳品工业, 2006, 34(2): 53-55.
[2] Lizziane K W, Fabrício Luiz T, Elaine C P. Identification of the bacteriocin produced by cheese isolate Lactobacillus paraplantarum FT259 and its potential influence on?Listeria monocytogenes biofilm formation[J]. Food Science and Technology, 2015, 64(2): 586-592.
[3] Weerapong W, Laphaslada P, Amonlaya T, et al. Two putatively novel bacteriocins active against Gram-negative food borne pathogens produced by Weissella hellenica BCC 7293[J]. Food Control, 2015, 55: 176-184.
[4] Lei L and Pinglan L. Complete genome sequence of Lactobacillus paraplantarum L-ZS9, a probiotic starter producing class II bacteriocins[J]. Journal of biotechnology, 2016, 222: 6-15.
[5] 李铁军, 李爱云, 张晓峰. 乳酸菌抗菌机理研究进展[J]. 微生物学报, 2002, 29(5): 81-85.
[6] Sabrina D S S, Michele V, José Manuel D G, et al. Overview of Lactobacillus plantarum as a promising bacteriocin producer among lactic acid bacteria[J]. Food Research International, 2014, 64: 527-536.
[7] 范丽平, 王丽娟, 张海松. 植物乳杆菌的分离鉴定及生长特性研究[J]. 安徽农业科学, 2012, 40(35): 17311-17313.
[8] 段超, 万翠香. 泡菜中植物乳杆菌的筛选及益生活性研究[J]. 食品工业科学, 2015, 36(24): 201-221.
[9] Marta H, Garriga?M, Aymerich T, et al. Biochemical characterization of lactobacilli from dry fermented sausages[J]. International Journal of Food Microbiology, 1993, 18(2): 107-113.
[10] Maria S, Katri H, Tiina M, et al. The effect of lactose derivatives lactulose, lactitol and lactobionic acid on the functional and technological properties of potentially probiotic Lactobacillus strains[J]. International Dairy Journal, 2003, 13(4): 291-302.
[11] Saarela M, Hallama K, Mattil-Sandholm T, et al. The effect of lactose derivatives lactulose, lactitol and lactobionic acid on the functional and technological properties of potentially probiotic Lactobacillus strains[J]. International Dairy Journal, 2003, 13(4): 291-302.
[12] Su Y N, Seok-Seong K, Cheol-Heui Y, et al. Lipoteichoic acid from Lactobacillus plantarum inhibits Pam2CSK4-induced IL-8 production in human intestinal epithelial cells[J]. Molecular Immunology, 2015, 64(1): 183-189.
[13] 武俊瑞, 李欣, 张苗, 等. 自然发酵酸菜汁中乳杆菌的分离鉴定[J]. 食品科学, 2012, 33(15): 191-194.
[14] 布坎南 R E, E吉本斯 N. 伯杰细菌鉴定手册[M]. 9版. 北京: 科学出版社, 1984: 677-821.
[15] 郑雯, 孙琳, 宋诙. 一株植物乳杆菌所产细菌素的性质及诱变筛选[J]. 食品科技, 2014, 39(10): 12-16.
[16] 刘书亮, 张艾青, 田刚, 等. 植物乳杆菌P158的生长曲线及其细菌素的特性[J]. 核农学报, 2009, 23(6): 1021-1025.
[17] 杨晓晖, 籍保平, 李博, 等. 泡菜中优良乳酸菌的分离鉴定及其发酵性能的研究[J]. 食品科学, 2005, 25(5): 130-134.
[18] 东秀珠, 蔡妙英. 常见细菌系统鉴定手册[M]. 北京: 科学出版社, 2001.
[19] 张书光, 张云娟, 代卫东, 等. Viili乳制品中干酪乳杆菌的分离鉴定[J]. 微生物学杂志, 2012, 32(3): 47-52.
[20] 赵玉娟, 牛春华, 张雪, 等. 16S rRNA序列分析及其在乳酸菌分类、鉴定中的应用[J]. 食品科学, 2009, 30(7): 299-303.
[21] 凌代文,东秀珠. 乳酸细菌分类鉴定及实验方法[M]. 北京: 中国轻工业出版社, 1999: 6-38.
[22] 马凯, 李春玲, 程池. Lactobacillus的系统发育分析及分群[J]. 食品与发酵工业, 2007, 33(2): 27-35.
[23] Deeplina D, Arun G. Antioxidant activity and γ-aminobutyric acid (GABA) producing ability of probiotic Lactobacillus plantarum DM5 isolated from Marcha of Sikkim[J]. Food Science and Technology, 2015, 61(1): 263-268.
[24] Hanen B Ayed, Hana M, Noomen H, et al. Isolation and biochemical characterisation of a bacteriocin-like substance produced by Bacillus amyloliquefaciens An6[J]. Journal of Global Antimicrobial Resistance, 2015, 3(4): 255-261.
[25] 张艾青, 刘书亮, 敖灵. 产广谱细菌素乳酸菌的筛选和鉴定[J]. 微生物学通报, 2007, 34(4): 753-756.
[26] Blanca E G, Issac K O C, Scott E M, et al. Effect of Lactococcus lactis UQ2 and its bacteriocin on Listeria monocytogenes biofilms[J]. Food Control, 2008, 19(7): 670-680.
[27] Imran K, Sun Chul K. Probiotic potential of nutritionally improved Lactobacillus plantarum DGK-17 isolated from Kimchi – A traditional Korean fermented food[J]. Food Control, 2016, 60: 88-94.
[28] Fidel T. The role of muscle enzymes in dry-cured meat products with different drying conditions[J]. Trends in Food Science & Technology, 2006, 17(4): 164-168.
[29] 赵辉, 霍贵成, 王葳. 利用植物乳杆菌进行啤酒生物酸化的初步研究[J]. 酿酒科技, 2006, 9: 44-48.
[30] Takashi K, Yurie M, Miho K, et al. Bile acid-lowering properties of Lactobacillus plantarum Sanriku–SU3 isolated from Japanese surfperch fish[J]. Food Bioscience, 2016, 14: 41-46.
[31] Matheus de S B, Svetoslav D Todorov, Iskra I, et al. Improving safety of salami by application of bacteriocins produced by an autochthonous Lactobacillus curvatus isolate[J]. Food Microbiology, 2015, 46: 254-62.
[32] 范远景, 孙锋, 黄云霞, 等. 一株LABR-X植物乳酸杆菌的产条件优化与所产小肽抗菌效果的研究[J]. 食品科学, 2008, 29(11): 383-386.
[33] Todorov S D, Dicks L M T. Effect of medium components on bacteriocin production by Lactobacillus plantarum strains ST23LD and ST341LD, isolated from spoiled olive brine[J]. Microbiological Research, 2006, 161(2): 102-108.
[34] 张杨, 孟祥晨. 自然发酵酸菜中乳杆菌的分离鉴定与多态性分析[J]. 2009, 37(2): 19-22.
[35] Matheus S B, Svetoslav D T, Cynthia H J, et al. Bacteriocin production by Lactobacillus curvatus MBSa2 entrapped in calcium alginate during ripening of salami for control of Listeria monocytogenes[J]. Food Control, 2015, 47: 147-153.
[36] Bouksaim M, Lacroix C, Audet P M, et al. Effects of mixed starter composition on nisin Z production by Lactococcus lactis subsp. lactis biovar. diacetylactis UL 719 during production and ripening of Gouda cheese[J]. International Journal of Food Microbiology, 2000, 59(3): 141-156.
[37] 张子健, 刘玉恩, 赵谦, 等. 植物乳杆菌C8-1产类细菌素的初步研究[J]. 中国乳品工业, 2010, 38(1): 15-22.
[38] Bujalace C, Jimenez-ValerI M, Moreno E, et al. Lack of correlation between in vitro antibiosis and in vivo protection against enteropathogenic bacteria by probiotic lactobacilli[J]. Research Microbiology, 2014, 165(1): 14-20.

一株高产细菌素乳酸菌的分离与鉴定

Isolation and Identification of a High Bacteriocin-producing Lactic Acid Bacterium

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 11

编辑推荐

Metrics

本文评价

关于本刊

作者中心

审者中心

在线期刊

联系我们

[1]	马俊杰, 高有华, 阿布力孜, 郭庆元. 库尔勒地区梨木虱发生动态及黄板诱杀效果研究[J]. 中国农学通报, 2022, 38(24): 109-112.
[2]	赵全新, 高有华, 荣华, 赵志强, 张卢慧, 郭庆元. 库尔勒地区香梨茎蜂发生动态及黄板控害效果[J]. 中国农学通报, 2021, 37(19): 124-127.
[3]	胡翔,陈世春,李品武,林强,鲁礼勋,王晓庆,彭萍. 茶园中柑橘始叶螨及其天敌的发生特点[J]. 中国农学通报, 2018, 34(13): 155-158.
[4]	祝国栋,李朝霞,薛明,赵海朋,刘芳,纪桂霞. 不同设施栽培韭菜韭蛆的发生规律及其影响的环境因素[J]. 中国农学通报, 2017, 33(10): 134-139.
[5]	蔡美艳,周琳娜,李程巧,冯永斌,叶建人. 棚栽嫁接西瓜种栽地土壤黄瓜绿斑驳花叶病毒病带毒性研究[J]. 中国农学通报, 2016, 32(7): 97-103.
[6]	叶建人,蔡美艳,冯永斌,陈海波,周琳娜. 棚栽嫁接西瓜黄瓜绿斑驳花叶病毒病发病动态及其田间扩展研究初报[J]. 中国农学通报, 2015, 31(16): 166-172.
[7]	杨晓贺. 2013年三江平原地区大豆蚜及其天敌种群发生动态[J]. 中国农学通报, 2014, 30(16): 278-281.
[8]	韩凤英杨慧秦旭王绍敏杨向黎?. 济南市紫花苜蓿害虫和天敌种类及其发生动态的研究[J]. 中国农学通报, 2012, 28(29): 5-9.
[9]	余金咏1,周印富2,于泉林1,齐慧霞1,贺子典1. 亚洲玉米螟发生动态及释放松毛虫赤眼蜂防治效果[J]. 中国农学通报, 2009, 25(24): 344-351.
[10]	刘水芳,于福安,顾红艳,魏立军,张建民,孙淑琴,杨秀荣. 天津稻区水稻条纹叶枯病发生动态与综合防治[J]. 中国农学通报, 2007, 23(5): 302-302.
[11]	汪恩国,王华弟,关梅萍,陈克松. 杂交水稻黑条矮缩病发生为害损失与防治指标研究[J]. 中国农学通报, 2005, 21(1): 278-278.