食品与发酵工业 >

2022 , Vol. 48 >Issue 17: 87 - 92

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.032080

研究报告

溶源性发酵乳杆菌噬菌体的诱导及灭活方法

  • 郭蛇 ,
  • 许铭 ,
  • 张灿 ,
  • 朱含芳 ,
  • 陈霞
展开
  • (内蒙古农业大学, 乳品生物技术与工程教育部重点实验室,农业农村部奶制品加工重点实验室,内蒙古自治区乳品生物技术与工程重点实验室,内蒙古 呼和浩特,010018)
第一作者:硕士研究生(陈霞教授为通信作者,E-mail:chenxia8280@163.com)

收稿日期: 2022-04-21

  修回日期: 2022-05-12

  网络出版日期: 2022-10-01

基金资助

国家自然科学基金项目(32160550,31760447,31301517);中央引导地方项目(2021ZY0022);内蒙古自然科学基金项目(2021MS03014,2019BS03003)

收起

Induction and inactivation method of lysogenic phage of Lactobacillus fermentum

  • GUO She ,
  • XU Ming ,
  • ZHANG Can ,
  • ZHU Hanfang ,
  • CHEN Xia
Expand
  • (Key Laboratory of Dairy Biotechnology and Engineering of Ministry of Education, Key Laboratory of Dairy Processing of Ministry of Agriculture and Rural Areas, Key Laboratory of Dairy Biotechnology and Engineering of Inner Mongolia Autonomous Region, Inner Mongolia Agricultural University, Hohhot 010018,China)

Received date: 2022-04-21

  Revised date: 2022-05-12

  Online published: 2022-10-01

Fold

摘要

乳杆菌溶源性噬菌体在乳品发酵工业中分布广泛,其侵染特性特殊,具有一定的隐蔽性,但在一定条件下,可能被诱导转变为烈性噬菌体,使发酵剂菌株裂解,导致发酵失败。因此,溶源性噬菌体已成为发酵生产过程中的主要安全隐患之一。该研究以发酵乳杆菌为研究对象,利用不同诱导方式对其诱导,并评价热处理及不同化学杀菌剂对诱导出噬菌体的灭活效果。结果显示,发酵乳杆菌(Lactobacillus fermentum)IMAU32579经1.9 μg/mL的丝裂霉素C诱导后,成功获得噬菌体,将其命名为LFP-VB1。形态学分析显示,该噬菌体属于长尾噬菌体科。该噬菌体对热敏感,63 ℃处理4 min即完全失活。化学杀菌剂对其灭活效果有限,800 mg/L NaClO溶液处理60 min仅使其下降6.29个对数级。

关键词： 发酵乳杆菌; 溶源性噬菌体; 诱导; 灭活; 化学消毒剂; 热处理

本文引用格式

郭蛇 , 许铭 , 张灿 , 朱含芳 , 陈霞 . 溶源性发酵乳杆菌噬菌体的诱导及灭活方法[J]. 食品与发酵工业, 2022 , 48(17) : 87 -92 . DOI: 10.13995/j.cnki.11-1802/ts.032080

Abstract

Lysogenic phages of Lactobacillus spp. are widely distributed in dairy fermentation setting. Their special infection characteristics are somewhat hidden and arduous to detect. However, under certain conditions, these phages could cause lysis of fermenting lactic acid bacteria strains, and eventually lead to fermentation failure. Therefore, lysogenic phages have become one of the major safety hazards in dairy fermentation. The Lactobacillus fermentum IMAU32579 was induced and the inactivation effects of heat treatment and different chemical biocides were evaluated. The results showed that a bacteriophage named as LFP-VB1was successful induced from L. fermentum IMAU32579 after treatment with 1.9 μg/mL mitomycin C. Morphology analysis revealed that this bacteriophage strain belonged to the family of Siphoviridae. It was observed that bacteriophage LFP-VB1 was sensitive to heat treatment (completely inactivated 63℃ for 4 min). The inactivation effects of biocides were insignificant, 6.29 lg decrease was observed after treatment with 800 mg/L NaClO for 60 min. This study can provide some theoretical basis for the constructing of phage control measures.

Key words： Lactobacillus fermentum; lysogenic phage; induction; inactivation; chemical biocide; heat treatment

参考文献

[1] SOLOVEVA I V, NOVIKOVA N A, TOCHILINA A G, et al.The probiotic strain Lactobacillus fermentum 39:Biochemical properties, genomic features, and antiviral activity[J].Microbiology, 2021, 90(2):219-225.
[2] 张秀红. 溶源性发酵乳杆菌及其温和噬菌体的研究[D].济南:山东大学, 2006.
ZHANG X H.Studies on lysogenic Lactobacillus fermentum and its temperate bacteriophages[D].Jinan:Shandong University, 2006.
[3] 赵岩. 中国人群肠道中发酵乳杆菌的基因差异及其对肠道的生理调节作用[D].无锡:江南大学, 2021.
ZHAO Y.Genetic difference and intestinal regulatory function of Limosilactobacillus fermentum in Chinese population[D].Wuxi:Jiangnan University, 2021.
[4] ZHAO Y, YU L L, TIAN F W, et al.An optimized culture medium to isolate Lactobacillus fermentum strains from the human intestinal tract[J].Food & Function, 2021, 12(15):6 740-6 754.
[5] GARNEAU J E, MOINEAU S.Bacteriophages of lactic acid bacteria and their impact on milk fermentations[J].Microbial Cell Factories, 2011, 10(Suppl 1):S20.
[6] 周园, 王岚, 李旗, 等.噬菌体在鲍曼不动杆菌耐药过程中的作用[J].沈阳医学院学报, 2013, 15(3):140-141, 144.
ZHOU Y, WANG L, LI Q, et al.The effect of bacteriophages on drug-resistance of Acinetobacter baumannii[J].Journal of Shenyang Medical College, 2013, 15(3):140-141;144.
[7] SUÁREZ V B, REINHEIMER J A.Effectiveness of thermal treatments and biocides in the inactivation of Argentinian Lactococcus lactis phages[J].Journal of Food Protection, 2002, 65(11):1 756-1 759.
[8] MASKE B L, DE MELO PEREIRA G V, DA SILVA VALE A, et al.Viruses in fermented foods:Are they good or bad? Two sides of the same coin[J].Food Microbiology, 2021, 98:103794.
[9] MAHONY J, MOSCARELLI A, KELLEHER P, et al.Phage biodiversity in artisanal cheese wheys reflects the complexity of the fermentation process[J].Viruses, 2017, 9(3):45.
[10] 柴诗语, 郭静, 马瑞瑞, 等.食品工业中噬菌体灭活方法研究进展[J].中国乳品工业, 2019, 47(11):36-40.
CHAI S Y, GUO J, MA R R, et al.Inactivation methods of bacteriophages in food industry[J].China Dairy Industry, 2019, 47(11):36-40.
[11] FAN C, FANG C, ZHANG Y, et al.A new method for determining the frequency of induction of λ prophage[J].Virologica Sinica, 2002, 17(4):367-370.
[12] 郭静. 溶原性芽孢杆菌噬菌体的生物学特性及其灭活方法[D].呼和浩特:内蒙古农业大学, 2019.
GUO J.Biological characteristics and inactivation methods of lysogenic Bacillus phage[D].Hohhot:Inner Mongolia Agricultural University, 2019.
[13] 杨敏. 水稻白叶枯病菌前噬菌体高效释放的诱导剂鉴定及转座酶在其中的功能研究[D].杭州:浙江大学, 2020.
YANG M.Identification of an inducer for high-efficiency prophage release of Xanthomonas oryzae pv.oryzae and functional analysis of associated transposases[D].Hangzhou:Zhejiang University, 2020.
[14] HO C H, STANTON-COOK M, BEATSON S A, et al.Stability of active prophages in industrial Lactococcus lactis strains in the presence of heat, acid, osmotic, oxidative and antibiotic stressors[J].International Journal of Food Microbiology, 2016, 220:26-32.
[15] 季晖. 药理学[M].2版.南京:东南大学出版社, 2019.
[16] PEI Z M, SADIQ F A, HAN X, et al.Identification, characterization, and phylogenetic analysis of eight new inducible prophages in Lactobacillus[J].Virus Research, 2020, 286:198003.
[17] MAHONY J, VAN SINDEREN D.Current taxonomy of phages infecting lactic acid bacteria[J].Frontiers in Microbiology, 2014, 5:7.
[18] TRUCCO V, REINHEIMER J, QUIBERONI A, et al.Adsorption of temperate phages of Lactobacillus delbrueckii strains and phage resistance linked to their cell diversity[J].Journal of Applied Microbiology, 2011, 110(4):935-942.
[19] 林泽永, 梁倩雯, 苏燧, 等.一株短乳杆菌烈性噬菌体的分离鉴定与生化特性[J].食品与发酵工业, 2022,48(12):226-232.
LIN Z Y, LIANG Q W, SU S, et al.Isolation, identification and biochemical characterization of a virulent phage strain of Lactobacillus sp[J].Food and Fermentation Industries, 2022,48(12):226-232.
[20] 吴淑梅, 吴清, 王晓波, 等.乙醇复方手消毒液的消毒性能及毒性观察[J].中国消毒学杂志, 2009, 26(6):637-639.
WU S M, WU Q, WANG X B, et al.Observation on toxicity and disinfection function of glycerol-alcohol hand-washing solution[J].Chinese Journal of Disinfection, 2009, 26(6):637-639.
[21] 段晓杰, 王萌, 陈丽媛, 等.乙醇工艺对同种异体肌腱病毒灭活效果的研究[J].中国药事, 2021, 35(5):544-551.
DUAN X J, WANG M, CHEN L Y, et al.Study on the effect of ethanol process on virus inactivation of allogeneic tendon[J].Chinese Pharmaceutical Affairs, 2021, 35(5):544-551.
[22] SECHAUD L, CLUZEL P J, ROUSSEAU M, et al.Bacteriophages of lactobacilli[J].Biochimie, 1988, 70(3):401-410.
[23] CAPRA M L, QUIBERONI A, REINHEIMER J A.Thermal and chemical resistance of Lactobacillus casei and Lactobacillus paracasei bacteriophages[J].Letters in Applied Microbiology, 2004, 38(6):499-504.
[24] QUIBERONI A, SUÁREZ V B, REINHEIMER J A.Inactivation of Lactobacillus helveticus bacteriophages by thermal and chemical treatments[J].Journal of Food Protection, 1999, 62(8):894-898.
[25] BINETTI A G, REINHEIMER J A.Thermal and chemical inactivation of indigenous Streptococcus thermophilus bacteriophages isolated from Argentinian dairy plants[J].Journal of Food Protection, 2000, 63(4):509-515.
[26] BRIGGILER MARCÓ M, DE ANTONI G L, REINHEIMER J A, et al.Thermal, chemical, and photocatalytic inactivation of Lactobacillus plantarum bacteriophages[J].Journal of Food Protection, 2009, 72(5):1 012-1 019.
Options
文章导航

/

〈 〉

技术支持：北京玛格泰克科技发展有限公司