JIANG Wei, SHEN Wenxiang, ZHENG Juanshan, et al. Mutation Breeding of Lactobacillus rhamnosus from Dairy Cow by 12C6+ Heavy Ion Beam[J]. Science and Technology of Food Industry, 2022, 43(17): 140−148. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021110356.
Citation: JIANG Wei, SHEN Wenxiang, ZHENG Juanshan, et al. Mutation Breeding of Lactobacillus rhamnosus from Dairy Cow by 12C6+ Heavy Ion Beam[J]. Science and Technology of Food Industry, 2022, 43(17): 140−148. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021110356.

Mutation Breeding of Lactobacillus rhamnosus from Dairy Cow by 12C6+ Heavy Ion Beam

More Information
  • Received Date: November 29, 2021
  • Available Online: July 02, 2022
  • In this study, 12C6+ heavy ion beam released from heavy ion accelerator of Chinese Academy of Sciences was used as the radiation mutagenesis source. With acid spot value and bacteriostatic ring value as indexes, functional mutagenesis of Lactobacillus rhamnosus JF12-1 was carried out to determine the optimal radiation dose of mutagenesis through mortality rate and positive and negative mutation rate. The mutated strains under the optimal irradiation dose were screened by acid patch method and screened by inhibition zone method. Then the genetic stability of the genetically stable strains was determined by detecting the change of lactic acid content after continuous passage. 16S rDNA sequencing was performed to locate the mutation sites of the genetically stable strains. The results showed that the mortality rate, positive mutation rate and negative mutation rate were 79.86%, 30.33% and 5.38%, respectively, when the irradiation dose was 300 Gy, which was the best mutagenesis dose. The results showed that the HC value of 20 mutant strains increased by more than 25% compared with the original wild strain. The bacteriostatic activity of 8 strains increased by 15% compared with the original wild strains. Genetic stability analysis showed that the 8 mutant Lactobacillus strains were stable in producing lactic acid. 16S rDNA sequencing revealed that the mutated sites of JF12-1 were not in 16S rRNA gene, and the mutated sites promoting acid production and bacteriostatic performance of JF12-1 were probably in other gene segments. A functional Lactobacillus rhamnosus stable strain with high lactic acid yield and good bacteriostatic activity in vitro was successfully cultured by 12C6+ heavy ion beam mutation, which provided a good theoretical basis and application basis for further development of this strain.
  • [1]
    ISLAM R, HOSSAIN M N, ALAM M K, et al. Antibacterial activity of lactic acid bacteria and extraction of bacteriocin protein[J]. Advances in Bioscience and Biotechnology,2020,11(2):49−59. doi: 10.4236/abb.2020.112004
    [2]
    KAPRASOB R, KERDCHOECHUEN O, LAOHAKUNJIT N, et al. Fermentation-based biotransformation of bioactive phenolics and volatile compounds from cashew apple juice by select lactic acid bacteria[J]. Process Biochemistry,2017,59:141−149. doi: 10.1016/j.procbio.2017.05.019
    [3]
    DO T, BAO K T, TRAN T, et al. Decoding the capability of Lactobacillus plantarum W1 isolated from soybean whey in producing an exopolysaccharide[J]. ACS Omega,2020,5(51):33387−33394. doi: 10.1021/acsomega.0c05256
    [4]
    TAMBUR Z, MILJKOVI-SELIMOVI B, OPAI D, et al. Inhibitory effects of different medicinal plants on growth of some oral microbiome member[J]. Medycyna Weterynaryjna,2020,76(8):6433−2020. doi: 10.21521/mw.6433
    [5]
    SOPHATHA B, PIWAT S, TEANPAISAN R. Adhesion, anti-adhesion and aggregation properties relating to surface charges of selected Lactobacillus strains: Study in Caco-2 and H357 cells[J]. Archives of Microbiology,2020,202(6):1349−1357. doi: 10.1007/s00203-020-01846-7
    [6]
    SALEHIZADEH M, MODARRESSI M H, MOUSAVI S N, et al. Evaluation of lactic acid bacteria isolated from poultry feces as potential probiotic and its in vitro competitive activity against Salmonella typhimurium[C]//Veterinary Research Forum. Faculty of Veterinary Medicine, Urmia University, Urmia, Iran, 2020, 11(1): 67.
    [7]
    JACKMAN C M, DEANS K W, FORNEY L J, et al. Microdroplet co-cultivation and interaction characterization of human vaginal bacteria[J]. Integrative Biology,2019,11(3):69−78. doi: 10.1093/intbio/zyz006
    [8]
    ADEOSHUN F G, RUPPITSCH W, ALLERBERGER F, et al. Prevalence and antimicrobial properties of lactic acid bacteria in Nigerian women during the menstrual cycle[J]. Polish Journal of Microbiology,2019,68(2):203−209. doi: 10.33073/pjm-2019-020
    [9]
    KOSTELAC D, GERIĆ M, GAJSKI G, et al. Lactic acid bacteria isolated from equid milk and their extracellular metabolites show great probiotic properties and anti-inflammatory potential[J]. International Dairy Journal,2021,112:104828. doi: 10.1016/j.idairyj.2020.104828
    [10]
    MILJKOVIC M, MARINKOVIC P, NOVOVIC K, et al. AggLr, a novel aggregation factor in Lactococcus raffinolactis BGTRK10-1: Its role in surface adhesion[J]. Biofouling,2018,34(5−6):685−698.
    [11]
    陈舜华. 乳酸菌阴道胶囊对产后6周妇女特异性阴道炎防治作用的临床观察[J]. 家庭医药. 就医选药,2017(6):38−39. [CHEN S H. Clinical observation of Lactobacillus vaginalis capsule on prevention and treatment of specific vaginitis in women 6 weeks after delivery[J]. Family Medicine. Medical Selection of Medicine,2017(6):38−39.

    CHEN S H. Clinical observation of Lactobacillus vaginalis capsule on prevention and treatment of specific vaginitis in women 6 weeks after delivery[J]. Family Medicine. Medical Selection of Medicine, 2017(6): 38-39.
    [12]
    李艳欣. 犬源乳酸菌的筛选及其对番泻叶介导的犬腹泻的防治试验[D]. 长春: 吉林大学, 2018.

    LI Y X. Screening of canine Lactobacillus and its prevention and treatment of senna leaf mediated diarrhea in dogs[D]. Changchun: Jilin University, 2018.
    [13]
    BRON P A, MARCELLI B, MULDER J, et al. Renaissance of traditional DNA transfer strategies for improvement of industrial lactic acid bacteria[J]. Current Opinion in Biotechnology,2019,56:61−68. doi: 10.1016/j.copbio.2018.09.004
    [14]
    YOKOTA A. Breeding of useful microorganisms by mutation in energy metabolism[J]. Journal of the Agricultural Chemical Society of Japan,1997,71:9−14.
    [15]
    刘璐. 重离子束辐照恩拉霉素菌株的选育研究[D]. 北京: 中国科学院大学(中国科学院近代物理研究所), 2020.

    LIU L. Breeding of enramycin strain irradiated by heavy ion beam[D]. Beijing: University of Chinese Academy of Sciences (Institute of modern physics, Chinese Academy of Sciences), 2020.
    [16]
    LÜ Y, LI J, CHEN Z, et al. Species identification and mutation breeding of silicon-activating bacteria isolated from electrolytic manganese residue[J]. Environmental Science and Pollution Research,2020,28(2):1491−1501.
    [17]
    LIU K Y, FANG H, CUI F J, et al. ARTP mutation and adaptive laboratory evolution improve probiotic performance of Bacillus coagulans[J]. Applied Microbiology and Biotechnology,2020,104(14):6363−6373. doi: 10.1007/s00253-020-10703-y
    [18]
    WANG D, ZHANG T, YE H, et al. In vitro probiotic screening and evaluation of space-induced mutant Lactobacillus plantarum[J]. Food Science & Nutrition,2020,8(11):6031−6036.
    [19]
    GUO X, ZHANG M, GAO Y, et al. Repair characteristics and time-dependent effects in response to heavy-ion beam irradiation in Saccharomyces cerevisiae: A comparison with X-ray irradiation[J]. Applied Microbiology and Biotechnology,2020,104(4):4043−4057.
    [20]
    杨阳. 重离子诱变谷氨酸高产菌株选育及阿莫西林诱导发酵机理研究[D]. 兰州: 兰州理工大学, 2019.

    YANG Y. Screening of high yield glutamate strain induced by heavy ion and study on the mechanism of amoxicillin induced fermentation[D]. Lanzhou: Lanzhou University of Technology, 2019.
    [21]
    王志. 利用重离子碰撞研究原子核内核子的短程关联及高动量分布[D]. 南京: 南京大学, 2019.

    WANG Z. Study of short-range correlation and high momentum distribution of nucleons in nuclei by heavy ion collisions[D]. Nanjing: Nanjing University, 2019.
    [22]
    贾蓉, 苏锋涛, 胡步荣. 重离子的辐射生物效应及其在生命科学中的应用[J]. 生物技术通报,2018,34(1):67−78. [JIA R, SU F T, HU B R. Biological effects of heavy ions and their applications in life sciences[J]. Biotechnology Bulletin,2018,34(1):67−78. doi: 10.13560/j.cnki.biotech.bull.1985.2017-0735

    JIA R, SU F T, HU B R. Biological effects of heavy ions and their applications in life sciences[J]. Biotechnology Bulletin, 2018, 34(1): 67-78. doi: 10.13560/j.cnki.biotech.bull.1985.2017-0735
    [23]
    孙玲, 刘利平, 徐婉茹, 等. 物理诱变在药食用菌育种中的应用研究进展[J]. 安徽农业科学,2018,46(14):29−33, 153. [SUN L, LIU L P, XU W R, et al. Research progress in the application of physical mutagenesis in the breeding of medicinal edible fungi[J]. Journal of Anhui Agricultural Sciences,2018,46(14):29−33, 153. doi: 10.3969/j.issn.0517-6611.2018.14.009

    SUN L, LIU L P, XU W R, et al. Research progress in the application of physical mutagenesis in the breeding of medicinal edible fungi[J]. Journal of Anhui Agricultural Sciences, 2018, 46(14): 29-33, 153. doi: 10.3969/j.issn.0517-6611.2018.14.009
    [24]
    YANG Y N, LIU C L, WANG Y K, et al. Mutation effects of C2+ ion irradiation on the greasy Nitzschia sp.[J]. Mutation Research/Fundamental & Molecular Mechanisms of Mutagenesis,2013,751(4):24−28.
    [25]
    李垄清. 重离子诱变技术选育高产β-葡聚糖酵母菌株的研究[D]. 北京: 中国科学院大学(中国科学院近代物理研究所), 2017.

    LI L Q. Breeding of high-yield β-glucan yeast strains by heavy ion mutagenesis[D]. Beijing: University of Chinese Academy of Sciences (Institute of Modern Physics, Chinese Academy of Sciences), 2017.
    [26]
    WANG S, BO Y, CHEN J, et al. Effects of heavy-ion beam irradiation on avermectin B1a and its analogues production by Streptomyces avermitilis[J]. Engineering in Life Sciences,2018,18(10):711−720. doi: 10.1002/elsc.201800094
    [27]
    JIANG A, HU W, LI W, et al. Enhanced production of l‐lactic acid by Lactobacillus thermophilus SRZ50 mutant generated by high-linear energy transfer heavy ion mutagenesis[J]. Engineering in Life Sciences,2018,18(9):626−634. doi: 10.1002/elsc.201800052
    [28]
    缪建顺, 曹国珍, 张苗苗, 等. 重离子束诱变选育谷氨酸高产菌株[J]. 辐射研究与辐射工艺学报,2015,33(5):39−45. [MIAO J S, CAO G Z, ZHANG M M, et al. Breeding of high glutamic acid producing strain by heavy ion beam mutation[J]. Journal of Radiation Research and Radiation Processing Technology,2015,33(5):39−45. doi: 10.11889/j.1000-3436.2015.rrj.33.050401

    MIAO J S, CAO G Z, ZHANG M M, et al. Breeding of high glutamic acid producing strain by heavy ion beam mutation[J]. Journal of Radiation Research and Radiation Processing Technology, 2015, 33(5): 39-45 doi: 10.11889/j.1000-3436.2015.rrj.33.050401
    [29]
    DIANA R M, ERICA K, GONZALO M C, et al. Growth of lactic acid bacteria in milk phospholipids enhances their adhesion to Caco-2 cells[J]. Journal of Dairy Science,2016,103(9):7707−7718.
    [30]
    张金露, 吴涛, 唐艳. 生物传感仪-除NH4+法测定发酵液中L-谷氨酸含量的研究[J]. 中国食品添加剂,2021,32(1):86−91. [ZHANG J L, WU T, TANG Y. Study on determination of L-glutamic acid by biosensor removing NH4+ method[J]. China Food Additives,2021,32(1):86−91.

    ZHANG J L, WU T, TANG Y. Study on determination of L-glutamic acid by biosensor removing NH4+ method[J]. China Food Additives, 2021, 32(1): 86-91.
    [31]
    王雨辰. 重离子辐照诱变选育高产酸乳酸菌株及其发酵条件优化研究[D]. 兰州: 甘肃农业大学, 2017

    WANG Y C. Breeding of lactic acid producing strain by heavy ion irradiation and optimization of fermentation conditions[D]. Lanzhou: Gansu Agricultural University, 2017.
    [32]
    陈积红, 胡伟, 李文建, 等. 重离子束12C6+累进辐照诱变柠檬酸菌株选育研究[J]. 原子核物理评论,2013,30(4):483−487. [CHEN J H, HU W, LI W J, et al. Breeding of citric acid strain induced by heavy ion beam 12C6+ progressive irradiation[J]. Nuclear Physics Review,2013,30(4):483−487. doi: 10.11804/NuclPhysRev.30.04.483

    CHEN J H, HU W, LI W J, et al. Breeding of citric acid strain induced by heavy ion beam 12C6+ progressive irradiation[J]. Nuclear Physics Review, 2013, 30(4): 483-487. doi: 10.11804/NuclPhysRev.30.04.483
    [33]
    ABOULOIFA H, ROKNI Y, BELLAOUCHI R, et al. Characterization of probiotic properties of antifungal Lactobacillus strains isolated from traditional fermenting green olives[J]. Probiotics & Antimicrobial Proteins,2019,12(2):683−696.
    [34]
    佘之蕴, 黄宝莹, 刘海卿, 等. 牛津杯法测定食品添加剂对五种益生菌的抑菌活力[J]. 食品工业,2016,37(1):171−174. [SHE Z Y, HUANG B Y, LIU H Q, et al. Determination of the antibacterial activity of food additives against five probiotics by Oxford cup method[J]. Food Industry,2016,37(1):171−174.

    SHE Z Y, HUANG B Y, LIU H Q, et al. Determination of the antibacterial activity of food additives against five probiotics by Oxford cup method[J]. Food Industry, 2016, 37(1): 171-174.
    [35]
    吴庆华, 陈积红, 张珍, 等. X射线对嗜热乳杆菌产L-乳酸的选育研究[J]. 食品工业科技,2015,36(3):116−118,127. [WU Q H, CHEN J H, ZHANG Z, et al. Breeding of L-lactic acid produced by Lactobacillus thermophilus by X-ray[J]. Science and Technology of Food Industry,2015,36(3):116−118,127. doi: 10.13386/j.issn1002-0306.2015.03.015

    WU Q H, CHEN J H, ZHANG Z, et al. Breeding of L-lactic acid produced by Lactobacillus thermophilus by X-ray[J]. Science and Technology of Food Industry, 2015, 36(3): 116-118, 127. doi: 10.13386/j.issn1002-0306.2015.03.015
    [36]
    徐颖, 贺黎, 吕嘉枥, 等. 富硒鼠李糖乳杆菌稳定性及其冻干保护剂研究[J]. 中国食品学报,2020,20(9):102−108. [XU Y, HE L, LÜ J Z, et al. Study on the stability of selenium enriched Lactobacillus rhamnosus and its freeze-drying protectant[J]. Journal of Chinese Institute of Food Science and Technology,2020,20(9):102−108. doi: 10.16429/j.1009-7848.2020.09.013

    XU Y, HE L, LÜ J Z, et al. Study on the stability of selenium enriched Lactobacillus rhamnosus and its freeze-drying protectant[J]. Journal of Chinese Institute of Food Science and Technology, 2020, 20(9): 102-108. doi: 10.16429/j.1009-7848.2020.09.013
    [37]
    罗素贤, 叶昱, 周信荣, 等. 一株鼠李糖乳杆菌的培养条件优化研究[J]. 江西农业大学学报,2018,40(2):365−370. [LUO S X, YE Y, ZHOU X R, et al. Optimization of culture conditions of a Lactobacillus rhamnosus[J]. Journal of Jiangxi Agricultural University,2018,40(2):365−370. doi: 10.13836/j.jjau.2018048

    LUO S X, YE Y, ZHOU X R, et al. Optimization of culture conditions of a Lactobacillus rhamnosus[J]. Journal of Jiangxi Agricultural University, 2018, 40(2): 365-370. doi: 10.13836/j.jjau.2018048
    [38]
    王雨辰, 王曙阳, 董妙音, 等. 重离子束辐照选育高产植物乳酸菌[J]. 辐射研究与辐射工艺学报,2017,35(1):52−58. [WANG Y C, WANG S Y, DONG M Y, et al. Breeding of high yield plant lactic acid bacteria by heavy ion beam irradiation[J]. Journal of Radiation Research and Radiation Processing Technology,2017,35(1):52−58. doi: 10.11889/j.1000-3436.2017.rrj.35.010401

    WANG Y C, WANG S Y, DONG M Y, et al. Breeding of high yield plant lactic acid bacteria by heavy ion beam irradiation[J]. Journal of Radiation Research and Radiation Processing Technology, 2017, 35(1): 52-58. doi: 10.11889/j.1000-3436.2017.rrj.35.010401
    [39]
    都雯玥. 重离子辐照并筛选截短侧耳素高产菌株的研究[D]. 兰州: 兰州理工大学, 2016.

    DU W Y. Study on the high yield strains of truncated lateral otoxin by heavy ion irradiation[D]. Lanzhou: Lanzhou University of Technology, 2016.
    [40]
    LIU J, QI Z, HUANG Q, et al. Study of energetic-particle-irradiation induced biological effect on Rhizopus oryzae through synchrotron-FTIR micro-spectroscopy[J]. Journal of Molecular Structure,2013,1031:1−8. doi: 10.1016/j.molstruc.2012.07.025
    [41]
    蔡聪, 姜婷, 郑兆娟, 等. 等离子体诱变凝结芽孢杆菌提高木糖利用能力高产 L-乳酸[J]. 食品科学,2014,35(1):125−129. [CAI C, JIANG T, ZHENG Z J, et al. Improved xylose utilization of Bacillus coagulans by atmospheric and room temperature plasma mutation for production of lactic acid[J]. Food Science,2014,35(1):125−129. doi: 10.7506/spkx1002-6630-201401024

    CAI C, JIANG T, ZHENG Z J, et al. Improved xylose utilization of Bacillus coagulans by atmospheric and room temperature plasma mutation for production of lactic acid[J]. Food Science, 2014, 35(1): 125-129. doi: 10.7506/spkx1002-6630-201401024
    [42]
    杨佩斯. ARTP诱变黑曲霉絮凝菌株及应用研究[D]. 贵阳: 贵州大学, 2020.

    YANG P S. Mutation of Aspergillus niger flocculating strain by ARTP and its application[D]. Guiyang: Guizhou University, 2020.
    [43]
    杨然, 范光森, 郦金龙, 等. 重组毕赤酵母高产木聚糖酶菌株筛选及发酵条件优化[J]. 中国食品学报,2017,17(12):95−104. [YANG R, FAN G S, LI J L, et al. Screening of recombinant Pichia pastoris strains with high xylanase production and optimization of fermentation conditions[J]. Journal of Chinese Institute of Food Science and Technology,2017,17(12):95−104. doi: 10.16429/j.1009-7848.2017.12.013

    YANG R, FAN G S, LI J L, et al. Screening of recombinant Pichia pastoris strains with high xylanase production and optimization of fermentation conditions[J]. Journal of Chinese Institute of Food Science and Technology, 2017, 17(12): 95-104. doi: 10.16429/j.1009-7848.2017.12.013

Catalog

    Article Metrics

    Article views (128) PDF downloads (9) Cited by()

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return