[1]罗顺,刘冰,张金龙,等.混凝土自修复菌Bacillus sp.芽孢生产和萌发条件的优化[J].深圳大学学报理工版,2020,37(1):25-32.[doi:10.3724/SP.J.1249.2020.01025]
　LUO Shun,LIU Bing,ZHANG Jinlong,et al.Optimization of sporulation and germination of self-healing bacterium Bacillus sp. for concrete crack[J].Journal of Shenzhen University Science and Engineering,2020,37(1):25-32.[doi:10.3724/SP.J.1249.2020.01025]
点击复制

混凝土自修复菌Bacillus sp.芽孢生产和萌发条件的优化()
分享到：

《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第37卷
期数:
2020年第1期
页码:
25-32
栏目:
生物工程
出版日期:
2020-01-08

文章信息/Info

Title:
Optimization of sporulation and germination of self-healing bacterium Bacillus sp. for concrete crack
文章编号:
202001004
作者:
罗顺1; 刘冰2; 张金龙1; 李秋伟1; 邓旭1
1）深圳大学生命与海洋科学学院，广东深圳 518055
2）深圳信息职业技术学院交通与环境学院，广东深圳 518172
Author(s):
LUO Shun1;  LIU Bing2;  ZHANG Jinlong1;  LI Qiuwei1;  and DENG Xu1
1) College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, Guangdong Province, P.R.China
2) School of Traffic and Environment, Shenzhen Institute of Information Technology, Shenzhen 518172, Guangdong Province, P.R.China
关键词:
微生物学; 自修复; 枯草芽孢菌属; 芽孢生成; 萌发; 矿化
Keywords:
microbiology;  self-healing;  Bacillus;  sporulation;  germination;  mineralization
分类号:
Q819;Q939.99
DOI:
10.3724/SP.J.1249.2020.01025
文献标志码:
A
摘要:
对一株混凝土自修复菌Bacillus sp.B8芽孢生产条件进行优化，研究芽孢萌发的影响因素.芽孢生产的单因素实验表明，最适接种量为1×107 mL-1；最适碳源和氮源分别为淀粉和酵母粉，相应的最适质量浓度分别为0.5 g/L和2 g/L；一定质量浓度的Mg2+(0.24 g/L)和Mn2+(4.8 mg/L)有利于芽孢生成. 在单因素实验的基础上利用正交实验对各影响因子进行了优化，结果表明，接种量和酵母粉浓度是影响芽孢产量的显著因子，在接种量、淀粉、酵母粉、Mg2+和Mn2+浓度分别为1×107 mL-1、5、5、0.48和4.8×10-3 g/L时，芽孢产量达到1.54×109 mL-1，比优化前提高150倍. 此外，芽孢萌发实验表明，芽孢萌发时间约为4 h；最适萌发剂为肌苷，最适质量浓度为2 g/L，最适pH值为10；金属离子Na+能促进芽孢萌发，但Ca2+对芽孢的萌发有严重的抑制作用.在实验中首次发现一定质量浓度的Na+能消除Ca2+的抑制作用，最适质量浓度为24 g/L.
Abstract:
The single factor experimental results of spore production of a self-healing bacterial strain, Bacillus sp.B8, show that the optimal inoculation volume is 1×107 mL-1. The suitable carbon and nitrogen source are starch and yeast extract and the corresponding optimal concentrations being 0.5 g/L and 2 g/L, respectively. The presence of Mg2+ (0.24 g/L) and Mn2+ (4.8 mg/L) is beneficial for spore formation. Furthermore, based on the above results, orthogonal experiment is carried out to optimize the influential factors. The result shows that the inoculation volume and yeast extract concentration are the significant factors affecting spore yield. Under the optimized condition in which the concentrations of inoculation cells, starch, yeast extract, Mg2+ and Mn2+ are 1×107 mL-1, 5, 5, 0.48 and 4.8×10-3 g/L, respectively, the final spore concentration reaches up to 1.54×109 mL-1, showing more than 150 times enhancement. On the other hand, the results of spore germination indicate that spores need about 4 h to complete germination process. The most suitable germinant is inosine, and the optimal concentration is 2 g/L. The suitable pH is 10. Na+ could promote the germination of spores, whereas Ca2+ shows a severe inhibition on the germination of spores. However, we find, for the first time, that the presence of Na+ at a certain concentration (the optimal concentration is 24 g/L) can alleviate the inhibition of Ca2+ on spore germination to a large extent.

参考文献/References:

［1］ BOQUET E, BORONAT A, RAMOS A. Production of calcite (calcium carbonate) crystals by soil bacteria is a general phenomenon［J］. Nature, 1973,246(5434): 527-529.
［2］ GOLLAPUDI U K, KNUTSON C L, BANG S S, et al. A new method for controlling leaching through permeable channels［J］. Chemospher, 1995,30(4): 695-705.
［3］ WANG J, ERSAN Y C, BOON N, et al. Application of microorganisms in concrete: a promising sustainable strategy to improve concrete durability［J］. Applied Microbiology and Biotechnology, 2016,100(7):2993-3007.
［4］ BUNDUR Z B, AMIRI A, ERSAN Y C, et al. Impact of air entraining admixtures on biogenic calcium carbonate precipitation and bacterial viability［J］. Cement and Concrete Research, 2017,98:44-49.
［5］ NAIN N, SURABHI R, YATHISH N V, et al. Enhancement in strength parameters of concrete by application of Bacillus bacteria［J］. Construction and Building Materials, 2019,202:904-908.
［6］ LORS C, DUCASSE-LAPEYRUSSE J, GAGN R, et al. Microbiologically induced calcium carbonate precipitation to repair microcracks remaining after autogenous healing of mortars［J］. Construction and Building Materials, 2017,141:461-469.
［7］ 严涛, 朱建国, 姜甜, 等. 一株凝结芽孢杆菌的分离筛选及产孢条件优化［J］. 微生物学通报, 2018,45(2):238-249.
YAN Tao, ZHU Jianguo, JIANG Tian, et al. Isolation and optimization on spore-forming conditions of Bacillus coagulans［J］. Microbiology China, 2018,45(2):238- 249.(in Chinese)
［8］ 左肖肖, 陆利霞, 李壹, 等. 驱油枯草芽孢杆菌产芽孢条件优化［J］. 科学技术与工程, 2017,17(16):28-34.
ZUO Xiaoxiao, LU Lixia, LI Yi, et al. The Formation and germination of the spores of Bacillus subtilis［J］. Science Technology and Engineering, 2017,17(16): 28-34.(in Chinese)
［9］ 吴迎奔, 陈薇, 贺月林, 等. 侧孢芽孢杆菌 C5发酵条件及培养基的优化［J］. 湖南农业科学, 2016(9):4-6.
WU Yingben, CHEN Wei, HE Yuelin, et al. Optimizing fermentation condition for Bacillus laterosporus C5［J］. Hunan Agricultural Sciences, 2016(9):4-6.(in Chinese)
［10］ 冯鑫, 何承云, 徐茜茜, 等. 培养基成分对酸土脂环酸芽孢杆菌生长及芽孢形成的影响［J］. 食品工业科技, 2018,39(19):84-89.
FENG Xin, HE Chengyun, XU Qianqian, et al. Effect of medium component on growth and spore formation of Alicyclobacillus acidoterrestris［J］. Science and Technology of Food Industry, 2018,39(19):84-89.(in Chinese)
［11］ 董佩佩, 汪祥燕, 刘元香, 等. 一株凝结芽孢杆菌(Bacillus coagulans)发酵培养基的优化［J］. 中国酿造, 2018,37(4):28-32.
DONG Peipei, WANG Xiangyan, LIU Yuanxiang, et al. Optimization of fermentation medium of Bacillus coagulans［J］. China Brewing, 2018,37(4):28-32.(in Chinese)
［12］ SHARMA T K, ALAZHARI M, HEATH A, et al. Alkaliphilic Bacillus species show potential application in concrete crack repair by virtue of rapid spore production and germination then extracellular calcite formation［J］. Journal of Applied Microbiology, 2017,122(5):1233-1244.
［13］ 王一晓, 张海红, 章中. 理化因素诱导芽孢萌发研究进展［J］. 农业科学研究, 2016,37(1):58-64.
WANG Yixiao, ZHANG Haihong, ZHANG Zhong. Research progress on spore germination induced by physical and chemical factors［J］. Journal of Agricultural Sciences, 2016,37(1):58-64.(in Chinese)
［14］ 程文凤, 彭慧, 刘冰, 等. 混凝土修复功能菌Bacillus cohnii DSM6307芽孢萌发条件探究［J］. 科技通报, 2016,32(5):219-223.
CHENG Wenfeng, PENG Hui, LIU Bing, et al. Study on the condition of spore germination of concrete self-healing bacterium Bacillus cohnii DSM6307［J］. Bulletin of Science and Technology, 2016,32(5):219- 223.(in Chinese)
［15］ GOUNINA-ALLOUANE R, BROUSSOLLE V, CARLIN F. Influence of the sporulation temperature on the impact of the nutrients inosine and l-alanine on Bacillus cereus spore germination［J］. Food Microbiology, 2008,25(1):202-206.
［16］ VERCAMMEN A, VIVIJS B, LURQUIN I, et al. Germination and inactivation of Bacillus coagulans and Alicyclobacillus acidoterrestris spores by high hydrostatic pressure treatment in buffer and tomato sauce［J］. International Journal of Food Microbiology, 2012,152(3):162-167.
［17］ SETLOW B, SETLOW P. Measurements of the pH within dormant and germinated bacterial spores［J］. Biochemistry, 1980,77(5):2474-2476.
［18］ SOUTHWORTH T W, GUFFANTI A A, MOIR A, et al. GerN, an endospore germination protein of Bacillus cereus, is an Na+/H+-K+ antiporter［J］. Journal of Bacteriology, 2001,183(20):5896-5903.

相似文献/References:

[1]倪卓,杜学晓,邢锋,等.表面活性剂对自修复环氧树脂微胶囊的影响[J].深圳大学学报理工版,2008,25(4):351.
　NI Zhuo,DU Xue-xiao,XING Feng,et al.Effects of surfactant on MUF encapsulated epoxy in self-healing[J].Journal of Shenzhen University Science and Engineering,2008,25(1):351.
[2]邢锋,倪卓,汤皎宁,等.自修复混凝土系统的研究进展[J].深圳大学学报理工版,2013,30(No.5(441-550)):486.[doi:10.3724/SP.J.1249.2013.05486]
　Xing Feng,Ni Zhuo,Tang Jiaoning,et al.State-of-the-art for self-healing concrete[J].Journal of Shenzhen University Science and Engineering,2013,30(1):486.[doi:10.3724/SP.J.1249.2013.05486]
[3]罗凯,刘丹丹,周丽蓉,等.黑荆树提取物对铜绿微囊藻的化感抑制效应[J].深圳大学学报理工版,2014,31(2):205.[doi:10.3724/SP.J.1249.2014.02205]
　Luo Kai,Liu Dandan,Zhou Lirong,et al.Allelopathic inhibitory effect of the Acacia mearnsii extracts on Microcystis Aeruginosa[J].Journal of Shenzhen University Science and Engineering,2014,31(1):205.[doi:10.3724/SP.J.1249.2014.02205]
[4]胡浪,刘烨蓉,许伟钊,等.集胞藻缺氮驯化株的代谢组学分析[J].深圳大学学报理工版,2019,36(5):570.[doi:10.3724/SP.J.1249.2019.05570]
　HU Lang,LIU Yerong,XU Weizhao,et al.Metabonomics analysis of nitrogen deficiency-generated strains of Synechocystis sp.[J].Journal of Shenzhen University Science and Engineering,2019,36(1):570.[doi:10.3724/SP.J.1249.2019.05570]

备注/Memo

备注/Memo:
Received：2019-05-20；Accepted：2019-09-19
Foundation：National Natural Science Foundation of China (51578339); Project of Department of Education of Guangdong Province (2017GKTSCX064)
Corresponding author：Professor DENG Xu. E-mail: dengxu@szu.edu.cn
Citation：LUO Shun，LIU Bing，ZHANG Jinlong，et al. Optimization of sporulation and germination of self-healing bacterium Bacillus sp. for concrete crack［J］. Journal of Shenzhen University Science and Engineering, 2020, 37(1): 25-32.（in Chinese）
基金项目：国家自然科学基金资助项目(51578339);广东省高等学校科技创新资助项目(2017GKTSCX064)
作者简介：罗顺(1995—)，男，深圳大学硕士研究生.研究方向：混凝土的微生物自修复. E-mail：luoshun2017@email.szu.edu.cn
引文：罗顺，刘冰，张金龙，等. 混凝土自修复菌Bacillus sp.芽孢生产和萌发条件的优化［J］. 深圳大学学报理工版，2020，37(1)：25-32.
更新日期/Last Update: 2020-01-30