Han Pei-pei, Jia Shi-ru, Sun Ying, Tan Zhi-lei, Zhong Cheng, Dai Yu-jie, Tan Ning, Shen Shi-gang
Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, People's Republic of China.
World J Microbiol Biotechnol. 2014 Sep;30(9):2407-18. doi: 10.1007/s11274-014-1666-4. Epub 2014 May 16.
The application of antibiotic treatment with assistance of metabolomic approach in axenic isolation of cyanobacterium Nostoc flagelliforme was investigated. Seven antibiotics were tested at 1-100 mg L(-1), and order of tolerance of N. flagelliforme cells was obtained as kanamycin > ampicillin, tetracycline > chloromycetin, gentamicin > spectinomycin > streptomycin. Four antibiotics were selected based on differences in antibiotic sensitivity of N. flagelliforme and associated bacteria, and their effects on N. flagelliforme cells including the changes of metabolic activity with antibiotics and the metabolic recovery after removal were assessed by a metabolomic approach based on gas chromatography-mass spectrometry combined with multivariate analysis. The results showed that antibiotic treatment had affected cell metabolism as antibiotics treated cells were metabolically distinct from control cells, but the metabolic activity would be recovered via eliminating antibiotics and the sequence of metabolic recovery time needed was spectinomycin, gentamicin > ampicillin > kanamycin. The procedures of antibiotic treatment have been accordingly optimized as a consecutive treatment starting with spectinomycin, then gentamicin, ampicillin and lastly kanamycin, and proved to be highly effective in eliminating the bacteria as examined by agar plating method and light microscope examination. Our work presented a strategy to obtain axenic culture of N. flagelliforme and provided a method for evaluating and optimizing cyanobacteria purification process through diagnosing target species cellular state.
研究了代谢组学方法辅助抗生素处理在丝状念珠藻无菌分离中的应用。测试了7种抗生素，浓度为1 - 100 mg L(-1)，得出丝状念珠藻细胞对这些抗生素的耐受顺序为卡那霉素 > 氨苄青霉素、四环素 > 氯霉素、庆大霉素 > 壮观霉素 > 链霉素。根据丝状念珠藻与相关细菌对抗生素敏感性的差异，选择了4种抗生素，并基于气相色谱 - 质谱联用结合多变量分析的代谢组学方法，评估了它们对丝状念珠藻细胞的影响，包括抗生素处理后细胞代谢活性的变化以及去除抗生素后的代谢恢复情况。结果表明，抗生素处理影响了细胞代谢，因为经抗生素处理的细胞在代谢上与对照细胞不同，但通过去除抗生素，代谢活性会恢复，所需的代谢恢复时间顺序为壮观霉素、庆大霉素 > 氨苄青霉素 > 卡那霉素。抗生素处理程序相应地优化为从壮观霉素开始，然后是庆大霉素、氨苄青霉素，最后是卡那霉素的连续处理，通过琼脂平板法和光学显微镜检查证明，该方法在消除细菌方面非常有效。我们的工作提出了一种获得丝状念珠藻无菌培养物的策略，并提供了一种通过诊断目标物种细胞状态来评估和优化蓝藻纯化过程的方法。
