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An improved sampling protocol for analysis of intracellular metabolites in Mortierella alpina

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Abstract

Sampling of intracellular metabolites in Mortierella alpina was investigated as part of a metabolomics study. After comparison of four sampling protocols, rapid filtration of the culture using a laboratory-made nylon filter and absorbent gauze under normal pressure followed by quenching in liquid N2 and grinding (the improved protocol) was the most effective. Rapid filtration under normal pressure decreased intracellular metabolites leakage and subsequent grinding of cells contributed to intracellular metabolites extraction. The above quenching method together with 75 % (v/v) ethanol, buffered with 60 mM HEPES, at 80 °C for 3 min is therefore suitable for sampling intracellular metabolites in M. alpina.

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Acknowledgments

This work was financially supported by the National Basic Research Program of China (No. 2011CBA00802), the National Natural Science Foundation of China (No. 21176124) and the Key Program (No. 20936002), the Key Technology Support Program of China (No. 2011BAD23B03), and the Priority Academic Program Development of Jiangsu Higher Education Institutions. H. Huang was supported by the Fok Ying Tung Education Foundation (No. 123014), and the Program for New Century Excellent Talents in University from the Ministry of Education of China (No. NCET-09-0157).

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Authors and Affiliations

  1. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, No. 5 Xinmofan Road, Nanjing, 210009, People’s Republic of China

    Xin Liu, Xiaojun Ji, Ninghua Fu & He Huang

  2. College of Sciences, Nanjing University of Technology, No. 5 Xinmofan Road, Nanjing, 210009, People’s Republic of China

    Hongman Zhang, Hongbo Zheng & Xin Zhang

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  1. Xin Liu
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  2. Hongman Zhang
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  3. Xiaojun Ji
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  4. Hongbo Zheng
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  5. Xin Zhang
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  6. Ninghua Fu
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  7. He Huang
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Correspondence to He Huang.

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Liu, X., Zhang, H., Ji, X. et al. An improved sampling protocol for analysis of intracellular metabolites in Mortierella alpina . Biotechnol Lett 34, 2275–2282 (2012). https://doi.org/10.1007/s10529-012-1030-4

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  • DOI: https://doi.org/10.1007/s10529-012-1030-4

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