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Applications of different solvents and conditions for differential extraction of lipopolysaccharide in Gram-negative bacteria

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Abstract

Lipopolysaccharide (LPS) is one of the major components in the outer membrane of Gram-negative bacteria. However, its heterogeneity and variability in different bacteria and differentiation conditions make it difficult to extract all of the structural variants. We designed a solution to improve quality and biological activity of LPS extracted from various bacteria with different types of LPS, as compared to conventional methods. We introduced a quality index as a simple measure of LPS purity in terms of a degree of polysaccharide content detected by absorbance at 204 nm. Further experiments using gel electrophoresis, endotoxin test, and macrophage activation test were performed to evaluate the performance and reliability of a proposed ‘T-sol’ method and the biological effectiveness and character of the LPS products. We presented that the T-sol method had differential effects on extraction of a RAW 264.7 cell-activating LPS, which was effective in the macrophage activation with similar effects in stimulating the production of TNF-alpha. In conclusion, the T-sol method provides a simple way to improve quality and biological activity of LPS with high yield.

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Acknowledgements

This study was supported by the National Research Foundation of Korea funded by the Ministry of Science, ICT and Future Planning (Grant number 2016R1A2B2014493 to YHK).

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  1. These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology, Daegu Catholic University School of Medicine, Daegu, 42472, Republic of Korea

    Mai Phuong Nguyen, Le Viet Ha Tran & Yong-Hak Kim

  2. Kolon R&D Division, Seoul, 07793, Republic of Korea

    Hyun Namgoong

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  1. Mai Phuong Nguyen
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Correspondence to Yong-Hak Kim.

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Nguyen, M.P., Tran, L.V.H., Namgoong, H. et al. Applications of different solvents and conditions for differential extraction of lipopolysaccharide in Gram-negative bacteria. J Microbiol. 57, 644–654 (2019). https://doi.org/10.1007/s12275-019-9116-5

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