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KMID : 0880220190570080644
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Journal of Microbiology
2019 Volume.57 No. 8 p.644 ~ p.654
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Applications of different solvents and conditions for differential extraction of lipopolysaccharide in Gram-negative bacteria
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Nguyen Mai Phuong
Tran Le Viet Ha
Namgoong Hyun
Kim Yong-Hak
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Abstract
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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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KEYWORD
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Gram-negative bacteria, lipopolysaccharide (LPS), quality index, effective does (ED50), macrophage activation
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