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- PROTOCOL] Applications of different solvents and conditions for differential extraction of lipopolysaccharide in Gram-negative bacteria
- Mai Phuong Nguyen , Le Viet Ha Tran , Hyun Namgoong , Yong-Hak Kim
- J. Microbiol. 2019;57(8):644-654. Published online May 23, 2019
- DOI: https://doi.org/10.1007/s12275-019-9116-5
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- 6 Web of Science
- 6 Crossref
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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. -
Citations
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