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Mg, K-containing microparticle: A possible active principle of a culture extract produced by a microbial consortium
Toru Higashinakagawa1,2, Haruhisa Kikuchi3, Hidekazu Kuwayama4
1International Center for Molecular, Cellular and Immunological Research, Tokyo Women's Medical University, Tokyo, Japan.
Abstract:
A synthetic microbial consortium called Effective Microorganisms (EM) consists mainly of photosynthetic bacteria, lactic acid bacteria and yeast. Various effects of EM∙XGOLD, a health drink produced by EM, on life cycle of Dictyostelium discoideum were described previously. Here, we report our attempt to identify the active principle, termed EMF, that brought about the observed effects. Throughout the purification processes, the presence of the active principle was monitored by promoted fruiting body formation. By liquid-liquid separation the activity was recovered in aqueous phase, which, after concentration, was further subjected to reverse-phase column chromatography. No activity was detected in any eluant, while almost all the activity was recovered in residual insoluble material. The application of conventional organic chemistry procedures to the residual fraction did not lead to any informative results. Acid treatment of the insoluble material produced air bubbles, suggesting it to be composed of some inorganic carbonate. Viewed under scanning electronmicroscope, the residue revealed spherical particles of μm size range. Energy Dispersive X-ray (EDX) Spectroscopy pointed to the existence, on the surface of the particles, of magnesium and, to a certain extent, of potassium. In separate experiments, acid treatment and alkali neutralization of EM∙XGOLD completely wiped out the stimulatory activity of fruiting body formation. These lines of evidence indicate these Mg, K-containing microparticles to be an active principle of EM culture extract. How these particles exert their effect is currently under intensive investigation.
Insights
Researchers identified microparticles containing magnesium and potassium as the active principle in EM∙XGOLD, a health drink derived from Effective Microorganisms (EM). These particles stimulate fruiting body formation in Dictyostelium discoideum.
Area of Science:
- Microbiology
- Biochemistry
- Developmental Biology
Background:
- Effective Microorganisms (EM) is a synthetic microbial consortium including photosynthetic bacteria, lactic acid bacteria, and yeast.
- EM∙XGOLD, a health drink produced by EM, has demonstrated effects on the life cycle of Dictyostelium discoideum.
- The active principle responsible for these effects, termed EMF, remained unidentified.
Purpose of the Study:
- To identify the active principle (EMF) in EM∙XGOLD responsible for promoting fruiting body formation in Dictyostelium discoideum.
- To characterize the chemical nature of the active principle.
Main Methods:
- Purification of the active principle using liquid-liquid separation and reverse-phase column chromatography.
- Analysis of the insoluble residual fraction using scanning electron microscopy (SEM) and Energy Dispersive X-ray (EDX) Spectroscopy.
- Assessment of activity by monitoring promoted fruiting body formation.
- Chemical treatments (acid and alkali) to evaluate the stability and nature of the active principle.
Main Results:
- The activity was localized to an insoluble material after purification.
- SEM revealed μm-sized spherical particles in the residue.
- EDX spectroscopy identified magnesium and potassium on the surface of these microparticles.
- Acid treatment of EM∙XGOLD and neutralization of alkali treatments abolished the stimulatory activity.
Conclusions:
- Mg, K-containing microparticles are identified as the active principle in EM culture extract responsible for promoting fruiting body formation.
- The physical and chemical properties of these microparticles suggest an inorganic carbonate composition.
- Further investigation is ongoing to elucidate the mechanism of action of these microparticles.
