A rapid and efficient method for the extraction and identification of menaquinones from Actinomycetes in wet biomass

BMC Microbiol. 2021 Jun 9;21(1):175. doi: 10.1186/s12866-021-02240-z.

Abstract

Background: Menaquinones are constituents of prokaryote cell membranes where they play important functions during electron transport. Menaquinone profiles are strongly recommended for species classification when proposing a new Actinomycetes taxon. Presently, the most widely used methods to determine menaquinones are based on freeze-dried cells. Taxonomic research in our lab has revealed that menaquinone concentrations are low for some species of the genus Microbacterium, leading to difficulties in identifying menaquinones.

Results: Menaquinones extracted using the novel lysozyme-chloroform-methanol (LCM) method were comparable in quality to those obtained using the Collins method, the most widely used method. All tested strains extracted via the LCM method showed higher concentrations of menaquinones than those extracted via the Collins method. For some Microbacterium strains, the LCM method exhibited higher sensitivity than the Collins method, and more trace menaquinones were detected with the LCM method than the Collins method. In addition, LCM method is faster than the Collins method because it uses wet cells.

Conclusion: The LCM method is a simple, rapid and efficient technique for the extraction and identification of menaquinones from Actinomycetes.

Keywords: Actinomycetes; Menaquinone analysis; Menaquinone extraction; Menaquinone identification.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actinobacteria / chemistry*
  • Actinobacteria / growth & development
  • Actinobacteria / metabolism
  • Biomass
  • Chemical Fractionation / methods*
  • Chloroform / chemistry
  • Hypertonic Solutions / chemistry
  • Methanol / chemistry
  • Vitamin K 2 / chemistry
  • Vitamin K 2 / isolation & purification*
  • Vitamin K 2 / metabolism

Substances

  • Collins' solution
  • Hypertonic Solutions
  • Vitamin K 2
  • Chloroform
  • Methanol