Abstract
BackgroundMenaquinones 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.ResultsMenaquinones 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.ConclusionThe LCM method is a simple, rapid and efficient technique for the extraction and identification of menaquinones from Actinomycetes.
Highlights
Menaquinones are constituents of prokaryote cell membranes where they play important functions during electron transport
For the lysozyme-chloroform-methanol (LCM) method proposed in this study, wet cells can be directly used for MK extraction
The Collins method requires hours or even an overnight period to acquire freeze-dried cells and an overnight extraction is needed to obtain the crude extract of MKs
Summary
Menaquinones are constituents of prokaryote cell membranes where they play important functions during electron transport. The most widely used methods to determine menaquinones are based on freeze-dried cells. MK extraction and determination methods for Actinomycetes were described by Collins et al in a paper that currently has more than 2000 citations [4,5,6,7,8]. A respiratory lipoquinone extraction method was reported in 2011 [9], while a screening method for MKproducing strains was reported in 2020 [10]. All of these MK extraction methods use freeze-dried cells. As the most widely used method, we applied the Collins et al (1977) methodology to test the MKs from Actinomycetes strains in our lab. The respiratory quinones from Microcella putealis CV-2T, CV-40 and AC-30 have been detected at low concentrations
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