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Respiratory Quinones Analysis

Bacterial respiratory quinones in environmental samples is regarded as the


sensitive indicators of aerobic metabolism and anaerobic metabolism in
bacterial populations. In many areas of microbial ecology, it’s quite useful to
improve the ability of measuring the relative abundance of aerobic and
anaerobic metabolism in environmental samples. For instance, this measure
can provide the predictive power when an environmental pollutant
experiences different biodegradation rates or pathways in aerobic
environments and anaerobic environments.
As mentioned before, the importance of aerobic and anaerobic metabolism in
environmental samples is realized nowadays. However, it’s impossible to
measure the redox potential or dissolved oxygen directly for many samples.
Further, it’s not accurate enough to enumerate aerobic microorganisms and
anaerobic microorganisms with plating or the other techniques. Bacterial
respiratory quinones are regarded as the biomarkers for the type of energy
metabolism found in microbial environments.
The common bacterial respiratory quinones could be divided into three
types: ubiquinones (UQ, l-methyl-2-isoprenyl-3,4-
dimethoxyparabenzoquinone), menaquinones (MK, 1-isoprenyl-2-methyl-
naphthoquinone) and desmethylmenaquinones (DMK, 1-
isoprenylnaphthoquinone). All these three types of quinones belong to a
homologous series of compounds. The main difference among these
compounds is the number of isoprene units in the side chain. Further, the type
of quinone and the length of the side chain varies depending on the type of
bacteria and the environmental conditions. Ubiquinones are the respiratory
quinones which could be found in eukaryotes and exist in some gram-
negative bacteria. Menaquinones could be found in gram-negative and gram-
positive bacteria and archaebacteria. However, desmethylmenaquinones are
not so widely distributed, which could only been found in some pathogenic
enterobacteria and streptococcus faecalis. Quinones could be widely used as
taxonomic markers for bacteria as the quinone content in a bacterial
community shift with changes in the availability of oxygen.
For the analysis of respiratory quinones, Creative Proteomics follow the
procedure shown below. Thin layer chromatography on silica gel was used to
separate the respiratory quinones into different classes, in which hexane:
tert¬butylmethylether (9:1 v/v) work as the solvent. After that, UV absorbing
bands which correspond to the different quinone classes were removed from
the plate and further analyzed by HPLC. This step is performed on a LDC
Analytical HPLC fitted with a reverse phase column using methanol: heptane
9:1 (v/v) as the mobile phase. Respiratory quinones are detected at 269 nm.