An integrated procedure for the extraction of bacterial isoprenoid quinones and polar lipids

Publisher Summary Recent advances in the biochemistry of microorganisms have revealed that cell-component analysis can be effectively applied to bacterial systematics, providing the basis of chemotaxonomy. Analyses of cell components are coming to be essential tools not only for bacterial classification but also for identification. The sequence of chromosomal DNA holds the essential genetic information for an organism. Phylogenetic relationships are often examined using data on nucleotide sequences of ribosomal RNA. However, it is still impossible to determine the sequences of large polynucleotides for the number of organisms sufficient for taxonomic purposes. DNA (RNA) homologies, which have been applied to various kinds of bacteria, present useful information on bacterial systematics. Homology indices are values used in comparing microorganisms. DNA homology data contribute to the clarification, not only of the relatedness of two organisms, but also of the evaluation of phenotypic characteristics.

4 Lipid and Cell-Wall Analysis in Bacterial Systematics

The species of the genus 'Thiobacillus' fall into the alpha-, beta- and gamma-subclasses of the Proteobacteria, the type species Thiobacillus thioparus being located in the beta-subclass. 'Thiobacillus' species exhibit almost as much diversity in DNA composition and physiology as is found collectively in all other proteobacterial groups. On the basis of physiological characters and 16S rRNA gene sequence comparisons, eight of the existing Thiobacillus species are proposed for reassignment to three newly designated genera within the gamma-subclass of the Proteobacteria, namely Acidithiobacillus, Halothiobacillus and Thermithiobacillus.

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Reclassification of some species of Thiobacillus to the newly designated genera Acidithiobacillus gen. nov., Halothiobacillus gen. nov. and Thermithiobacillus gen. nov.

Reverse-phase high-performance liquid chromatography and photodiode array detection were used to analyze the microbial quinones of influent sewage and activated sludge in a sewage treatment plant. Significant differences in quinone patterns were noted between the sewage and activated sludge. Unlike the activated sludge, the sewage had low ratios of menaquinones to ubiquinones, and contained menaquinone-6 as the most abundant menaquinone and negligible amounts of partially hydrogenated menaquinones. A photodiode array analysis revealed that the sewage also contained considerable amounts of plastoquinones and vitamin K,, both of which are specific to photosynthetic electron transport in cyanobacteria and chloroplasts. These results suggest that the microbial population structure of sewage is markedly different from that of activated sludge. Relationships between changes in the community structure of the sewage and activated sludge were also discussed on the basis of the results of a numerical analysis of lipoquinone patterns.

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Comparative lipoquinone analysis of influent sewage and activated sludge by high-performance liquid chromatography and photodiode array detection

Polyamine Pattern as a Chemotaxonomic Marker within the Proteobacteria

Menaquinone mixtures of microbial origin were separated according to the chain length and the degree of hydrogenation of the polyisoprenyl side-chain by reversephase partition high performance liquid chromatography. Menaquinones can be measured easily and sensitively by the chromatographic system described here.

Analysis of bacterial menaquinone mixtures by high performance liquid chromatography

In order to develop a new method for determination of the G+C content of a DNA, preparations from 26 bacterial strains and salmon sperm were digested to form 5'-deoxyribonucleotide-monophosphates (dNMP) with nuclease P1 and subjected to high performance liquid chromatography (HPLC). Chromatograms indicated excellent separation of the components, and the data analysis suggested sufficient reproducibility and reasonable A/T and G/C ratios over a wide range (39 to 72mol%) of G+C values.

Estimation of DNA base composition by high performance liquid chromatography of its nuclease P1 hydrolysate.

Summary: Facultatively chemolithotrophic thiobacilli. Thiobacillus perometabolis
thi 022, Thiobacillus sp. A2, Thiobacillus novellus, Thiobacillus organoparus and Thiobacillus acidophilus, had ubiquinones with 10 isoprene units (Q-10) (group I). These species also had [octadecenoic acid (18:1) + cyclopropane acid of C19 (19cyc)] as major non-hydroxylated fatty acids. The organisms, except T. novellus, contained 3-hydroxydecanoic acid (3-OH 10:0) or 3-hydroxytetradecanoic acid (3-OH 14:0) as hydroxy fatty acids. DNA base compositions of these species were 63 to 68 mol% G + C. Other facultatively chemolithotrophic thiobacilli, including T. perometabolis
thi 023, Thiobacillus delicatus and Thiobacillus intermedius contained Q-8 (group II). They had hexadecanoic acid (16:0), [hexadecenoic acid (16:1) + cyclopropane acid of C17 (17cyc)] and [18:1 + 19cyc] as major non-hydroxylated fatty acids. The species of group II had 3-OH 10:0 and some possessed 3-hydroxydodecanoic acid (3-OH 12:0). The DNA base composition of these strains was about 65 mol% G + C. Obligate chemolithotrophic thiobacilli contained Q-8 (group III). They had 16:0, [16:1 + 17cyc] with or without [18:1 + 19cyc] as major non-hydroxylated fatty acids. The species of group III contained 3-OH 12:0 and some had 3-OH 10:0 when grown at neutral pH and 3-OH 14:0 in acidophilic conditions. DNA base compositions of these organisms ranged from 51 to 64 mol% G + C. Chemotaxonomic characteristics would seem to be useful for the identification and the classification of thiobacilli.

Yoko Katayama-Fujimura

Papers

Analysis of bacterial menaquinone mixtures by high performance liquid chromatography

Estimation of DNA base composition by high performance liquid chromatography of its nuclease P1 hydrolysate.

Ubiquinone, Fatty acid and DNA Base Composition Determination as a Guide to the Taxonomy of the Genus Thiobacillus

Determination of the nucleotide composition of a deoxyribonucleic acid by high-performance liquid chromatography of its enzymatic hydrlysate: a review

Characteristics of H2S oxidizing bacteria inhabiting a peat biofilter