ubiquinone-9 and ubiquinone-8

A moderately halophilic, aerobic bacterium, strain BZ-SZ-XJ27T, belonging to the genus Halomonas, was isolated from a saline-alkaline lake in the Xinjiang Uyghur Autonomous Region of China. Phylogenetic analysis based on 16S rRNA gene sequences and a multilocus sequence analysis using the 16S rRNA, gyrB and rpoD genes demonstrated that strain BZ-SZ-XJ27T represents a member of the genus Halomonas. On the basis of 16S rRNA gene sequence similarity, the closest relatives were Halomonas campaniensis 5AGT, H. fontilapidosi 5CRT, H. korlensis XK1T and H. sinaiensis ALO SharmT, with similarities of 96.2-97.2 %. DNA-DNA hybridization with H. korlensis CGMCC 1.6981T (the nearest phylogenetic neighbour) and H. campaniensis DSM 15293T (the highest 16S rRNA gene sequence similarity) showed relatedness values of 53 and 38 %, respectively, demonstrating the separateness of the three taxa. The bacterium stained Gram-negative and the cells were motile and rod-shaped. The strain formed creamy-white colonies and grew under optimal conditions of 1.42 M Na+ (range 0.22-4.32 M Na+), pH 8.0-8.5 (range pH 6.0-10.0) and 39 °C (range 4-43 °C). The dominant fatty acids were summed feature 8 (C18 : 1ω7c/C18 : 1ω6c; 36.6 %), C16 : 0 (25.9 %) and summed feature 3 (C16 : 1ω7c/C16 : 1ω6c; 21.2 %). The dominant polar lipids were two unknown phospholipids, phosphatidylethanolamine and phosphatidylglycerol, and the main respiratory quinones were ubiquinone 9 (Q-9; 89 %) and ubiquinone 8 (Q-8; 10 %). The genomic DNA G+C content was 61.7 ± 0.8 mol% (Tm). On the basis of phenotypic, chemotaxonomic and phylogenetic features, strain BZ-SZ-XJ27T is proposed to represent a novel species, Halomonas urumqiensis sp. nov., within the genus Halomonas of the family Halomonadaceae. The type strain is BZ-SZ-XJ27T ( = JCM 30202T = CGMCC 1.12917T).

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Halomonadaceae; Halomonas; Hydrogen-Ion Concentration; Lakes; Multilocus Sequence Typing; Nucleic Acid Hybridization; Phospholipids; Phylogeny; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Ubiquinone; Water Microbiology

A Gram-staining-negative, rod-shaped, motile and facultatively anaerobic bacterial strain, designated X2(T), was isolated from the sludge of an anaerobic, denitrifying, sulfide-removal bioreactor, and found to oxidize sulfide anaerobically with nitrate as electron acceptor. The strain grew at salinities of 0-3% (w/v) NaCl (optimum, 0-1%). Growth occurred at pH 6.0-10.0 (optimum, pH 8.0) and 10-37 °C (optimum, 30 °C). The genomic DNA G+C content was 59 mol%. Q-8 and Q-9 were detected as the respiratory quinones. The major fatty acids (>10 %) were C16:1ω7c and/or C16: 1ω6c, C18: 1ω7c and C16:0. The polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and one unidentified phospholipid. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain X2(T) formed a novel clade within the family Pseudomonadaceae, with the highest sequence similarity to Pseudomonas caeni KCTC 22292(T) (93.5%). On the basis of phenotypic, chemotaxonomic and phylogenetic characteristics, it is proposed that this strain represents novel genus and species within the family Pseudomonadaceae, for which the name Thiopseudomonas denitrificans gen. nov., sp. nov. is proposed. The type strain is X2(T) ( =CCTCC M 2013362(T) =DSM 28679(T) = KCTC 42076(T)).

Topics: Bacterial Typing Techniques; Base Composition; Bioreactors; DNA, Bacterial; Fatty Acids; Molecular Sequence Data; Phospholipids; Phylogeny; Pseudomonadaceae; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Sewage; Ubiquinone

Two Gram-negative, facultatively anaerobic, rod-shaped bacteria, strains EF212(T) and PS125(T), were isolated from the octocorals Eunicea fusca and Plexaura sp., respectively. EF212(T) was isolated from a specimen of E. fusca collected off the coast of Florida, USA, and PS125(T) was isolated from a specimen of Plexaura sp. collected off the coast of Bimini, Bahamas. Analysis of the nearly full-length 16S rRNA gene sequences showed that these novel strains were most closely related to Endozoicomonas montiporae CL-33(T), E. elysicola MKT110(T) and E. numazuensis HC50(T) (EF212(T), 95.6-97.2 % identity; PS125(T), 95.1-96.4 % identity). DNA-DNA hybridization values among EF212(T), PS125(T), E. montiporae LMG 24815(T) and E. elysicola KCTC 12372(T) were far below the 70 % cut-off, with all values for duplicate measurements being less than 35 %. Both EF212(T) and PS125(T) required NaCl for growth and showed optimal growth at 2-3 % NaCl, 22-30 °C and pH 8.0. The predominant cellular fatty acids were summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c), summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c), C16 : 0 and C14 : 0. The DNA G+C content of EF212(T) was 48.6 mol% and that of PS125(T) was 47.5 mol%. In addition to the genotypic differences observed between the two novel strains and related type strains, phenotypic and chemotaxonomic experiments also revealed differences between strains. Thus, strains EF212(T) and PS125(T) represent novel species of the genus Endozoicomonas, for which the names Endozoicomonas euniceicola sp. nov. and Endozoicomonas gorgoniicola sp. nov., respectively, are proposed. The type strains are EF212(T) ( = NCCB 100458(T) = DSM 26535(T)) for Endozoicomonas euniceicola sp. nov. and PS125(T) ( = NCCB 100438(T) = CECT 8353(T)) for Endozoicomonas gorgoniicola sp. nov. An emended description of the genus Endozoicomonas is also provided to encompass differences observed in the results of genotypic, chemotaxonomic and phenotypic tests compared from the original and amended genus descriptions.

Topics: Animals; Anthozoa; Bacterial Typing Techniques; Bahamas; Base Composition; DNA, Bacterial; Fatty Acids; Florida; Gammaproteobacteria; Genotype; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

We studied ubiquinone (Q), Q homologue ratio, and steady-state levels of mCOQ transcripts in tissues from mice fed ad libitum or under calorie restriction. Maximum ubiquinone levels on a protein basis were found in kidney and heart, followed by liver, brain, and skeletal muscle. Liver and skeletal muscle showed the highest Q(9)/Q(10) ratios with significant interindividual variability. Heart, kidney, and particularly brain exhibited lower Q(9)/Q(10) ratios and interindividual variability. In skeletal muscle and heart, the most abundant mCOQ transcript was mCOQ7, followed by mCOQ8, mCOQ2, mPDSS2, mPDSS1, and mCOQ3. In nonmuscular tissues (liver, kidney, and brain) the most abundant mCOQ transcript was mCOQ2, followed by mCOQ7, mCOQ8, mPDSS1, mPDSS2, and mCOQ3. Calorie restriction increased both ubiquinone homologues and mPDSS2 mRNA in skeletal muscle, but mCOQ7 was decreased. In contrast, Q(9) and most mCOQ transcripts were decreased in heart. Calorie restriction also modified the Q(9)/Q(10) ratio, which was increased in kidney and decreased in heart without alterations in mPDSS1 or mPDSS2 transcripts. We demonstrate for the first time that unique patterns of mCOQ transcripts exist in muscular and nonmuscular tissues and that Q and COQ genes are targets of calorie restriction in a tissue-specific way.

Topics: Animals; Brain; Caloric Restriction; Free Radicals; Kidney; Liver; Mice; Muscle, Skeletal; Myocardium; Organ Specificity; RNA, Messenger; Ubiquinone

Coenzyme Q (Q) regulates aging in Caenorhabditis elegans, and its deficiency leads to a variety of pathologies in humans. We used a coq-8 deleted strain to study the role of Q in C. elegans development and how it influences life span. Endogenous Q(9) content of coq-8(ok840) knockouts was demonstrated to be about 7% of that found in the wild-type, indicating the basal biosynthesis rate is reduced in this strain. Knockouts abnormally developed both gonads and hypodermis, showed reduced fertility and shortened life span, and this was partially recovered by ingestion of exogenous Q. Knockouts produced embryos that showed arrested development at the time of initial expression of coq-8 in embryo. Uridine, whose biosynthesis depends on mitochondrial Q, improved both egg production and progeny under Q-rich dietary conditions. COQ-8 is a candidate protein for post-translational regulation of Q biosynthesis rate and its expression correlates with Q content during the life cycle in C. elegans. We show for the first time that a critical level of Q is necessary to support embryo development and fertility in C. elegans. These results suggest that extra-mitochondrial function of Q is a key factor linking development and bioenergetics in C. elegans.

Topics: Aging; Animals; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Fertility; Gene Expression Regulation, Developmental; Gene Expression Regulation, Enzymologic; Gene Knockout Techniques; Genotype; Gonads; Larva; Longevity; Phenotype; Ubiquinone; Uridine

A procedure was developed to isolate fractions enriched in plasma membrane from Caenorhabditis elegans. Coenzyme Q9 (Q9) was found in plasma membrane isolated from either wild-type or long-lived qm30 and qm51 clk-1 mutant strains of Caenorhabditis elegans, along with dietary coenzyme Q8 (Q8) and the biosynthetic intermediate demethoxy-Q9 (DMQ9). NADH was able to reduce both Q8 and Q9, but not DMQ9. Our results indicate that DMQ9 cannot achieve the same redox role of Q9 in plasma membrane, suggesting that proportion of all these Q isoforms in plasma membrane must be an important factor in establishing the clk-1 mutant phenotype.

Topics: Animals; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Cell Membrane; NAD; Oxidation-Reduction; Ubiquinone

Ubiquinone (coenzyme Q; Q) is a key factor in the mitochondria electron transport chain, but it also functions as an antioxidant and as a cofactor of mitochondrial uncoupling proteins. Furthermore, Q isoforms balance in Caenorhabditis elegans is determined by both dietary intake and endogenous biosynthesis. In the absence of synthesis, withdrawal of dietary Q8 in adulthood extends life span. Thus, Q plays an important role in the aging process and understanding its synthesis acquires a new impetus. We have identified by RNA interference (RNAi) eight genes, including clk-1, involved in ubiquinone biosynthesis in C. elegans feeding animals with dsRNA-containing Escherichia coli HT115 strains. Silenced C. elegans showed lower levels of both endogenous Q9 and Q8 provided by diet, produced less superoxide without a significant modification of mitochondrial electron chain, and extended life span compared with non-interfered animals. E. coli strains harboring dsRNA also interfered with their own Q8 biosynthesis. These findings suggest that more efficient electron transport between a lower amount of Q and electron transport capacity of the mitochondrial complexes leads to less production of reactive oxygen species that contributes to extension of life span in the nematode C. elegans.

Topics: Animals; Caenorhabditis elegans; Electron Transport; Escherichia coli; Longevity; Mitochondria; Models, Biological; RNA Interference; Superoxides; Transformation, Bacterial; Ubiquinone

Topics: Aging; Animals; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Cell Nucleus; Diet; Electron Transport; Energy Metabolism; Escherichia coli; Fermentation; Helminth Proteins; Insulin; Larva; Longevity; Mitochondria; Mutation; Reactive Oxygen Species; Receptor, Insulin; Receptors, Cytoplasmic and Nuclear; Signal Transduction; Ubiquinone

The isoprenylated benzoquinone coenzyme Q is a redox-active lipid essential for electron transport in aerobic respiration. Here, we show that withdrawal of coenzyme Q (Q) from the diet of wild-type nematodes extends adult life-span by approximately 60%. The longevity of clk-1, daf-2, daf-12, and daf-16 mutants is also extended by a Q-less diet. These results establish the importance of Q in life-span determination. The findings suggest that Q and the daf-2 pathway intersect at the mitochondria and imply that a concerted production coupled with enhanced scavenging of reactive oxygen species contributes to the substantial life-span extension.

Topics: Aging; Animals; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Diet; Escherichia coli; Fermentation; Forkhead Transcription Factors; Genes, Helminth; Helminth Proteins; Larva; Longevity; Mitochondria; Models, Biological; Mutation; Oxidation-Reduction; Oxygen Consumption; Phenotype; Reactive Oxygen Species; Receptor, Insulin; Receptors, Cytoplasmic and Nuclear; Signal Transduction; Transcription Factors; Ubiquinone

The thermotropic properties of coenzymes Q10, Q9, Q8, and Q7 have been examined by differential scanning calorimetry and wide-angle X-ray diffraction. Typical scanning calorimetry cooling curves of coenzyme Q from the liquid state exhibit a single exothermic phase transition into a crystalline state at a temperature that decreases as the length of the polyisoprenoid side-chain substituent decreases. Upon subsequent heating, the molecules undergo a series of thermal events which precede the main crystalline-to-liquid endothermic phase transition. The temperature of these transitions increases with increasing chain length. The crystallization phase transition temperature depends markedly on the rate at which the sample is cooled and increases with decreasing scan rate; the temperature of the melting endotherm is not markedly affected by the scan rate. Detailed calorimetric studies of coenzyme Q10 indicate that two crystalline states are formed, one at relatively high cooling rates to low temperatures and the other when preparations are cooled slowly from the liquid state to relatively high temperatures. Heating the crystalline phase formed by rapid cooling causes its transformation into the phase observed by cooling slowly. X-ray diffraction analysis confirmed the existence of these two crystal phases in coenzymes Q9 and Q10 and the transformation from the rapidly crystallized form to the more ordered form associated with slower cooling rates. At body temperature (310 K) under equilibrium conditions coenzyme Q10 exists in an ordered crystalline phase; the implications of the thermotropic behavior of coenzyme Q10 on mitochondrial function in vitro and in vivo are discussed.

Topics: Calorimetry, Differential Scanning; Coenzymes; Crystallization; Crystallography, X-Ray; Mitochondria; Thermodynamics; Ubiquinone

Topics: Animals; Anti-Infective Agents; Coenzymes; Female; Immunity, Cellular; Listeriosis; Mice; Spleen; Ubiquinone