Methylobacterium soli sp. nov. a methanol-utilizing bacterium isolated from the forest soil

A Gram-negative, pink-pigmented, non-spore-forming rod shaped, methanol-utilizing bacterium, strain YIM 48816(T), was isolated from forest soil collected from Sichuan province, China. Strain YIM 48816(T) can grow at 4-37 °C, pH 5.0-7.0 and 0% NaCl (w/v). Based on 16S rRNA gene sequence similarity studies, it belonged to the genus Methylobacterium, and formed a phyletic line. The 16S rRNA gene sequence similarities were 96.2% to Methylobacterium mesophilicum DSM 1708(T) and 96.0% to Methylobacterium brachiatum DSM 19569(T), and the phylogenetic similarities to all other Methylobacterium species with validly published names were less than 96.0%. The major menaquinones detected were Q-10 (97.14%) and Q-9 (2.86%). The major fatty acids were C18:1 ω7c (80.84%). The DNA G + C content was 66.2 mol%. It is apparent from the genotypic and phenotypic data that strain YIM 48816(T) belongs to a novel species of the genus Methylobacterium, for which the name Methylobacterium soli sp. nov. is proposed. The type strain is YIM 48816(T) (CCTCC AA 208027(T) = KCTC 22810(T)).     

Methylopila helvetica sp. nov. and Methylobacterium dichloromethanicum sp. nov.--novel aerobic facultatively methylotrophic bacteria utilizing dichloromethane

Eight strains of Gram-negative, aerobic, asporogenous, neutrophilic, mesophilic, facultatively methylotrophic bacteria are taxonomically described. These icl- serine pathway methylobacteria utilize dichloromethane, methanol and methylamine as well as a variety of polycarbon compounds as the carbon and energy source. The major cellular fatty acids of the non-pigmented strains DM1, DM3, and DM5 to DM9 are C18:1, C16:0, C18:0, Ccy19:0 and that of the pink-pigmented strain DM4 is C18:1. The main quinone of all the strains is Q-10. The non-pigmented strains have similar phenotypic properties and a high level of DNA-DNA relatedness (81-98%) as determined by hybridization. All strains belong to the alpha-subgroup of the alpha-Proteobacteria. 16S rDNA sequence analysis led to the classification of these dichloromethane-utilizers in the genus Methylopila as a new species - Methylopila helvetica sp.nov. with the type strain DM9 (=VKM B-2189). The pink-pigmented strain DM4 belongs to the genus Methylobacterium but differs from the known members of this genus by some phenotypic properties, DNA-DNA relatedness (14-57%) and 16S rDNA sequence. Strain DM4 is named Methylobacterium dichloromethanicum sp. nov. (VKM B-2191 = DSMZ 6343).     

Paracoccus aminophilus sp. nov. and Paracoccus aminovorans sp. nov., which utilize N,N-dimethylformamide

Two methylamine- and N,N-dimethylformamide-utilizing Paracoccus spp. are described. These bacteria are gram-negative, nonsporeforming, nonmotile, coccoid or short rod-shaped organisms. Their DNA base composition is 62 to 68 mol% G + C. Their cellular fatty acids include large amounts of C18:1 acid. Their major hydroxy acids are 3-OH C10:0 and 3-OH C14:0 acids. The major ubiquinone is Q-10. These bacteria are distinguished from Paracoccus denitrificans and Paracoccus alcaliphilus by physiological characteristics and by DNA-DNA-homology. Paracoccus aminophilus sp. nov. and Paracoccus aminovorans sp. nov. are proposed. The type strain of P. aminophilus is DM-15 (= JCM 7686), and the type strain of P. aminovorans is DM-82 (= JCM 7685). Paracoccus halodenitrificans is distinguished from other Paracoccus species on the basis of cellular fatty acid composition, hydroxy fatty acid composition, and DNA-DNA homology. It may not be a valid member of the genus Paracoccus.     

<Emphasis Type="Italic">Methylobacterium dankookense</Emphasis> sp. nov., isolated from drinking water

A pink-pigmented bacterium, designated SW08-7^T was isolated from the drinking water of a water purifier. Cells were Gram-negative, rod-shaped, strictly aerobic, and non-spore-forming. It grew optimally at 25°C, pH 6∼7. Phylogenese analysis based on 16S rRNA gene sequence showed that strain SW08-7^T belongs to the genus Methylobacterium. The highest 16S rRNA gene sequence similarities were found to Methylobacterium mesophilicum JCM 2829^T (96.9%), Methylobacterium brachiatum B0021^T (96.9%), Methylobacterium phyllosphaerae CBMB27^T (96.6%), Methylobacterium radiotolerans JCM 2831^T (96.6%), and Methylobacterium hispanicum GP34^T (96.5%). DNA-DNA hybridization experiment revealed low-level (28.5%) of DNA-DNA relatedness between strain SW08-7^T and Methylobacterium hispanicum. The genomic DNA G+C content was 68.9 mol% and the major isoprenoid quinone was Q-10. The major cellular fatty acid of strain SW08-7^T was C_18:1 ω7c (79.8±2.1%). Results of phylogenetic, phenotypic, and biochemical analyses revealed that strain SW08-7^T could be classified as representing a novel species of genus Methylobacterium, for which the name Methylobacterium dankookense sp. nov. is proposed. The type strain is SW08-7^T (=KCTC 22512^T =DSM 22415¹).     

Reclassification of Methylobacterium chloromethanicum and Methylobacterium dichloromethanicum as later subjective synonyms of Methylobacterium extorquens and of Methylobacterium lusitanum as a later subjective synonym of Methylobacterium rhodesianum

Phylogenetic analysis based on 16S rDNA sequences was performed on all type strains of the 14 validly described Methylobacterium species to ascertain the genealogic relationships among these species. The results showed that type strains of Methylobacterium were divided into two monophyletic groups whose members were distinct species with sequence similarity values greater than 97.0% between any two of the members in the same group. Only M. organophilum JCM 2833(T) and ATCC 27886(T) were not divided into those two groups. In particular, strains of M. dichloromethanicum and M. chloromethanicum exhibited extremely high similarity values (99.9 and 100%, respectively) with the type strain of M. extorquens. To clarify the relationships among Methylobacterium species in more detail, phylogenetic analysis based on the 5' end hyper-variable region of 16S rDNA (HV region), ribotyping analysis, fatty acid analysis, G+C content analysis and DNA-DNA hybridization experiments was performed on 58 strains of Methylobacterium species. Results of the ribotyping analysis and the phylogenetic analysis based on HV region sequences indicated that many Methylobacterium strains, including M. 'organophilum' DSM 760(T), have been erroneously identified. The DNA G+C content of Methylobacterium strains were between 68.1 and 71.3%. Results of whole-cell fatty-acid profiles showed that all strains contained 18 : 1omega7c as the primary fatty acid component (82.8-90.1%), with 16 : 0 and 18 : 0 as minor components. M. dichloromethanicum DSM 6343(T), M. chloromethanicum NCIMB 13688(T), and M. extorquens IAM 12631(T) exhibited high DNA-DNA relatedness values between each other (69-80%). M. lusitanum NCIMB 13779(T) also showed a close relationship with M. rhodesianum DSM 5687(T) at DNA-DNA relatedness levels of 89-92%. According to these results, many Methylobacterium strains should be reclassified, with M. dichloromethanicum and M. chloromethanicum regarded as a synonym of M. extorquens, and M. lusitanum a synonym for M. rhodesianum.     

Phospholipid Metabolism in Ferrobacillus ferrooxidans

The lipid composition of the chemoautotroph Ferrobacillus ferrooxidans has been examined. Fatty acids represent 2% of the dry weight of the cells and 86% of the total are extractable with organic solvents. About 25% of the total fatty acids are associated with diacyl phospholipids. Polar carotenoids, the benzoquinone coenzyme Q-8, and most of the fatty acids are present in the neutral lipids. The phospholipids have been identified as phosphatidyl monomethylethanolamine (42%), phosphatidyl glycerol (23%), phosphatidyl ethanolamine (20%), cardiolipin (13%), phosphatidyl choline (1.5%), and phosphatidyl dimethylethanolamine (1%) by chromatography of the diacyl lipids, by chromatography in four systems of the glycerol phosphate esters derived from the lipids by mild alkaline methanolysis, and by chromatographic identification of the products of acid hydrolysis of the esters. No trace of phosphatidylserine (PS), glycerolphosphorylserine, or serine could be detected in the lipid extract or in derivatives of that extract. This casts some doubt on the postulated involvement of PS in iron metabolism. After growth in the presence of (14)C and (32)P, there was essentially no difference in the turnover of either isotope in the glycerolphosphate ester derived from each lipid in cells grown at pH 1.5 or 3.5.     

The respiratory chain in a ubiquinone-deficient mutant of Saccharomyces cerevisiae.

1. Two allelic mutants of Saccharomyces cerevisiae with a deficiency in the biosynthesis of ubiquinone have been isolated. The properties of one particular mutant strain were investigated. Submitochondrial particles of this strain contain maximally 3% of the amount of ubiquinone in wild-type particles; the amounts of other components of the respiratory chain are essentially normal. 2. The respiratory rates of mutant cells, mitochondria and submitochondrial particles are low with ubiquinone-dependent substrates, but are restored to normal levels by addition of Q-1; the restored respiration is antimycin sensitive. Intact cells and mitochondria show respiratory control both in the absence and presence of Q-1. 3. The NADH:Q-1 oxidoreductase of submitochondrial particles of the mutant followspseudo first-order kinetics in [Q-1]. QH2-1 inhibits competitively with respect to Q-1, the Ki for QH2-1 being equal to the Km for Q-1. 4. Succinate dehydrogenase in both wild-type and mutant submitochondrial particles can be activated by NADH. 5. The turnover number of succinate dehydrogenase in the mutant, measured with phenazine methosulphate as primary electron acceptor, is about one-half that of wild-type particles. The turnover numbers measured with Q-1 as electron acceptor are about the same in the two types of particles. 6. The kinetics of redox changes in cytochrome b, in the presence of antimycin and oxygen, are distinctly different in the mutant and wild-type particles. They indicate that ubiquinone plays an important role in the phenomenon of the increased reducibility of cytochrome b induced by antimycin plus oxygen.     

16S ribosomal RNA sequence analysis for determination of phylogenetic relationship among methylotrophs.

16S ribosomal RNAs (rRNA) of 12 methylotrophic bacteria have been almost completely sequenced to establish their phylogenetic relationships. Methylotrophs that are physiologically related are phylogenetically diverse and are scattered among the purple eubacteria (class Proteobacteria). Group I methylotrophs can be classified in the beta- and the gamma-subdivisions and group II methylotrophs in the alpha-subdivision of the purple eubacteria, respectively. Pink-pigmented facultative and non-pigmented obligate group II methylotrophs form two distinctly separate branches within the alpha-subdivision. The secondary structures of the 16S rRNA sequences of 'Methylocystis parvus' strain OBBP, 'Methylosinus trichosporium' strain OB3b, 'Methylosporovibrio methanica' strain 81Z and Hyphomicrobium sp. strain DM2 are similar, and these non-pigmented obligate group II methylotrophs form one tight cluster in the alpha-subdivision. The pink-pigmented facultative methylotrophs, Methylobacterium extorquens strain AM1, Methylobacterium sp. strain DM4 and Methylobacterium organophilum strain XX form another cluster within the alpha-subdivision. Although similar in phenotypic characteristics, Methylobacterium organophilum strain XX and Methylobacterium extorquens strain AM1 are clearly distinguishable by their 16S rRNA sequences. The group I methylotrophs, Methylophilus methylotrophus strain AS1 and methylotrophic species DM11, which do not utilize methane, are similar in 16S rRNA sequence to bacteria in the beta-subdivision. The methane-utilizing, obligate group I methanotrophs, Methylococcus capsulatus strain BATH and Methylomonas methanica, are placed in the gamma-subdivision. The results demonstrate that it is possible to distinguish and classify the methylotrophic bacteria using 16S rRNA sequence analysis.     

A comparison of the major lipid classes and fatty acid composition of marine unicellular cyanobacteria with freshwater species

The fatty acid composition of two motile (strains WH 8113 and WH 8103) and one nonmotile (strain WH 7803) marine cyanobacteria has been determined and compared with two freshwater unicellular Synechocystis species (strain PCC 6308 and PCC 6803). The fatty acid composition of lipid extracts of isolated membranes from Synechocystis PCC 6803 was found to be identical to that of whole cells. All the marine strains contained myristic acid (14:0) as the major fatty acid, with only traces of polyunsaturated fatty acids. This composition is similar to Synechocystis PCC 6308. The major lipid classes of the nonmotile marine strain were identified as digalactosyl diacylglycerol, monogalactosyl diacylglycerol, phosphatidylglycerol, and sulfoquinovosyl diacylglycerol, identical to those found in other cyanobacteria.Abbreviations DGDG Digalactosyl diacylglycerol - MGDG Monogalactosyldiacylglycerol - PG Phosphatidylglycerol - SGDG sulfoquinovosyl diacylglycerol - gc gas chromatography - ms mass spectrometry     

Biodegradation of nitro-substituted explosives 2,4,6-trinitrotoluene, hexahydro-1,3,5-trinitro-1,3,5-triazine, and octahydro-1,3,5,7-tetranitro-1,3,5-tetrazocine by a phytosymbiotic Methylobacterium sp. associated with poplar tissues (Populus deltoides x nigra DN34)

A pink-pigmented symbiotic bacterium was isolated from hybrid poplar tissues (Populus deltoides x nigra DN34). The bacterium was identified by 16S and 16S-23S intergenic spacer ribosomal DNA analysis as a Methylobacterium sp. (strain BJ001). The isolated bacterium was able to use methanol as the sole source of carbon and energy, which is a specific attribute of the genus Methylobacterium. The bacterium in pure culture was shown to degrade the toxic explosives 2,4,6-trinitrotoluene (TNT), hexahydro-1,3,5-trinitro-1,3,5-triazene (RDX), and octahydro-1,3,5,7-tetranitro-1,3,5-tetrazocine (HMX). [U-ring-(14)C]TNT (25 mg liter(-1)) was fully transformed in less than 10 days. Metabolites included the reduction derivatives amino-dinitrotoluenes and diamino-nitrotoluenes. No significant release of (14)CO(2) was recorded from [(14)C]TNT. In addition, the isolated methylotroph was shown to transform [U-(14)C]RDX (20 mg liter(-1)) and [U-(14)C]HMX (2.5 mg liter(-1)) in less than 40 days. After 55 days of incubation, 58.0% of initial [(14)C]RDX and 61.4% of initial [(14)C]HMX were mineralized into (14)CO(2). The radioactivity remaining in solution accounted for 12.8 and 12.7% of initial [(14)C]RDX and [(14)C]HMX, respectively. Metabolites detected from RDX transformation included a mononitroso RDX derivative and a polar compound tentatively identified as methylenedinitramine. Since members of the genus Methylobacterium are distributed in a wide diversity of natural environments and are very often associated with plants, Methylobacterium sp. strain BJ001 may be involved in natural attenuation or in situ biodegradation (including phytoremediation) of explosive-contaminated sites.     

Isolation and nucleotide sequence of the methanol dehydrogenase structural gene from Paracoccus denitrificans.

A genomic clone bank of Paracoccus denitrificans DNA has been constructed in the expression vector set pEX1, pEX2, and pEX3. Screening of this clone bank with antibodies raised against P. denitrificans methanol dehydrogenase resulted in the isolation of a clone, pNH3, that synthesized methanol dehydrogenase cross-reactive proteins. The nucleotide sequence of the P. denitrificans DNA fragment inserted in this clone has been determined and shown to contain the full methanol dehydrogenase structural gene. DNA cross-hybridization was found with DNA fragments which have been reported to contain the methanol dehydrogenase structural genes from Methylobacterium sp. strain AM1 and Methylobacterium organophilum.     

On the role of Fe2+ in bacterial photosynthesis. The effect of biosynthetic substitution of Fe2+ by Mn2+ on the electron transfer step Q-1Q2----Q1Q-2 in reaction centers

A test of the 'iron-wire' hypothesis for the role of Fe2+ in promoting the electron transfer between the primary (Q1) and secondary (Q2) quinones in bacterial reaction centers of Rhodopseudomonas sphaeroides strain R-26.1 has been conducted. Kinetics of this step, P+Q-1Q2----P+Q1Q-2, and of recombination with the oxidized donor, P+Q-1----PQ1 and P+Q-2----PQ2, were followed optically at 4 degrees C in normal iron-containing reaction centers and in reaction centers having 58% Mn2+, replacing Fe2+. This significant replacement is accomplished biosynthetically by control of the growth conditions, and so should preserve the native interactions between the cofactors. There are no significant differences observed in the recombination kinetics of the two types of reaction centers. The electron transfer between the quinones was observed to show apparent biphasic kinetics with major components of approx. 170 microseconds and 1.5 ms at 4 degrees C and pH = 7.5. There is no statistically significant difference observed between the two types of reaction centers. This major change in the electronic structure of the metal and the unaltered kinetics discount the likelihood of any direct orbital participation of the metal in the electron transfer between the quinones.     

Genetic relatedness among species in Aspergillus section Clavati as measured by electrophoretic comparison of enzymes,DNA base composition,and DNA-DNA hybridization

Aspergillus subgenus Clavati has four recognized species: A. clavatus (the type species), A. clavatonanicus, A. giganteus, and A. longivesica. These species are strictly anamorphic (mitotic) and defined by the morphological species concept. However, their genealogical relationships remain uncertain. In this study, we examined the genetic relatedness among the four species in this section, using electrophoretic comparison of enzymes, DNA base composition, and DNA-DNA hybridization. In a dendrogram based on the calculated similarity values of four enzymes, 10 strains in section Clavati, 3 strains in the xerophilic species, a strain in section Ornati, and a strain in section Cremei were separated into nine major clusters at a 60% similarity level. A. longivesica JCM 10186(T) had Q-10 in our analysis, but Kuraishi et al. (1990) reported A. longivesica JCM 1720(T) had Q-9 (49%) and Q-10 (46%). The G+C contents of the four species of section Clavati ranged from 48 to 50 mol%. The degree of the intraspecific reassociation among the DNAs from the strains of these species ranged from 77 to 99%, whereas the degrees of interspecific relative binding among strains of the four species ranged from 30 to 59%. Our data from enzyme patterns and DNA relatedness support the validity of the three species in section Clavati, except for A. longivesica.     

Alterations in the phospholipid composition of Rhodopseudomonas sphaeroides and other bacteria induced by Tris.

Alterations in the phospholipid head group composition of most strains of Rhodopseudomonas sphaeroides, as well as Rhodopseudomonas capsulata and Paracoccus denitrificans, occurred when cells were grown in medium supplemented with Tris. Growth of R. sphaeroides M29-5 in Tris-supplemented medium resulted in the accumulation of N-acylphosphatidylserine (NAPS) to as much as 40% of the total whole-cell phospholipid, whereas NAPS represented approximately 28 an 33% of the total phospholipid when R. capsulata and P. denitrificans respectively, were grown in medium containing 20 mM Tris. The accumulation of NAPS occurred primarily at the expense of phosphatidylethanolamine in both whole cells and isolated membranes of R. sphaeroides and had no detectable effect on cell growth under either chemoheterotrophic or photoheterotrophic conditions. Yeast extract (0.1%) and Casamino Acids (1.0%) were found to be antagonistic to the Tris-induced (20 mM) alteration in the phospholipid composition of R. sphaeroides. The wild-type strains R. sphaeroides 2.4.1 and RS2 showed no alteration in their phospholipid composition when they were grown in medium supplemented with Tris. In all strains of Rhodospirillaceae tested, as well as in P. denitrificans, NAPS represented between 1.0 and 2.0% of the total phospholipid when cells were grown in the absence of Tris. [32P]orthophosphoric acid entered NAPS rapidly in strains of R. sphaeroides that do (strain M29-5) and do not (strain 2.4.1) accumulate this phospholipid in response to Tris. Our data indicate that the phospholipid head group composition of many Rhodospirillaceae strains, as well as P. denitrificans, is easily manipulated; thus, these bacteria may provide good model systems for studying the effects of these modifications on membrane structure and function in a relatively unperturbed physiological system.     

Pectinatus portalensis nov. sp., a relatively fast-growing, coccoidal, novel Pectinatus species isolated from a wastewater treatment plant

The genus Pectinatus is currently composed by two species, Pectinatus cerevisiiphilus and Pectinatus frisingensis , both asociated with beer spoilage. This study describes a novel isolate (strain B6) retrieved from a wastewater treatment plant collecting residues from a large number of wineries. Based on similarity analysis of 16S rRNA gene sequences, strain B6 belongs to the genus Pectinatus . Strain B6 is a strict anaerobe like other Pectinatus species and it presents non-motile, coccoid cells showing a slight oval shape. Strain B6 shows marked physiological differences with other Pectinatus species both in fatty acid composition and carbon source utilization. The most abundant fatty acids found in strain B6 were 18:1 (42.8%) and 16:0 (18.3%) representing a total of over 61% of fatty acids in this microorganism while these fatty acids represented 41.3% in P. cerevisiiphilusT and 2.4% in P. frisingensisT of their total. Fatty acid 15:0 was not significant in strain B6 and represented 28.6% and 13.3% for P. cerevisiiphilusT and P. frisingensisT, respectively. Strain B6 showed a faster growth rate and higher optimum temperature than its relatives P. cerevisiiphilus and P. frisingensis . Strain B6, P. cerevisiiphilus and P. frisingensis could be clearly differentiated by acid production tests from substrates such as esculine and gluconate, and the lack of acid production from rhamnose and fucose among others. G+C mol% content in strain B6 is 36.5%. Based on genotypic and phenotypic differences, strain B6 is proposed as a novel Pectinatus species, P. portalensis nov. sp. Both strain B6 and the two described species of Pectinatus grow on beers and wines. These results provide insights about the origin and reservoirs of Pectinatus species and spoiling alcoholic beverages.     

Transposon-mediated integration of the RP1 plasmid into chromosomes of methylotrophic bacteria

The homology region between the DNA of plasmid RP1ts::Tn601 and chromosome of the thermotolerant methylotrophic bacterium Methylobacterium sp. SKF240 has been constructed by transposon Tn601 translocation into the chromosome. The clones of Methylobacterium sp. SKF240 having integrated the plasmid RP1 into the chromosome have been obtained by conjugation on the basis of above mentioned genetic technique. The integration of plasmid RP1 into the chromosomal DNA of the methylotroph has been confirmed by the genetic and electrophoretic methods. Clones harbouring the integrated plasmid are able to transfer the chromosomal genes for methionine and isoleucine-valine synthesis to the recipient cells of P. aeruginosa PAO ML4262 by conjugation.     

Plasmid analysis in pink facultative methylotrophic bacteria using a modified acetone-alkaline hydrolysis method

Routine screening of indigenous and recombinant plasmids in pink facultative methylotrophic bacteria has been difficult, time-consuming, and yields variable results. We report a modified alkaline hydrolysis method for rapid plasmid isolation from these organisms that reproducibly results in good yields of closed circular plasmid DNA which can be readily digested with restriction enzymes. This method greatly facilitates direct screening of indigenous and introduced recombinant plasmids in the methylotrophic host strain. We have confirmed earlier findings that the original NCIB wild-type strain of Methylobacterium sp. strain AM1 (NCIB 9133) contains three cryptic plasmids. However, sizing of these plasmids by comparison to standards and by restriction fragment analysis suggests that they are larger than previously reported. We have designated these plasmids pAM1-1 (65 kb), pAM1-2 (40 kb) and pAM1-3 (33 kb). We have also shown that a rifamycin-resistant strain of Methylobacterium sp. strain AM1 used routinely in our laboratory lacks pAM1-2, although no phenotype has been associated with its loss. Finally, we have shown that another pink facultative methylotroph, Methylobacterium isolate (#YK1), contains three cryptic plasmids of approximately 43, 37 and 22 kb, respectively.     

Cytogenetic Analysis of Bone Marrow and Peripheral Blood Samples Stored in Fixative for Several Years

We have developed a method which improves the spreading of chromosomes and permits banding analysis of cytogenetic samples of bone marrow and unstimulated peripheral blood which have been stored in fixative for up to 15 years. Metaphase cells had been harvested as usual and stored in fixative (acetic acid:methanol 1:3) at -15 C. The procedure includes 4-5 changes of fixative (acetic acid:ethanol 1:1). Next, cells are dropped onto a chilled, wet slide. The back of the slide is then rinsed with 70% ethanol and dried by ignition. C-, G-, Q-, or R-banding patterns can now be obtained with these specimens. The procedure is useful for reinvestigation of cytogenetic samples that were obtained prior to the development of banding techniques.     

Production of isoprenoid compounds in the facultative methylotroph Protomonas extorquens

Screening of a strain which contained a large amount of ubiquinone Q-10 and a variety of isoprenoid compounds using different culture conditions and mutations was carried out.Protomonas extorquens TK 0045, which was found to contain carotenoid pigments, Hop-22(29)-ene, and Hopan-22-ol, was selected on the basis of cell yield and the content of ubiquinone Q-10. The contents of ubiquinone and sterols increased as the age of the culture increased, and reached a maximum level during the stationary phase.The contents of ubiquinone, sterols and carotenoid pigments, and ubiquinone homologs produced by P. extorquens TK 0045 were varied using mutagenesis. Mutants that had increased or decreased contents of carotenoid pigments were obtained with a high frequency. Most mutants had varying contents of other isoprenoid compounds. The ubiquinone homologs obtained by mutagenesis varied with a high frequency, and mutants which possessed increased levels of ubiquinone Q-9, Q-11 or Q-12 were isolated. However, the major ubiquinone component in these mutants was Q-10 the same as that in the wild strain. The production of ubiquinone was increased considerablyby repeated mutagenesis, with the content of ubiquinone produced by the third generation mutant (strains HB-5) being approximately 3.3 mg·g dry cell⁻¹ (2.5 times that of the wild strain). The acquisition of mutants exhibiting altered synthesis of carotenoid pigments would be useful for increasing the content of ubiquinone Q-10 in bacterial cells.