Growth and H2 production by Synechococcus spp. using organic/inorganic electron donors

Growth with simultaneous photoproduction of H₂ was obtained on various organic and inorganic compounds using axenic cultures of the oxygenic phototrophic bacteria Synechococcus sp. OU 103 and S. cedrorum. Highest H₂ production occurred with resting cells of S. cedrorum on malate (11.8 mmol H₂/vessel), whereas Synechococcus sp. OU 103 preferred sulphide (10.3 mmol H₂/vessel) as electron donor.The authors are with the Microbial Biotechnology Lab. Department of botany, Osmania University, Hyderabad 500 007, India.     

Alga-lytic activity of Pseudomonas fluorescens against the red tide causing marine alga Heterosigma akashiwo (Raphidophyceae)

A bacterial strain, HAK-13, exhibited strongest activity against Heterosigma akashiwo and was capable of controlling this bloom forming phytoplankton. Based on 16S rDNA sequences and biochemical and morphological characteristics, the strain HAK-13 was determined to be Pseudomonas fluorescens on the basis of 99.9% similarity with reference strains in the DNA databases. The growth of H. akashiwo was strongly suppressed by HAK-13 in all growth phases, with the strongest alga-lytic activity noted against harmful bloom-forming species in the exponential stage (6–22 days). Host range tests showed that HAK-13 also significantly inhibited the growth of Alexandrium tamarense and Cochlodinium polykrikoides but could not destroy Gymnodinium catenatum. P. fluorescens HAK-13 indirectly attacked H. akashiwo by alga-lytic substances that might be located at the compartment of cytoplasmic membrane of the bacterium at a level of 45.86 units/mg of specific activity. The results indicated that P. fluorescens HAK-13 caused cell lysis and death of H. akashiwo, A. tamarense, and C. polykrikoides dramatically and Prorocentrum dentatum slightly. Therefore, P. fluorescens HAK-13 has potential for use as a selective biocontrol of harmful algal blooms.     

Increased H2 production by immobilized microorganisms

Viable cells of H₂-producers (Bacillus licheniformis and a mixed microbial culture) were immobilized on brick dust and in calcium alginate beads. In batch culture, cells of the mixed culture in the free state yielded 8.2 l H₂/mol glucose utilized, whereasB. licheniformis evolved 13.1 l H₂. Immobilized cells, however, gave 4-fold more H₂ than the free bacteria. Highest yields were from the cells immobilized on brick dust. High H₂-production rates continued over two rounds of re-use of the immobilized cells.A. Kumar, S.R. Jain and A.P. Joshi were and V. C. Kalia is with the Centre for Biochemical Technology (CSIR), University Campus, Mall Road, Delhi-110 007, India; C.B. Sharma is with the Department of Biosciences and Biotechnology, University of Roorkee, Roorkee-247 667, India. A. Kumar is now with the Biochemistry Laboratory, Department of Chemistry, Indian Institute of Technology, New Delhi-110 016, India, S.R. Jain is now with the Centre for Biomedical Engineering, Indian Institute of Technology, New Delhi-110 016, India, and A.P. Joshi is now with the Chemical Engineering Division, National Chemical Laboratory, Pune-411 008, India.     

Spectroscopic and potentiometric characterization of cytochromes in two Sporomusa species and their expression during growth on selected substrates

The homoacetogenic bacteria Sporomusa ovata and Sporomusa sphaeroides were grown on betaine, betaine + formate, and acetoin in the absence of carbon dioxide, and the formation of membrane-bound cytochromes was determined. In S. sphaeroides, the growth substrate had little influence on the expression of cytochromes. In contrast, membranes from betaine-or acetoin-grown S. ovata cells had an 11-or 3-fold higher cytochrome b content than cells grown on betaine + formate. The cytochrome c content was reduced below the detection level after growth on the latter two substrates. The cytochromes in the membranes of S. sphaeroides and S. ovata were characterized by low-temperature difference spectroscopy, hemochrome difference spectroscopy, and redox potentiometry. Membranes of S. ovata were shown to contain two b-type cytochromes with E_m,7=-153±10 mV and E_m,7=-226±14 mV and two c-type cytochromes with E_m,7=-86±6 mV and E_m,7=-265±10 mV. In S. sphaeroides also two b-type cytochromes with E_m,7=-165±7 mV and E_m,7=-241±2 mV and two c-type cytochromes with E_m,7=-101±4 mV and E_{m, 8.5}=-338±9 mV could be distinguished. Cell extracts of S. sphaeroides were shown to contain all the enzymes of the acetyl-CoA (Wood) pathway. The degradation pathways of the substrates tested and the possible role of the cytochromes are discussed.Abbreviations E_m,7 midpoint potential at pH 7 and 25°C - H₄F tetrahydrofolate     

The unique root-nodule symbiosis between Rhizobium and the aquatic legume,Neptunia natans (L. f.) Druce

We examined the development of the aquatic N₂-fixing symbiosis between Rhizobium sp. (itNeptunia) and roots of Neptunia natans L. f. (Druce) (previously N. oleracea Lour.) under natural and laboratory conditions. When grown in its native marsh habitat, this unusual aquatic legume does not develop root hairs, the primary sites of rhizobial infection for most temperate legumes. Under natural conditions, the aquatic plant floats and develops nitrogen-fixing nodules at emergence of lateral roots on the primary root and on adventitious roots at stem nodes, but not from the stem itself. Cytological studies using various microscopies revealed that the mode of root infection involved an intercellular route of entry followed by an intracellular route of dissemination within nodule cells. After colonizing the root surface, the bacteria entered the primary root cortex through natural wounds caused by splitting of the epidermis and emergence of young lateral roots, and then stimulated early development of nodules at the base of such roots. The bacteria entered the nodule through pockets between separated host cells, then spread deeper in the nodule through a narrower intercellular route, and eventually evoked the formation of infection threads that penetrated host cells and spread throughout the nodule tissue. Bacteria were released from infection droplets at unwalled ends of infection threads, became enveloped by peribacteroid membranes, and transformed into enlarged bacteroids within symbiosomes. In older nodules, the bacteria within symbiosomes were embedded in an unusual, extensive fibrillar matrix. Cross-inoculation tests of 18 isolates of rhizobia from nodules of N. natans revealed a host specificity enabling effective nodulation of this aquatic legume, with lesser affinity for Medicago sativa and Ornithopus sp., and an inability to nodulate several other crop legume species. Acetylene reduction (N₂ fixation) activity was detected in nodules of N. natans growing in aquatic habitats under natural conditions in Southern India. These studies indicate that a specific group of Rhizobium sp. (Neptunia) occupies a unique ecological niche in aquatic environments by entering into a N₂-fixing root-nodule symbiosis with Neptunia natans.We thank J. Whallon for technical assistance, G. Truchet, J. Vasse, S. Wagener, J. Beaman, F. DeBruijn, F. Ewers, and A. Squartini for helpful comments, and N.N. Prasad and G. Birla for assistance in conducting field observations. This work was supported by the Michigan Agricultural Experiment Station and National Science Foundation grants DIR-8809640 and BIR-9120006 awarded to the MSU Center for Microbial Ecology. This study is dedicated to the memory of Dr. Joseph C. Burton, a friend and colleague who made many contributions to the study of the Rhizobiumlegume symbiosis.     

Impact of two fluorescent pseudomonads and an arbuscular mycorrhizal fungus on tomato plant growth,root architecture and P acquisition

The ability of fluorescent pseudomonads and arbuscular mycorrhizal fungi (AMF) to promote plant growth is well documented but knowledge of the impact of pseudomonad-mycorrhiza mixed inocula on root architecture is scanty. In the present work, growth and root architecture of tomato plants (Lycopersicon esculentum Mill. cv. Guadalete), inoculated or not with Pseudomonas fluorescens 92rk and P190r and/or the AMF Glomus mosseae BEG12, were evaluated by measuring shoot and root fresh weight and by analysing morphometric parameters of the root system. The influence of the microorganisms on phosphorus (P) acquisition was assayed as total P accumulated in leaves of plants inoculated or not with the three microorganisms. The two bacterial strains and the AMF, alone or in combination, promoted plant growth. P. fluorescens 92rk and G. mosseae BEG12 when co-inoculated had a synergistic effect on root fresh weight. Moreover, co-inoculation of the three microorganisms synergistically increased plant growth compared with singly inoculated plants. Both the fluorescent pseudomonads and the myco-symbiont, depending on the inoculum combination, strongly affected root architecture. P. fluorescens 92rk increased mycorrhizal colonization, suggesting that this strain is a mycorrhization helper bacterium. Finally, the bacterial strains and the AMF, alone or in combination, improved plant mineral nutrition by increasing leaf P content. These results support the potential use of fluorescent pseudomonads and AMF as mixed inoculants for tomato and suggest that improved tomato growth could be related to the increase in P acquisition.     

Bacterial effects on arbuscular mycorrhizal fungi and mycorrhiza development as influenced by the bacteria, fungi, and host plant

Bacterial strains from mycorrhizal roots (three belonging to Comamonadaceae and one to Oxalobacteraceae) and from non-mycorrhizal roots (two belonging to Comamonadaceae) of Medicago truncatula and two reference strains (Collimonas fungivorans Ter331 and Pseudomonas fluorescens C7R12) were tested for their effect on the in vitro saprophytic growth of Glomus mosseae BEG12 and on its colonization of M. truncatula roots. Only the Oxalobacteraceae strain, isolated from barrel medic mycorrhizal roots, and the reference strain P. fluorescens C7R12 promoted both the saprophytic growth and root colonization of G. mosseae BEG12, indicating that they acted as mycorrhiza helper bacteria. Greatest effects were achieved by P. fluorescens C7R12 and its influence on the saprophytic growth of G. mosseae was compared to that on Gigaspora rosea BEG9 to determine if the bacterial stimulation was fungal specific. This fungal specificity, together with plant specificity, was finally evaluated by comparing bacterial effects on arbuscular mycorrhizal symbiosis when each of the fungal species was inoculated to two different plant species (M. truncatula and Lycopersicon esculentum). The results obtained showed that promotion of saprophytic growth by P. fluorescens C7R12 was expressed in vitro towards G. mosseae but not towards G. rosea. Bacterial promotion of mycorhization was also expressed towards G. mosseae, but not G. rosea, in roots of M. truncatula and L. esculentum. Taken together, results indicated that enhancement of arbuscular mycorrhiza development was only induced by a limited number of bacteria, promotion by the most efficient bacterial strain being fungal and not plant specific.     

Different Ks values for hydrogen of methanogenic bacteria and sulfate reducing bacteria: An explanation for the apparent inhibition of methanogenesis by sulfate

Desulfovibrio vulgaris (Marburg) and Methanobrevibacter arboriphilus (AZ) are anaerobic sewage sludge bacteria which grow on H₂ plus sulfate and H₂ plus CO₂ as sole energy sources, respectively. Their apparent K_s values for H₂ were determined and found to be approximately 1 M for the sulfate reducing bacterium and 6 M for the methanogenic bacterium. In mixed cell suspensions of the two bacteria (adjusted to equal V _max) the rate of H₂ consumption by D. vulgaris was five times that of M. arboriphilus, when the hydrogen supply was rate limiting. The apparent inhibition of methanogenesis was of the same order as expected from the different K_s values for H₂. Difference in substrate affinities can thus account for the inhibition of methanogenesis from H₂ and CO₂ in sulfate rich environments, where the H₂ concentration is well below 5 M.     

Influence of nitrogen source on hydrogen generation by a photosynthetic bacterium

Axenic culture of photosynthetic purple sulphur bacterium, isolated from a paper mill effluent, grew best with ammonium chloride as nitrogen source. Cells grown with N₂ or ammonium chloride produced hydrogen with initial lag periods of 24 and 48 h, respectively. The maximum amount of hydrogen was evolved by cells grown with N₂ and with malate as electron donor.R. Lakshmi is with the Microbial Physiology Laboratory, Department of Botany, and H. Polasa is with the Department of Microbiology, Osmania University, Hyderabad 500 007, India.     

Sporomusa,a new genus of gram-negative anaerobic bacteria including Sporomusa sphaeroides spec. nov. and Sporomusa ovata spec. nov.

A new genus of strictly anaerobic, gram-negative, banana-shaped bacteria is described. Cells formed spores and were motile by means of up to 15 laterally inserted flagella. Nitrate or sulfate were not used as electron acceptor. Organic substrates that were fermented included N-methyl compounds, such as betaine, N,N-dimethylglycine and sarcosine, primary alcohols, hydroxy fatty acids, and 2,3-butanediol. In addition, molecular hydrogen and carbon dioxide were fermented to acetate. The latter was the characteristic fermentation product in general. During growth on betaine, trimethylamine was formed in addition. The degradation of N,N-dimethylglycine yielded acetate, monomethylamine, and trimethylamine. The presence of cytochrome b and of ubiquinone in the cells was shown. The deoxyribonuleic acid base composition of the strains was between 41.3 and 47.4 mol% guanine plus cytosine. The name Sporomusa is proposed for this new genus. On the basis of the DNA-DNA homology values obtained, the shape of the spores and some other properties, the isolated strains were assigned to two species. Names proposed: Sporomusa sphaeroides and Sporomusa ovata. The type species is S. sphaeroides and the type strains are strain E, DSM 2875 (S. sphaeroides) and strain H1, DSM 2662 (S. ovata).Dedicated to Prof. H. G. Schlegel on the occasion of his 60th birthday     

The effect of sulfur compounds on H<Subscript>2</Subscript> evolution/consumption reactions,mediated by various hydrogenases,in the purple sulfur bacterium, <Emphasis Type="Italic">Thiocapsa roseopersicina</Emphasis>

The influence of reduced sulfur compounds (including stored S⁰) on H₂ evolution/consumption reactions in the purple sulfur bacterium, Thiocapsa roseopersicina BBS, was studied using mutants containing only one of the three known [NiFe] hydrogenase enzymes: Hox, Hup or Hyn. The observed effects depended on the kind of hydrogenase involved. The mutant harbouring Hox hydrogenase was able to use S₂O₃²⁻, SO₃²⁻, S²⁻ and S⁰ as electron donors for light-dependent H₂ production. Dark H₂ evolution from organic substrates via Hox hydrogenase was inhibited by S⁰. Under light conditions, endogenous H₂ uptake by Hox or Hup hydrogenases was suppressed by S compounds. СО₂-dependent H₂ uptake by Hox hydrogenase in the light required the additional presence of S compounds, unlike the Hup-mediated process. Dark H₂ consumption via Hyn hydrogenase was connected to utilization of S⁰ as an electron acceptor and resulted in the accumulation of H₂S. In wild type BBS, with high levels of stored S⁰, dark H₂ production from organic substrates was significantly lower, but H₂S accumulation significantly higher, than in the mutant GB1121(Hox⁺). There is a possibility that H₂ produced via Hox hydrogenase is consumed by Hyn hydrogenase to reduce S⁰.     

Sulfhydryl modifying reagents inhibit Q A − oxidation in reaction centers from Rhodobacter sphaeroides and Capsulatus,but not Rhodopseudomonas viridis

The effects of various sulfhydryl-modifying reagents on reaction centers (RCs) from purple photosynthetic bacteria have been examined, with particular emphasis on the activity of the acceptor quinones, Q_A and Q_B, comprising the two electron gate. Mercurial reagents, especially p-chloromercuribenzenesulfonate (pCMBS), were effective in inhibiting Q_B function in RCs from Rhodobacter sphaeroides and Rb. capsulatus, but not in Rhodopseudomonas viridis. The inhibition was fully reversible by dialysis against dithiothreitol (DTT). The effect on Q_B function was not an apparent one mediated by an alteration in the redox potential of Q_A. N-ethylmaleimide (NEM) had no effect on any of the quinone functions, even at very high concentrations. Comparison of the X-ray structures of the RCs from Rb. sphaeroides and Rp. viridis and the known amino acid sequences for all three bacterial RCs suggest that a cysteine residue at position 108 in the L subunit of the Rhodobacter species is the most likely candidate for the site of action of the mercurial reagents. This was strongly supported by the absence of any effect of pCMBS on a site specific mutation of Rb. sphaeroides (L108CS) with residue L108 changed from cysteine to serine. These results imply a long distance (>20 Å) effect on the functioning of Q_B, perhaps involving a relatively gross structural alteration.     

Gas phase H<Subscript>2</Subscript>S product recovery in a packed bed bioreactor with immobilized sulfate-reducing bacteria

An N₂ strip gas was used in a packed bed sulfate-reducing bioreactor to recover the dissolved sulfide product and improve sulfate conversion. The highest volumetric productivity obtained was 261 mol H₂S m⁻³ d⁻¹. Lowering the initial pH of the medium from 7 to 6 increased the H₂S content of the strip gas from 3.6 to 5.8 mol%. The ratio of strip gas to liquid flow rates (G/L) was found be to a suitable basis for scaling the process. Calculations indicated that modest G/L values (<10²) were required to recover the residual dissolved sulfide in a downstream stripping column.     

Colonization behaviour of Pseudomonas fluorescens and Sinorhizobium meliloti in the alfalfa (Medicago sativa) rhizosphere

The colonization ability of Pseudomonas fluorescens F113rif in alfalfa rhizosphere and its interactions with the alfalfa microsymbiont Sinorhizobium meliloti EFB1 has been analyzed. Both strains efficiently colonize the alfalfa rhizosphere in gnotobiotic systems and soil microcosms. Colonization dynamics of F113rif on alfalfa were similar to other plant systems previously studied but it is displaced by S. meliloti EFB1, lowering its population by one order of magnitude in co-inoculation experiments. GFP tagged strains used to study the colonization patterns by both strains indicated that P. fluorescens F113rif did not colonize root hairs while S. meliloti EFB1 extensively colonized this niche. Inoculation of F113rif had a deleterious effect on plants grown in gnotobiotic systems, possibly because of the production of HCN and the high populations reached in these systems. This effect was reversed by co-inoculation. Pseudomonas fluorescens F113 derivatives with biocontrol and bioremediation abilities have been developed in recent years. The results obtained support the possibility of using this bacterium in conjunction with alfalfa for biocontrol or rhizoremediation technologies.     

Desulfacinum subterraneumsp. nov., a New Thermophilic Sulfate-Reducing Bacterium Isolated from a High-Temperature Oil Field

A new thermophilic sulfate-reducing bacterium isolated from the high-temperature White Tiger oil field (Vietnam) is described. Cells of the bacterium are oval (0.4–0.6 by 0.6–1.8 m), nonmotile, non-spore-forming, and gram-negative. Growth occurs at 45 to 65°C (with an optimum at 60°C) at NaCl concentrations of 0 to 50 g/l. In the course of sulfate reduction, the organism can utilize lactate, pyruvate, malate, fumarate, ethanol, salts of fatty acids (formate, acetate, propionate, butyrate, caproate, palmitate), yeast extract, alanine, serine, cysteine, and H₂+ CO₂(autotrophically). In addition to sulfate, the bacterium can use sulfite, thiosulfate, and elemental sulfur as electron acceptors. In the absence of electron acceptors, the bacterium can ferment pyruvate and yeast extract (a yet unrecognized capacity of sulfate reducers) with the formation of acetate and H₂. The G+C content of DNA is 60.8 mol %. The level of DNA–DNA hybridization of the isolate (strain 101^T) and Desulfacinum infernum(strain BG1^T) is as low as 34%. Analysis of the nucleotide sequence of 16S rDNA places strain 101^Tin the phylogenetic cluster of the Desulfacinumspecies within the sulfate reducer subdivision of the delta subclass of Proteobacteria. All these results allowed the bacterium studied to be described as a new species, Desulfacinum subterraneumsp. nov., with strain 101 as the type strain.     

Genetic diversity within and among populations of Shorea leprosula Miq. and Shorea parvifolia Dyer (Dipterocarpaceae) in Indonesia detected by AFLPs

The genetic diversity within and among populations of Shorea leprosula and Shorea parvifolia from Indonesia was investigated using amplified fragment length polymorphisms (AFLPs). The results indicated that S. leprosula is genetically more variable than S. parvifolia. At the population level, a higher level of genetic diversity was revealed for S. leprosula with a percentage of polymorphic loci (PPL_p) of 53.32% and an expected heterozygosity (H _ep) of 0.16 in comparison with S. parvifolia showing PPL_p of 51.79% and H _ep of 0.14. At the species level, S. leprosula showed PPL_s of 92.86% and H _es of 0.21, while S. parvifolia showed PPL_s of 85.71% and H _es of 0.21. Genetic differentiation (G _st) indicated that 25 and 31% of total genetic diversity in S. leprosula and S. parvifolia, respectively, were attributed to the differences among populations. An analysis of molecular variance (AMOVA) at two hierarchical levels exhibited that most genetic variation resided within populations with proportion of 70.2% for S. leprosula and 66.2% for S. parvifolia. The AMOVA at three hierarchical levels performed for S. leprosula and S. parvifolia together revealed that the genetic difference between the two species was remarkably higher with a proportion of 44.1% than the differences within and among populations (38.1 and 17.8%, respectively). The genetic differentiation between islands was significant for S. leprosula but not for S. parvifolia. The observed genetic diversity agreed with the life history traits of Shorea species. Highly differentiating individual AFLP markers were found for each species, which will serve as diagnostic markers for the identification of wood of different species, from different islands and regions.     

A new anaerobic bacterium,forming up to five endospores per cell —Anaerobacter polyendosporus gen. et spec. nov.

An obligately anaerobic sporeforming bacterium assigned to a new genus and species Anaerobacter polyendosporus gen. et spec. nov. is described. Characteristic features distinguishing the bacterium from known anaerobic sporeformers were variable cell shape, including spherical, the ability to form up to five endospores per cell, diffusive distribution of reserve polysaccharide throughout the cytoplasm, independence from growth factors. The eubacterial nature of the organism was revealed by its sensitivity to 1 mg/l of streptomycin, rifampicin, penicillin and to lysozyme. It belonged to Firmicutes by the type of cell wall structure. The cell wall consisted of one layer; the outer membrane was absent. The cells were not motile. The spores were spherical or oval, heat-resistant, contained dipicolinic acid and had typical endospore structure. Cortex, coats, spore coare, and in most cases exosporium could be distinguished. The bacterium fermented carbohydrates, but not amino acids. The products of fermentation included ethanol, acetate, lactate, butyrate, butanol, H₂ and CO₂. Sulfate or nitrate could not be used as electron acceptors, but nitrite was reduced to NH ₄ ⁺ in a dissimilatory process. The bacterium was capable of fixing N₂. The G + C content of the DNA was 29 mol %. The bacterium was isolated from meadow-gley soil.     

黄土丘陵区优势造林树种水分来源对季节性干旱的响应

黄土丘陵区处于季节性干旱生态脆弱地带,探明水分环境变化对区域优势造林树种水分来源的影响,对干旱区植物水分利用及其共生关系具有重要意义。以该地区广泛种植的沙棘(Hippophae rhamnoides)+油松(Pinus tabuliformis)、沙棘(H.rhamnoides)+刺槐(Robinia pseudoacacia)混交林(H_rP_t,H_rR_p)及沙棘(H_r)、油松(P_t)和刺槐(R_p)纯林为对象,测定其茎秆水、土壤水和降水稳定同位素组成,采用IsoSource模型量化水分来源,相似性比例指数(PS)定量分析共生植物间水分利用关系。结果表明：(1)旱季,同一树种纯林和混交林间吸水层位存在差异,纯林中主要利用0—100 cm土层土壤水,其中H_r和R_p对40—100 cm的利用率为46.8%和43.8%,P_t对0—40 cm为83.9%;混交林中则更倾向于利用40...     

Clostridium mayombei sp. nov., an H2/CO2 acetogenic bacterium from the gut of the African soil-feeding termite,Cubitermes speciosus

Clostridium mayombei sp. nov., a previously undescribed H₂-oxidizing CO₂-reducing acetogenic bacterium, was isolated from gut contents of the African soilfeeding termite, Cubitermes speciosus. Cells were anaerobic, Gram positive, catalase and oxidase negative, endospore-forming motile rods which measured 1×2 – 6 m and which had a DNA base composition of 25.6 mol% G+C (strain SFC-5). Optimum conditions for growth on H₂+CO₂ were at 33°C and pH 7.3, and under these conditions cells produced acetate according to the equation: 4 H₂+2 CO₂CH₃COOH+2 H₂O. Other substrates supporting good growth included carbohydrates (e.g. glucose, xylose, starch), sugar alcohols, and organic and amino acids, and with these substrates acetate was almost always the principle fermentation product. Comparative analysis of 16S rRNA nucleotide sequences confirmed that C. mayombei was closely related to various members of the genus Clostridium. However, morphological and physiological differences between C. mayombei and other homoacetogenic clostridia were deemed significant enough to warrant creation of a new taxon. Results are discussed in light of the diversity of H₂/CO₂ acetogens recently isolated from various termites, and in terms of the relative importance of H₂/CO₂ acetogenesis to termite nutrition.     

DMSO respiration by the anaerobic rumen bacterium Wolinella succinogenes

The anaerobic rumen bacterium Wolinella succinogenes was able to grow by respiration with dimethylsulphoxide (DMSO) as electron acceptor and formate or H₂ as electron donors. The growth yield amounted to 6.7 g and 6.4 g dry cells/mol DMSO with formate or H₂ as the donors, respectively. This suggested an ATP yield of about 0.7 mol ATP/mol DMSO. Cell homogenates and the membrane fraction contained DMSO reductase activity with a high K _m (43 mM) for DMSO. The electron transport from H₂ to DMSO in the membranes was inhibited by 2-(heptyl)-4-hydroxyquinoline N-oxide, indicating the participation of menaquinone. Formation of DMSO reductase activity occurred only during growth on DMSO, presence of other electron acceptors (fumarate, nitrate, nitrite, N₂O, and sulphur) repressed the DMSO reductase activity. DMSO can therefore be used by W. succinogenes as an acceptor for phosphorylative electron transport, but other electron acceptors are used preferentially.Abbreviations DMN 2,3-Dimethyl-1,4-naphthoquinone - DMNH ₂ Reduced DMN - DMS Dimethylsulphide (CH₃)₂S - DMSO Dimethylsulphoxide (CH₃)₂SO - HQNO 2-(Heptyl)-4-hydroxyquinoline-N-oxide - TMAO Trimethylamine-N-oxide - Y _s Growth yield for substrate S