Photoinduced electron transfer and cytochrome content in obligate aerobic phototrophic bacteria from genera Erythromicrobium,Sandaracinobacter, Erythromonas,Roseococcus and Erythrobacter

The photosynthetic apparatus and the electron carriers of seven species of five different genera of obligate aerobic phototrophic bacteria have been characterized by biochemical and biophysical techniques. A tetrahemic reaction center (RC) bound cytochrome (cyt) was found in Erythromonas (Em.) ursincola, Sandaracinobacter (S.) sibiricus and Roseococcus (R.) thiosulfatophilus, but not in Erythromicrobium (E.) ezovicum, Erythromicrobium ramosum, Erythromicrobium hydrolyticum and Erythrobacter (Eb.) litoralis. In none of the studied species, photochemical activity was observed under anaerobic conditions. Under aerobic conditions, the photoinduced cyclic electron transfer involves a soluble c-type cyt for the seven species. The cyt content of soluble and membrane fractions is highly dependent upon the species. The Erythromicrobium species (E. ezovicum, E. ramosum and E. hydrolyticum) contains a major soluble cyt while the other species possess several soluble cyts, up to four in the case of Eb. litoralis. These cyts have been characterized in terms of midpoint potential and apparent molecular mass. The presence of cyt bc₁ complexes has been clearly detected in Eb. litoralis, E. hydrolyticum, E. ezovicum and E. ramosum. These last three species also contain a high midpoint potential (350 mV) membrane-bound cyt c of unknown function.     

Dynamics of aerobic anoxygenic phototrophic bacteria in the East China Sea

Aerobic anoxygenic phototrophic bacteria (AAPB) are a group of heterotrophic bacteria capable of photosynthesis. The dynamics of AAPB in the East China Sea, a typical marginal sea characterized by diverse physical-chemical and ecological conditions, were investigated from April 2002 to September 2003. The results showed that the abundance of AAPB varied from 0.16 to 7.9 x 10(4) cells mL(-1) and the percentage of AAPB (AAPB%) in the total heterotrophic bacterial abundance varied from 0.5% to 11.6% over a gradient of environmental conditions. The abundance of AAPB and AAPB% was higher in coastal and continental shelf waters than in oceanic waters. An interesting seasonal pattern was observed in the Yangtze River estuary: the abundance of AAPB was highest in summer and lowest in winter; however, AAPB% was higher in winter than in the other seasons. Throughout the investigation period, variation of AAPB abundance with temperature was much less than that of nonAAPB abundance, suggesting that low temperature was not a limiting factor for AAPB in this case. Close correlation between AAPB and chlorophyll a was observed in each season, suggesting that dependence of AAPB on dissolved organic carbon produced by phytoplankton (PDOC) may be one key factor controlling AAPB distribution.     

On the natural selection and evolution of the aerobic phototrophic bacteria

This contribution gives a brief survey of the short history since the discovery of the aerobic phototrophic bacteria to focus on a general evolutionary scenario. Most of the citations are of reviews that have covered the earlier literature and to which the reader is directed at appropriate places in the following text. The data summarized in these reviews are supplemented with information from recent or otherwise key primary publications in order to support a synthesis that addresses vexing questions about bacteria containing photosynthetic pigment-protein complexes, but which are incapable of growth with light as the sole, or even the major source of energy. This revised version was published online in June 2006 with corrections to the Cover Date.     

Bacteriochlorophyll-protein complexes of aerobic bacteria,Erythrobacter longus and Erythrobacter species OCh 114

Bacteriochlorophyll(Bchl)-protein complexes were isolated from obligate aerobic bacteria, Erythrobacter longus and Erythrobacter species OCh 114. The apparent molecular weights, absorption spectra and polypeptide compositions of the light-harvesting complexes were, in general, similar to those of the light-harvesting Bchl-protein complexes of purple photosynthetic bacteria. The reaction center complexes of these bacteria also showed similar properties to those of the purple bacteria except for slightly altered polypeptides. However, the following characteristic features of the light-harvesting systems were found in these aerobic bacteria. Major carotenoids were not bound to the Bchl-protein complex in E. longus. In Erythrobacter sp. OCh 114, a new type of Bchl-protein complex which showed a single absorption band in the near infrared region at 806 nm was obtained. The reaction center of strain OCh 114 was associated with a c-type cytochrome.Abbreviations Bchl bacteriochlorophyll a - RC reaction center - SDS sodium dodecylsulfate - PAGE polyacrylamide gel electrophoresis     

Quinones of phototrophic purple bacteria

Abstract The quinone composition of the recognized species of the phototrophic purple nonsulfur bacteria, the Ectothiorhodospiraceae, and some Chromatiaceae species has been determined. Altogether more than 50 strains of 33 species have been investigated. Some of the purple nonsulfur bacteria have Q-10 as sole quinone component, while others have Q-10, Q-9, or Q-8, respectively, together with menaquinones of the same isoprenoid chain length as the major components. Rhodoquinone is present in Rhodospirillum rubrum and Rhodospirillum photometricum . The Ectothiorhodospira species have either Q-8 and MK-8, like the Chromatiaceae species, or Q-7 and MK-7 as the major components.     

Primary photochemistry of reaction centers from the photosynthetic purple bacteria

Photosynthetic organisms transform the energy of sunlight into chemical potential in a specialized membrane-bound pigment-protein complex called the reaction center. Following light activation, the reaction center produces a charge-separated state consisting of an oxidized electron donor molecule and a reduced electron acceptor molecule. This primary photochemical process, which occurs via a series of rapid electron transfer steps, is complete within a nanosecond of photon absorption. Recent structural data on reaction centers of photosynthetic bacteria, combined with results from a large variety of photochemical measurements have expanded our understanding of how efficient charge separation occurs in the reaction center, and have changed many of the outstanding questions.Abbreviations BChl bacteriochlorophyll - P a dimer of BChl molecules - BPh bacteriopheophytin - Q_A and Q_B quinone molecules - L, M and H light, medium and heavy polypeptides of the reaction center     

Mechanisms of excitation trapping in reaction centers of purple bacteria

The contradiction between two groups of experimental data, which fails to be resolved within the framework of the widely accepted model of excitation migration and trapping (at least in case of purple bacteria), is discussed in the introduction to this review. Three directions of studies intended to resolve this conflict are reviewed in the three further sections: II. Exciton models; III. Water-polarization (water-latch) mechanism of excitation trapping; IV. Quantum-mechanical models. The maximum efficiency of these models in resolving the contradiction mentioned above was assessed. The advantages and disadvantages of the mechanisms described in sections II, III, and IV are discussed in the last section of this review. It is concluded that none of these mechanisms taken alone is able to solve this problem. Therefore, the fundamental problem of the primary excitation conversion in reaction centers remains unsolved and requires additional experimental research.     

Vertical distribution and characterization of aerobic phototrophic bacteria at the Juan de Fuca Ridge in the Pacific Ocean

The vertical distribution of culturable anoxygenic phototrophic bacteria was investigated at five sites at or near the Juan de Fuca Ridge in the Pacific Ocean. Twelve similar strains of obligately aerobic phototrophic bacteria were isolated in pure culture, from depths ranging from 500 to 2,379 m below the surface. These strains appear morphologically, physiologically, biochemically, and phylogenetically similar to Citromicrobium bathyomarinum strain JF-1, a bacterium previously isolated from hydrothermal vent plume waters. Only one aerobic phototrophic strain was isolated from surface waters. This strain is morphologically and physiologically distinct from the strains isolated at deeper sampling locations, and phylogenetic analysis indicates that it is most closely related to the genus Erythrobacter. Phototrophs were cultivated from three water casts taken above vents but not from two casts taken away from active vent sites. No culturable anaerobic anoxygenic phototrophs were detected. The photosynthetic apparatus was investigated in strain JF-1 and contains light-harvesting I and reaction center complexes, which are functional under aerobic conditions.     

Taxonomic differences shape the responses of freshwater aerobic anoxygenic phototrophic bacterial communities to light and predation

Aerobic anoxygenic phototrophic (AAP) bacteria are a phylogenetically diverse and ubiquitous group of prokaryotes that use organic matter but can harvest light using bacteriochlorophyll a. Although the factors regulating AAP ecology have long been investigated through field surveys, the few available experimental studies have considered AAPs as a group, thus disregarding the potential differential responses between taxonomically distinct AAP assemblages. Here, we used sequencing of the pufM gene to describe the diversity of AAPs in 10 environmentally distinct temperate lakes, and to investigate the taxonomic responses of AAP communities in these lakes when subjected to similar experimental manipulations of light and predator removal. The studied communities were clearly dominated by Limnohabitans AAP but presented a clear taxonomic segregation between lakes presumably driven by local conditions, which was maintained after experimental manipulations. Predation reduction (but not light exposure) caused significant compositional shifts across most assemblages, but the magnitude of these changes could not be clearly related to changes in bulk AAP abundances or taxonomic richness of AAP assemblages during experiments. Only a few operational taxonomic units, which differed taxonomically between lakes, were found to respond positively during experimental treatments. Our results highlight that different freshwater AAP communities respond differently to similar control mechanisms, highlighting that in‐depth knowledge on AAP diversity is essential to understand the ecology and potential role of these photoheterotrophs.     

黄河滩地和稻田土中铁还原菌、不产氧光合细菌分布机制

【目的】探讨沿黄流域土壤中铁还原菌(ferric reducing bacteria, Fe RB)、不产氧光合细菌(anoxygenic phototrophic bacteria, An PB)的分布机制。【方法】以沿黄流域(原阳段)为研究对象,采集黄河滩地和稻田土样,利用16Sr RNA基因高通量测序和实时荧光定量分析技术,结合统计学分析,揭示Fe RB、An PB菌群结构、丰度和主要环境影响因子。【结果】二者中的优势Fe RB在科(属)水平为Hydrogenophilaceae(Thiobacillus)、 Bacillaceae(Bacillus)、 Clostridiaceae、Rhodobactereace(Rhodobacter)、 Geobacteraceae(Geobacter),优势An PB为Rhodobactereace(Rhodobacter)、 Chloroflexaceae(Chloronema)、 Acetobacteraceae(Roseomonas)。An PB中Rhodobacteraceae与Fe RB中Bacillaceae、 Clostri...     

玛珥湖好氧不产氧光合细菌pufM基因DNA和mRNA的定量及多样性分析

【目的】湖光岩玛珥湖是一类特殊的火山口湖,它完全封闭,地质年代久远,尚未受人类活动的剧烈影响,孕育着丰富而特殊的微生物种群。好氧不产氧光合细菌(AAPB)是以其在有氧情况下能行使光合功能而定义的一类专性异养细菌,其生理生态特征独特,进化年代久远,在水生生态系统的上层水体中广泛分布。目前,AAPB在玛珥湖水体中是否有分布仍是未知。【方法】构建和比较夏季湖水1 m、5 m、12 m三个水层的总DNA和总RNA的AAPB光合中心合成的关键基因pufM的6个克隆文库,并结合定量PCR技术,分析了不同水层AAPB的分布、系统发育多样性及其在总细菌中的比重。【结果】6个文库覆盖率和稀释曲线显示样本初步揭示了各水层优势AAPB类群的多样性。BLAST核苷酸同源性介于80%93%;多样性指数表明,湖光岩表层和底层多样性相当,中间层最低,总RNA的多样性高于总DNA。系统发育分析结果表明,OTU21 24所含的序列(占总序列的49.43%)与β-变形细菌的进化距离最接近,是湖光岩玛珥湖的优势AAPB菌群。定量PCR结果显示1 m水层中AAPB在总细菌中的比重最高,可达38.06%;而5 m水层中AAPB所占的比重最低,仅为0.85%;12 m为9.54%。【结论】湖光岩玛珥湖孕育着丰富而多样的AAPB类群。     

Real-time PCR for quantification of aerobic anoxygenic phototrophic bacteria based on pufM gene in marine environment

The abundance of aerobic anoxygenic phototrophic bacteria (AAPB), a new functional group that plays important roles in marine carbon cycling, is determined frequently by infrared epifluorescence microscopic analysis (IREM) or high-performance liquid chromatography (HPLC) based on detecting BChl a (bacteriochlorophyll a) fluorescence signal at 880 nm. Unfortunately, the fluorescence signal is often influenced by environmental variables and physiological state of cell. Here we developed a real-time quantitative PCR (qPCR) assay based on pufM gene to specifically quantify AAPB in marine environments. High specificity and sensitivity for estimation of AAPB abundance were revealed by analysis of amplification products, melting curves and target sequences. The phylogenetic tree indicated that this primer set is suitable for a wide genetic diversity of AAPB, including α-3, α-4 Proteobacteria and clones of unclear taxonomic position. In contrast, no amplicon was obtained from green non-sulphur bacteria and oxygenic phototrophic bacteria such as Cyanobacterial genomic DNA. The melting behavior could indicate predominant phenotypes in AAPB community in addition to validating the products of qPCR. The AAPB was estimated to range from 1.3 × 10⁴ cell/ml to 3.4 × 10⁵ cell/ml in our 10 tested water samples by this qPCR assay. Further investigations on the abundance distribution of AAPB in marine environments using the qPCR assay may provide new insight into their ecological functions.     

帕米尔高原可培养需氧冷适应细菌及古菌多样性

【目的】帕米尔高原是贫营养、高辐射、干燥的冷环境,可能蕴藏丰富的冷适应微生物资源。本研究基于分离培养技术,探究帕米尔高原不同海拔梯度及不同培养条件下的冷适应微生物多样性。【方法】针对采集自帕米尔高原1000-2000m、2000-3000m、3000-4000m和4 000-5 000 m四个海拔梯度的土壤样品,选用TSA和R2A两种培养基于4°C进行冷适应细菌的分离培养,NOM和F6两种培养基于4°C和15°C进行冷适应古菌的分离培养。根据16S rRNA基因序列同源性对分离菌株进行鉴定,分析不同海拔梯度和不同培养条件的物种多样性及之间的差异性。【结果】本研究从帕米尔高原共分离得到419株需氧原核微生物,16S rRNA基因测序鉴定结果表明,分离菌株隶属于2个域、5个门、8个纲、18个目、28个科、49个属及118个种,为115种冷适应细菌及3种古菌。冷适应细菌中,γ-变形菌纲(Gammaproteobacteria)为第一优势纲,放线菌纲(Actinobacteria)中物种多样性最为丰富。高海拔地区的冷适应细菌物种多样性存在差异,海拔1 000-4 000 m的优势菌群均为γ-变...     

Leucine incorporation by aerobic anoxygenic phototrophic bacteria in the Delaware estuary

Aerobic anoxygenic phototrophic (AAP) bacteria are well known to be abundant in estuaries, coastal regions and in the open ocean, but little is known about their activity in any aquatic ecosystem. To explore the activity of AAP bacteria in the Delaware estuary and coastal waters, single-cell ³H-leucine incorporation by these bacteria was examined with a new approach that combines infrared epifluorescence microscopy and microautoradiography. The approach was used on samples from the Delaware coast from August through December and on transects through the Delaware estuary in August and November 2011. The percent of active AAP bacteria was up to twofold higher than the percentage of active cells in the rest of the bacterial community in the estuary. Likewise, the silver grain area around active AAP bacteria in microautoradiography preparations was larger than the area around cells in the rest of the bacterial community, indicating higher rates of leucine consumption by AAP bacteria. The cell size of AAP bacteria was 50% bigger than the size of other bacteria, about the same difference on average as measured for activity. The abundance of AAP bacteria was negatively correlated and their activity positively correlated with light availability in the water column, although light did not affect ³H-leucine incorporation in light–dark experiments. Our results suggest that AAP bacteria are bigger and more active than other bacteria, and likely contribute more to organic carbon fluxes than indicated by their abundance.     

Anoxygenic phototrophic bacteria of the high-altitude meromictic Lake Gek-Gel,Azerbaijan

The anoxygenic phototrophic bacterial community of the high-altitude meromictic Lake Gek-Gel (Azerbaijan) was investigated in September 2003. The highest concentration of bacteriochlorophyll e (48 μg/l) was detected at a depth of 30 m; the peak of bacteriochlorophyll a (4.5 μg/l) occurred at 29 m. Phylogenetic analysis revealed that brown-colored green sulfur bacteria Chlorobium phaeobacteroides predominated in the lake. Nonsulfur purple bacteria phylogenetically close to Blastochloris sulfoviridis were found in insignificant amounts; these organisms have not been previously reported in Lake Gek-Gel.     

Purification and properties of 1-phosphofructokinase from Escherichia coli

Abstract The thermophilic facultatively phototrophic green bacterium Chloroflexus aurantiacus strain Ok-70-fl was shown to possess sulfide-repressed hydrogenase activity. Biosynthesis of the enzyme was severely repressed by S²⁻ (5.7 mM) and stimulated specifically by Ni²⁺ and by molecular hydrogen. The hydrogenase was shown to be localized in the cytoplasmic membrane and could be solubilized from the latter by the detergent Triton X-100 in a state forming one enzymatically active band ( M _r 170 × 10³) in polyacrylamide gels. In the membraneous state, the hydrogenase had its maximal activity at 73°C and was active with methyl viologen, methylene blue, menadione and flavins, but not with NAD or NADP as electron acceptors. Solubilization of the enzyme with Triton X-100 resulted in a drastic increase in the FAD/FMN-linked activity.     

In vivo states and functions of carotenoids in an aerobic photosynthetic bacterium,Erythrobacter longus

In vivo states and functions of carotenoids in the membranes and the isolated RC-B865 pigment-protein complexes from an aerobic photosynthetic bacterium, Erythrobacter longus, are investigated by means of fluorescence excitation and resonance Raman (RR) spectra. Erythroxanthin sulfate, a dominant carotenoid species in the membranes (>70%), is found not to transfer the absorbed light energy to bacteriochlorophyll (Bchl), and its RR spectra are similar between the in vivo and in vitro states. These observations indicate that erythroxanthin sulfate does not interact with either Bchl or proteins in the membranes, and suggest that its function may be limited to photoprotection by quenching the harmful singlet oxygen. On the other hand, two other carotenoid species contained in the isolated RC-B865 complexes, zeaxanthin and bacteriorubixanthinal, have a high efficiency of energy transfer to Bchl (88±5%). The RR spectra of these two carotenoids, each of which can be selectively obtained by choosing the excitation wavelength, show some characteristics of interactions with proteins or Bchl.Abbreviations Bchl bacteriochlorophyll a - FWHM full width at half maximum - PAGE polyacrylamide gel electrophoresis - RC reaction center - RR resonance Raman - SDS sodium dodecyl sulfate     

Selection and characterization of aerobic bacteria capable of degrading commercial mixtures of low-ethoxylated nonylphenols

Aims:  Isolation and characterization of new bacterial strains capable of degrading nonylphenol ethoxylates (NP n EO) with a low ethoxylation degree, which are particularly recalcitrant to biodegradation.
Methods and Results:  Seven aerobic bacterial strains were isolated from activated sludges derived from an Italian plant receiving NP n EO-contaminated wastewaters after enrichment with a low-ethoxylated NP n EO mixture. On the basis of 16S rDNA sequence, the strains were positioned into five genera: Ochrobactrum , Castellaniella , Variovorax , Pseudomonas and Psychrobacter . Their degradation capabilities have been evaluated on two commercial mixtures, i.e. Igepal CO-210 and Igepal CO-520, the former rich in low ethoxylated congeners and the latter containing a broader spectrum of NP n EO, and on 4- n -nonylphenol (NP). The strains degraded Igepal CO-210, Igepal CO-520 and 4- n -NP all applied at the initial concentration of 100 mg l⁻¹, by 35–75%, 35–90% and 15–25%, respectively, after 25 days of incubation.
Conclusions:  Some of the isolated strains, in particular the Pseudomonas strains BCb12/1 and BCb12/3, showed interesting degradation capabilities towards low ethoxylated NP n EO congeners maintaining high cell vitality.
Significance and Impact of the Study:  Increased knowledge of bacteria involved in NP n EO degradation and the possibility of using the isolated strains in tailored process for a tertiary biological treatment of effluents of wastewater treatment plants.     

Kinetic study of PF and CarT states in the LM subunit purified from the wild-type Rhodobacter sphaeroides reaction centers

The kinetics of absorbance changes related to the charge-separated state, P^F, and to the formation and decay of the carotenoid triplet state (Car^T) were studied in the LM reaction center subunit isolated from a wild-type strain of the purple bacterium Rhodobacter sphaeroides (strain Y). The P^F lifetime is lengthened (20±1.5 ns) in the LM complex as compared to the intact reaction centers (11±1 ns). The yield of the carotenoid triplet formation is higher (0.28±0.01) in the LM complex than in native reaction centers. We interpret our results in terms of perturbations of a first-order reaction connecting the singlet and the triplet state of the radical-pair state. Our results, together with those of a recent work (Agalidis, I., Nuijs, A.M. and Reiss-Husson, F. (1987) Biochim. Biophys. Acta (in press)) are consistent with a high I to Q_A electron transfer rate in this LM subunit, which is metal-depleted.The LM complex is considerably more sensitive than the reaction centers to photooxidative damage in the presence of oxygen. This is not readily accounted for simply by the higher carotenoid triplet yield, and may suggest a greater accessibility of the internal structures in the absence of the H-subunit.The lifetime of the carotenoid triplet decay (6.4±0.3 s) in the LM subunit is unchanged compared to the native reaction centers.Abbreviations BChl bacteriochlorophyll - Bph bacteriopheophytin - Car carotenoid - Chl chlorophyll - cyt cytochrome - L, M and H subunits light, medium and heavy subunits of the reaction center complex - P^R triplet electronic state of the primary electron donor - P; Q_A the first stable electron acceptor, a bound quinone - RC reaction center - LDAO lauryldimethylamine N-oxide - SDS sodium dodecyl sulfate - UQ ubiquinone This paper is published in our new format. All future authors are requested to follow our new instructions (see Photosynthesis Research 10:519–526, 1986)—Editor.     

The effect of the pH level on the communities of anoxygenic phototrophic bacteria of soda lakes of the southeastern Transbaikal Region

The effect of pH on the structure of the communities of anoxygenic phototrophic bacteria (APB) was studied under laboratory conditions. Samples of natural APB communities were inoculated into media that differed in pH values, which were 7, 9.5, or 10.5. The structure of the APB communities in the obtained enrichment cultures at all pH values depended also on the mineralization levels of the media, which were the same as in the lakes from which samples were taken. The same dependence of the community structure on salinity was observed as in the case of the natural communities that had been described previously. APB were most diverse in the enrichment cultures grown at pH 9.5. The shift of the pH to either neutral or extremely alkaline values restricted the species diversity within the APB community, resulting in marked predominance of the most adapted forms. It was shown that the status of Ectothiorhodospira species within the community could serve not only as an indicator of salinity but also as an indicator of pH in soda lakes with a water mineralization of higher than 5 g/l. The statuses of various APB groups in the community as dependent on pH and salinity are discussed, as well as possible changes in these statuses due to changes in the water level and other environmental parameters in the studied lakes.