Effects of glycine on cell growth and pigment biosynthesis in Rhodobacter azotoformans.

The effect of exogenous glycine (a precursor for the biosynthesis of bacteriochlorophyll) on the cell growth and photopigment accumulation was investigated in phototrophic growing Rhodobacter azotoformans 134K20. The growth rate and the biomass of strain 134K20 were significantly inhibited by glycine addition when ammonium sulfate or glutamate were used as nitrogen sources and acetate or succinate as carbon sources. A characteristic absorption maximum at approximately 423 nm was present in the absorption spectra of glutamate cultures while it was absent by the addition of high-concentration glycine of 15 mM. The component account for the 423 nm peak was eventually identified as magnesium protoporphyrin IX monomethyl ester, a precursor of bacteriochlorophyll a (BChl a). Comparative analysis of pigment composition revealed that the amount of BChl a precursors was significantly decreased by the addition of 15-mM glycine while the BChl a accumulation was increased. Moreover, glycine changed the carotenoid compositions and stimulated the accumulation of spheroidene. The A850 /A875 in the growth-inhibited cultures was increased, indicating an increased level of the light-harvesting complex 2 compared to the reaction center. The exogenous glycine possibly played an important regulation role in photosynthesis of purple bacteria.

Expression and purification of an ArsM-elastin-like polypeptide fusion and its enzymatic properties.

Enzymes could act as a useful tool for environmental bioremediation. Arsenic (As) biomethylation, which can convert highly toxic arsenite [As(III)] into low-toxic volatile trimethylarsine, is considered to be an effective strategy for As removal from contaminated environments. As(III) S-adenosylmethyltransferase (ArsM) is a key enzyme for As methylation; its properties and preparation are crucial for its wide application. Currently, ArsM is usually purified as a His-tag fusion protein restricting widespread use due to high costs. In this study, to greatly reduce the cost and simplify the ArsM preparation process, an Elastin-like polypeptide (ELP) tag was introduced to construct an engineered Escherichia coli (ArsM-ELP). Consequently, a cost-effective and simple non-chromatographic purification approach could be used for ArsM purification. The enzymatic properties of ArsM-ELP were systematically investigated. The results showed that the As methylation rate of purified ArsM-ELP (> 35.49%) was higher than that of E. coli (ArsM-ELP) (> 10.39%) when exposed to 25 µmol/L and 100 µmol/L As(III), respectively. The purified ArsM-ELP was obtained after three round inverse transition cycling treatment in 2.0 mol/L NaCl at 32 °C for 10 min with the yield reaching more than 9.6% of the total protein. The optimal reaction temperature, pH, and time of ArsM-ELP were 30 °C, 7.5 and 30 min, respectively. The enzyme activity was maintained at over 50% at 45 °C for 12 h. The enzyme specific activity was 438.8 ± 2.1 U/µmol. ArsM-ELP had high selectivity for As(III). 2-Mercaptoethanol could promote enzyme activity, whereas SDS, EDTA, Fe2+, and Cu2+ inhibited enzyme activity, and Mg2+, Zn2+, Ca2+, and K+ had no significant effects on it.

Characterization of recombinant E. coli expressing arsR from Rhodopseudomonas palustris CGA009 that displays highly selective arsenic adsorption.

Innovative methods to lower arsenic (As) exposure are sought. The As regulatory protein (ArsR) is reported of having high affinity and specificity to arsenite [As(III)]. Rhodopseudomonas palustris CGA009 is a good model organism for studying As detoxification due to at least three ars operons and four diverse arsRRP1-4 on the genome. In this study, four Escherichia coli harboring arsRRP1-4 derived from CGA009 were engineered and tested regarding their As resistance. The results showed that E. coli (arsRRP2) displayed robust As(III) resistance, and its growth inhibition rate was only 2.9% when exposed to 3.0 mmol/L As(III). At pH 7.0, E. coli (arsRRP2) showed an enhanced As adsorption capacity. As(III) (2.32 mg/g (dry weight, dw)) and 1.47 mg/g arsenate [As(V)] was adsorbed representing a 4.2-fold and 1.3-fold increase respectively compared to the control strain. The adsorption process was well fitted to Langmuir isothermal mode. E. coli (arsRRP2) (1.0~12.0 g/L) could remove 30.3~82.2% of As (III) when exposed to 10 µg/L As(III). No increase in absorption to copper(II), zinc(II), chromium(III), and lead(II) could be detected. Our studies revealed that arsRRP1-4 from CGA009 could confer As(III) resistance; E. coli (arsRRP2) displayed the highest As resistance, selectivity, and adsorption capacity within a wider pH (5.0~9.0) and salinity (0~15.0 g/L NaCl) range, especially important as it could remove As(III) from low concentration As-containing water.

Nitrogen transformation under different dissolved oxygen levels by the anoxygenic phototrophic bacterium Marichromatium gracile.

Marichromatium gracile: YL28 (M. gracile YL28) is an anoxygenic phototrophic bacterial strain that utilizes ammonia, nitrate, or nitrite as its sole nitrogen source during growth. In this study, we investigated the removal and transformation of ammonium, nitrate, and nitrite by M. gracile YL28 grown in a combinatorial culture system of sodium acetate-ammonium, sodium acetate-nitrate and sodium acetate-nitrite in response to different initial dissolved oxygen (DO) levels. In the sodium acetate-ammonium system under aerobic conditions (initial DO = 7.20-7.25 mg/L), we detected a continuous accumulation of nitrate and nitrite. However, under semi-anaerobic conditions (initial DO = 4.08-4.26 mg/L), we observed a temporary accumulation of nitrate and nitrite. Interestingly, under anaerobic conditions (initial DO = 0.36-0.67 mg/L), there was little accumulation of nitrate and nitrite, but an increase in nitrous oxide production. In the sodium acetate-nitrite system, nitrite levels declined slightly under aerobic conditions, and nitrite was completely removed under semi-anaerobic and anaerobic conditions. In addition, M. gracile YL28 was able to grow using nitrite as the sole nitrogen source in situations when nitrogen gas produced by denitrification was eliminated. Taken together, the data indicate that M. gracile YL28 performs simultaneous heterotrophic nitrification and denitrification at low-DO levels and uses nitrite as the sole nitrogen source for growth. Our study is the first to demonstrate that anoxygenic phototrophic bacteria perform heterotrophic ammonia-oxidization and denitrification under anaerobic conditions.

Selective repression of light harvesting complex 2 formation in Rhodobacter azotoformans by light under semiaerobic conditions.

Photosystem formation in anaerobic anoxygenic phototrophic bacteria (APB) is repressed by oxygen but is de-repressed when oxygen tension decreases. Under semiaerobic conditions, the synthesis of photopigments and pigment protein complexes in Rhodobacter (Rba.) sphaeroides are repressed by light. AppA, a blue-light receptor, mediates this regulation. In the present study, it was showed that the synthesis of bacteriochlorophyll, carotenoid, and pigment protein complexes in Rba. azotoformans 134K20 was significantly repressed by oxygen. Oxygen exposure also led to a conversion of spheroidene to spheroidenone. In semiaerobically growing cells, light irradiation resulted in a decrease in the formation of photosystem, and blue light was found to be the most effective light source. Blue light reduced the contents of bacteriochlorophyll and carotenoid slightly, but had negligible effects on light harvesting complex (LH) 1 content, whereas the content of LH2 was significantly decreased indicating that blue light selectively repressed the synthesis of LH2 in semiaerobically growing 134K20. It was concluded that, similar to Rba. sphaeroides, a blue light receptor presented in strain 134K20 played important roles in its light-dependent repression. A possible mechanism involved in controlling the differential inhibitory of blue light on the synthesis of photosystem was discussed.

[Construction and Characterization of B850-Only LH2 Energy Transfer System in Purple Bacteria].

To seek microscopic molecular mechanism of energy transfer and complex reconstitution in the photosynthesis, the conditions for construction of B850-only peripheral light-harvesting complex (LH2) and their properties were investigated using absorption, fluorescence spectroscopy, molecular sieve chromatography, ultrafiltration and sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) from the purple bacteria. The results indicated that bacteriochlorophylls (BChl) of B800 incubated in 10 mmo · L(-1) Tris-HCl (pH 8.0) buffer are selectively released from their binding sites of LH2 of Rhodobacter azotoformans (A-LH2) by 0.08% (W/V) SDS. B850-only A-LH2 was constructed after removing free BChl mixing with 10% methyl alcohol by ultrafiltration. B850 BChl was released after A-LH2 was incubated for 240 min in dark at room temperature (RT). While BChl of B800 incubated in pH 1.9 buffer were selectively released from their binding sites of LH2 of Rhodopseudomonas palustris (P-LH2). The authors acquired two components using molecular sieve chromatography. Free BChl of one component was not removed and self-assembled to P-LH2. The other removed free BChl and B850-only P-LH2 was constructed. B850 unchanged after P-LH2 was incubated. P-LH2 α and ß subunits have different molecular weights, but those of A-LH2 are in the contrary. It is concluded that B850-only P-LH2 is more stable than A-LH2. The enigmatic split of the B800 absorption band was not observed in these LH2, but we acquired two kinds of B800-released LH2 from Rhodopseudomonas palustris. The authors' results may provide a new light to separate homogeneous Apoprotein LH2.

A unique low light adaptation mechanism in Rhodobacter azotoformans.

In this paper, we reported for the first time that Rhodobacter azotoformans was capable of synthesizing a spectral variant of peripheral light-harvesting complex (LH3), besides a high light form (LH2), in response to low light intensity. Carotenoid components in these complexes were analyzed by absorption spectra, high-pressure liquid chromatography and mass spectroscopy analysis. Only spheroidene carotenoid was detected in LH2, while LH3 possessed three kinds of carotenoids, spheroidene, spirilloxanthin, and anhydrorhodovibrin. The spirilloxanthin and anhydrorhodovibrin predominated in LH3 and were rarely found in Rhodobacter species. Carotenoid-to-bacteriochlorophyll energy transfer efficiency in LH3 increased by 4% compared to that in LH2. Raman spectroscopic properties of carotenoids in both complexes supported the view that carotenoids altered their planar configuration to a distorted form by interaction with protein matrix in response to low light conditions. In conclusion, the low light adaptation mechanism of Rba. azotoformans involved regulating the synthesis of LH3 and additional carotenoids as well as the configuration change of incorporated carotenoids.

Pigments accumulation via light and oxygen in Rhodobacter capsulatus strain XJ-1 isolated from saline soil.

A Rhodobacter capsulatus strain, designated XJ-1, isolated from saline soil, accumulated almost only one kind of bacteriochlorophyll a anaerobically in the light, aerobically in the light and dark, and the relative contents of the bacteriochlorophyll a were 44.61, 74.89, and 77.53% of the total pigments, respectively. A new purple pigment appeared only in aerobic-light grown cells, exhibited absorption maxima at 355, 389, 520, 621, and 755 nm, especially distinctly unusual peak at 621 nm, whereas vanished in anaerobic-light and in aerobic-dark culture. Spheroidene and OH-spheroidene predominated in anaerobic phototrophic cultures. Spheroidenone was the sole carotenoid when exposed to both light and oxygen. The second keto-carotenoids, OH-spheroidenone, presented only in aerobic-dark culture in addition to spheroidenone. Strain XJ-1 would be a good model organism for the further illustration of the regulation of bacteriochlorophyll biosynthesis gene expression in response to unique habitat.

[Influence of iron on siderophore and photosynthetic pigments biosynthesis by siderophore-producing Rhodopesudomonnas palustris].

OBJECTIVE: To explore the regulation of iron on siderophore production, cell growth and photosynthetic pigments biosynthesis by siderophore-producing anoxygenic phototrophic bacteria. METHODS: Siderophore production was determined using Chrome Azurol S (CAS) assay. The siderophore types were determined by Arnow method, Csaky test and Shenker test. The compositions and contents of photosynthetic pigments were determined by spectrophotometry and HPLC analysis. RESULTS: Rhodopseudomonas palustris (Rps. palustris) CQV97 was capable of producing hydroxamate-type of siderophore. Siderophore production reached the highest yield in the absence of ferric chloride. With increasing ferric chloride concentrations, the lag phase of cell growth was shortened, and the cell growth rate, final biomass and the total amounts of carotenoid and bacteriochlorophyll a were increased significantly. The characteristic absorption maxima of carotenoids from pigment extracts were blueshifted. Iron concentration had little effect on the compositions and relative contents of bacteriochlorophylls a, whereas predominately affected carotenoid compositions, rhodopin was present as major carotenoid component instead of spirillxanthin. Culture tends to accumulate the Cars having shorter conjugated double bonds at the expense of longer conjugated double bonds as the ferric chloride concentration increased. The changes in carotenoid composition were consistent with those of the blue shift of absorption spectra of pigment extracts. CONCLUSION: Rps. palustris CQV97 can produce siderophore and the changes in microbial growth, siderophore production and photosynthetic pigments accumulation of anoxygenic phototrophic bacteria are related to the iron concentration in the medium.

[Quantification of carotenoids of spirilloxanthin series from anoxygenic phototrophic bacteria by substitute reference standard calibration function method].

OBJECTIVE: In this study, we developed a strategy for accurate, rapid and simultaneous quantification of six carotenoids by substitute reference standard. METHODS: We prepared six carotenoid standards of spirilloxanthin series from Rhodopesudomonnas palustris CQV97 by spectrophotometry, thin layer chromatography and HPLC. The simultaneous quantification method for six carotenoids was established by HPLC using tartrazine and lycopene as substitute reference standards. RESULTS: We established the HPLC fingerprinting of carotenoids of spirilloxanthin series. The quantitative calibration function relationships between two substitute reference standards and six carotenoids were explored. Based on the quantitative calibration function relationships, we quantitatively analyzed carotenoid contents of two samples of CQV97 and YL28 strains. The RSD and recovery of carotenoid contents determined by substitute reference standards method were consistent with quantitative analysis of carotenoid standards method. CONCLUSION: The substitute reference standards were capable of accurately transmitting the quantitative relationship of tested samples. The method could realize the simultaneous quantification of six carotenoids.

[Arsenic metabolism in purple nonsulfur bacteria].

OBJECTIVE: To elucidate the arsenic metabolic pathway of purple nonsulfur bacteria (PNSB). METHODS: We investigated the distribution within their genomes, organization, composition, arrangement, core genes and coding proteins of arsenic gene clusters found in complete genome from 17 strains of PNSB by comparing the genomes analysis, and studied the arsenic metabolism in 3 members of PNSB under anaerobic conditions by UV-Vis and HPLC-ICP-MS. RESULTS: Arsenate reduction and arsenite methylation pathways mediated by ars operon are the dominating arsenic metabolic processes. The arsenic gene clusters differ vastly in composition and arrangement. Some members of PNSB evolved two independently families of arsenate reduction genes (arsC). The cells of Rhodopseudomonas palustris CQV97, Rhodobacter azotoformans 134K20 and Rhodobacter capsulatus XJ-1 could reduce As (V) to As (III), whereas As (III) could not be transformed back to As (V). Higher concentration phosphate competitively inhibited arsenate toxicity to cells. CONCLUSION: Our investigations shed light on the evolution and functional implications in arsenic gene clusters of PNSB, and support the notion that arsenate reduction and arsenite methylation appears to be the dominant process in PNSB.

[Fingerprinting analysis of photopigments in purple bacteria].

OBJECTIVE: Photopigments, including carotenoid and bacteriochlorophyll a, are the most important functional units of photosynthesis in purple bacteria. We developed rapid qualitative and quantitative methods to determine photopigments. METHODS: Using Rhodopesudomonas palustris CQV97 as a reference, we used image gray intensity analysis, absorption spectrophotometry, thin layer chromatography (TLC), HPLC and mass spectrometry (MS) for photopigment analysis. RESULTS: The total amount of photopigments increased by 13.5% by using modified acetone-methanol extraction. We developed two types of photopigment fingerprintings by TLC and HPLC, estimated the apparent relative content of each photopigment of fingerprintings, and determined the corresponding relationships between R(f) value of each photopigment on TLC fingerprinting and retention time of each photopigment elution in HPLC fingerprinting. Based on the data from the absorption spectra, MS and related photopigment biosynthetic pathway analysis, we identified 11 photopigments in CQV97 strain. Using this strain as a standard, we analyzed photopigments of the tested samples by TLC or HPLC. It was shown that (1) the relative standard deviation (RSD) of the two methods was less than 5%; (2) the compositions and contents of the theory sample were consistent with that of the standard sample; (3) the photopigment compositions of the real sample was the same as the standard sample, but the photopigment content was different. CONCLUSION: Both of TLC and HPLC analyses for photopigment determination have good stability and repeatability. The fingerprintings analyses are suitable for rapid determination of photopigments of purple bacteria and have important application in control of regulation mechanism for photopigment synthesis.

Anaerobic utilization of phenanthrene by Rhodopseudomonas palustris.

A phenanthrene-utilizing bacterium was anaerobically isolated and identified as Rhodopseudomonas palustris. It tolerated up to 100 mg phenanthrene l(-1) and degraded 50% of 50 mg phenanthrene l(-1) over 10 days. The presence of phenanthrene caused a prolonged lag phase (2-3 days) in cell growth and affected the photopigments biosynthesis, while DMSO (the solvent for phenanthrene) had no impact on cell growth. The cell surface hydrophobicity of the isolate was also increased.

[Identification and characterization of a purple sulfur bacterium from mangrove with rhodopin as predominant carotenoid].

OBJECTIVE: To exploit resources of purple sulfur bacteria in China and further investigate its response mechanism to ecological environment of mangrove. METHODS: Repeated agar shake dilution method, microscope techniques, UV-Vis absorption spectra, thin layer chromatography, HPLC and MS were used. RESULTS: We isolated a purple sulfur bacterium, designated as strain YL28, from a intertidal sediment sample collected from inshore mangrove near Luoyang Bridge of Quanzhou city, Fujian Province of China. Cells were ovoid to rod shaped, 0.5 microm - 1 microm x 2 microm - 3 microm. Color of cell suspensions was reddish-brown. It possessed vesicular intracytoplasmic membrane structures, contained rhodopin and phytylated bacteriochlorophyll a as well as the other two novel bacteriochlorophyll a intermediates. The optimum growth was at 2% - 3.5% NaCl, pH 5.7 - 6.7 and 20 degrees C - 35 degrees C. Photoautotrophically growth anaerobically in the light with sulphide, sulphur, thiosulfate, sulfite as electron donor. Globules of S(0) distributed inside the cells. Photoheterotrophic growth with various organic substrates, especially citrate, glucose, sucrose, fructose and propanol in the presence of sulfide. Nitrogen sources: ammonium salts, N2, urea, glutamate, nitrate and nitrite. Vitamins were not required. Qualitative assessment of IC50 values of chloromycetin, cefazolin, benzene, hydroxy biphenyl, enrofloxacin, acetamiprid, mercuric chloride and cadmium chloride were 70, 100, 20, 20, 3, 170, 5 mg/L and 25 mg/L, respectively. CONCLUSION: Based on phenotype characteristics and 16S rRNA gene sequence similarity of 99% to M. gracile, strain YL28 was identified as novel isolate of M. gracile despite its different physiological characteristics with respect to the species of M. gracile. The organism is possessed of slightly acid tolerance, higher amount of carotenoid of rhodopin and tolerance towards certain antibiotics, pesticide as well as heavy metals.

Synthesis and antimicrobial activities of chitosan/polypropylene carbonate-based nanoparticles.

Antibiotic resistance is an emerging threat to public health. The development of a new generation of antimicrobial compounds is therefore currently required. Here we report a novel antimicrobial polymer of chitosan/polypropylene carbonate nanoparticles (CS/PPC NPs). These were designed and synthesized from readily available chitosan and a reactive oligomer polypropylene carbonate (PPC)-derived epoxy intermediate. By employing a simple and efficient functionalized strategy, a series of micelle-like chitosan-graft-polypropylene carbonate (CS-g-PPC) polymers and chitosan-polypropylene carbonate (CS-PPC) microgels were prepared by reacting mono-/bis-epoxy capped PPC with chitosan. The chemical structure, particle size, and surface charge of the newly synthesized polymers were characterized by infrared (IR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, dynamic light scattering (DLS), and zeta potential measurements. The antimicrobial activities of these nanoparticles were determined in both Gram-positive bacteria (S. aureus) and Gram-negative bacteria (E. coli). Minimum inhibitory concentration (MIC), the nanoparticle concentration needed to completely inhibit the bacterial growth, was found at 128 µg mL-1 to 1024 µg mL-1, strongly depending both on the nature of the epoxy-imine network formed from the functional groups (mono- or bis-capped epoxy groups reacting with amine groups) and the feed ratio of the functional groups (-epoxy/-NH2) between the functionalized PPC and chitosan. No hemolysis was observed at concentrations well in excess of the effective bacteria-inhibiting concentrations. These findings provide a novel strategy to fabricate a new type of nanoantibiotic for antimicrobial applications.

Influences of organic nitrogen on the removal of inorganic nitrogen from complicated marine aquaculture water by Marichromatium gracile YL28.

The sediment-water interface is not only an important location for substrate conversion in a mariculture system, but also a major source of eutrophication. This study aimed to clarify the characteristics of inorganic nitrogen (ammonia, nitrite and nitrate) removal by Marichromatium gracile YL28 in the presence of both organic nitrogen and inorganic nitrogen. The results showed that, in the presence of peptone or urea, seaweed oligosaccharides (SOS) effectively enhanced the ammonia removal capacity of YL28 (6.42 mmol/L) and decreased the residual rate by 54.04% or 8.17%, respectively. With increasing peptone or urea concentrations, the removal of both ammonia and nitrate was gradually inhibited, and the residual rates of ammonia and nitrate reached 22.56-34.36% and 12.03-15.64% in the peptone system and 20.65-24.03% and 12.20-13.21% in the urea system, respectively. However, in the control group the residual rates of ammonia and nitrate reached 11.97% and 5.12%, respectively. In addition, the concentrations of peptone and urea did not affect nitrite removal, and YL28 displayed better cell growth and nitrogen removal activity in the presence of bait and SOS. Overall, the ability of YL28 to remove inorganic nitrogen was enhanced in the presence of organic nitrogen.

Effects of Supplement of Marichromatium gracile YL28 on Water Quality and Microbial Structures in Shrimp Mariculture Ecosystems.

The elevated NH3-N and NO2-N pollution problems in mariculture have raised concerns because they pose threats to animal health and coastal and offshore environments. Supplement of Marichromatium gracile YL28 (YL28) into polluted shrimp rearing water and sediment significantly decreased ammonia and nitrite concentrations, showing that YL28 functioned as a novel safe marine probiotic in the shrimp culture industry. The diversity of aquatic bacteria in the shrimp mariculture ecosystems was studied by sequencing the V4 region of 16S rRNA genes, with respect to additions of YL28 at the low and high concentrations. It was revealed by 16S rRNA sequencing analysis that Proteobacteria, Planctomycete and Bacteroidetes dominated the community (>80% of operational taxonomic units (OTUs)). Up to 41.6% of the predominant bacterial members were placed in the classes Gammaproteobacteria (14%), Deltaproteobacteria (14%), Planctomycetacia (8%) and Alphaproteobacteria (5.6%) while 40% of OTUs belonged to unclassified ones or others, indicating that the considerable bacterial populations were novel in our shrimp mariculture. Bacterial communities were similar between YL28 supplements and control groups (without addition of YL28) revealed by the ß-diversity using PCoA, demonstrating that the additions of YL28 did not disturb the microbiota in shrimp mariculture ecosystems. Instead, the addition of YL28 increased the relative abundance of ammonia-oxidizing and denitrifying bacteria. The quantitative PCR analysis further showed that key genes including nifH and amoA involved in nitrification and nitrate or nitrite reduction significantly increased with YL28 supplementation (p < 0.05). The supplement of YL28 decreased the relative abundance of potential pathogen Vibrio. Together, our studies showed that supplement of YL28 improved the water quality by increasing the relative abundance of ammonia-oxidizing and denitrifying bacteria while the microbial community structure persisted in shrimp mariculture ecosystems.

ArsM-mediated arsenite volatilization is limited by efflux catalyzed by As efflux transporters.

Arsenic (As) methylation is regarded as an efficient strategy for As contamination remediation by As volatilization. However, most microorganisms display low As volatilization efficiency, which is possibly linked to As efflux transporters competing for cytoplasmic As(III) as a substrate. Here, we developed two types of As biosensors in Escherichia coli to compare the As efflux rate of three efflux transporters and to further investigate the correlation between As efflux rates and As volatilization. The engineered As-sensitive E. coli AW3110 expressing arsBRP, acr3RP or arsBEC displayed a higher As resistance compared to the control. The fluorescence intensity was in a linear correlation in the range of 0-2.0 µmol/L of As(III). The intracellular As(III) concentration was negatively related to As efflux activity of As efflux transporter, which was consistent with the As resistance assays. Moreover, arsM derived from R. palustris CGA009 was subsequently introduced to construct an E. coli AW3110 co-expressing arsB/acr3 and arsM, which exhibited higher As(III) resistance, lower fluorescence intensity and intracellular As concentration compared to the engineered E. coli AW3110 expressing only arsB/acr3. The As volatilization efficiency was negatively related to As efflux activity of efflux transporters, the recombinants without arsB/acr3 displayed the highest rate of As volatilization. This study provided new insights into parameters affecting As volatilization with As efflux being the main limiting factor for As methylation and subsequent volatilization in many microorganisms.

[Regulation mechanism of photopigments biosynthesis via light and oxygen in Rhodobacter azotoformans 134K20].

OBJECTIVE: To provided a reliable and sensitive method and a series of specific absorption spectra data of different carotenoids and bacteriochlorophylla in anxoygenic phototrophic bacteria, and reveal mechanism of regulation of photosynthetic pigments by light and oxygen in Rhodobacter azotoformans 134K20. METHODS: The metabolic diversity of photopigments regulated by oxygen and light was investigated by means of UV-VIS spectra and thin layer chromatography. RESULTS: The highest cell yield of strain 134K20 was obtained under aerobic conditions in the light. Nine types of photopigments including three yellow pigments, one red pigment, one purple pigment, two green pigments, and two blue pigments were synthesized, and yellow pigments synthetic genes were expressed on the higher level anaerobically in the light. Under aerobic conditions, the synthetic genes of two new red and one new purple pigment were triggered and expressed on the higher level, but the biosynthesis of yellow, blue and green pigments were inhibited by oxygen, another nine pigments including two yellow pigments, three red pigments, two purple pigments, one green pigment, and one blue pigment were synthesized. One yellow pigment was only produced in dark aerobic culture, the other pigments were the same as in dark aerobic cultures. CONCLUSION: PpsR photopigment suppression regulation system regulated expression of photosynthetic genes via light and oxygen in Rhodobacter azotoformans 134K20. The yellow and red pigments belong to carotenoid series. The first yellow pigment belongs to spheroidene series. The other two yellow pigments are new carotenoids. Yellow pigments are capable of dissolving in different organic solvents. Red pigments belong to new spheroidenone series. The three red pigments are different in polarity, peak shape and peak value. Fine structures of red pigments only are appeared in hexane. The purple pigmments with polarity are identified as bacteriopheophytins. The blue and green pigments are four kinds of bacteriochlorophyll a intermediates. Diethylether and methanol is suitable for carotenoids extraction. The identification of bacteriochlorophyll a intermediates can be easily performed by polarity analysis.

[Progress on pigment-protein complexes from anoxygenic phototrophic bacteria--a review].

Anoxygenic phototrophic bacteria have evolved highly efficient systems-membrane-located pigment-protein complexes which can convert sunlight into chemical energy that they can use and also benefit other organisms. More and more attentions have been paid to pigment-protein complexes of anoxygenic phototrophic bacteria in recent years. We summarized the current opinions in the pigment-protein complexes from anoxygenic phototrophic bacteria, including their chemical compositions, crystal structures and functions, homology of protein sequence. In particular, we depicted the novel light-harvesting complexes namely LH3 and LH4. The problems and prospects about the pigment-protein complexes have also been addressed in this review.