Light-driven nitrous oxide production via autotrophic denitrification by self-photosensitized Thiobacillus denitrificans.

N2O (Nitrous oxide, a booster oxidant in rockets) has attracted increasing interest as a means of enhancing energy production, and it can be produced by nitrate (NO3-) reduction in NO3--loading wastewater. However, conventional denitrification processes are often limited by the lack of bioavailable electron donors. In this study, we innovatively propose a self-photosensitized nonphototrophic Thiobacillus denitrificans (T. denitrificans-CdS) that is capable of NO3- reduction and N2O production driven by light. The system converted >72.1 ±â€¯1.1% of the NO3--N input to N2ON, and the ratio of N2O-N in gaseous products was >96.4 ±â€¯0.4%. The relative transcript abundance of the genes encoding the denitrifying proteins in T. denitrificans-CdS after irradiation was significantly upregulated. The photoexcited electrons acted as the dominant electron sources for NO3- reduction by T. denitrificans-CdS. This study provides the first proof of concept for sustainable and low-cost autotrophic denitrification to generate N2O driven by light. The findings also have strong implications for sustainable environmental management because the sunlight-triggered denitrification reaction driven by nonphototrophic microorganisms may widely occur in nature, particularly in a semiconductive mineral-enriched aqueous environment.

Direct current stimulation of Thiobacillus ferrooxidans bacterial metabolism in a bioelectrical reactor without cation-specific membrane.

A bioelectrical reactor without cation-specific membrane was designed to test effects of direct electrical current on growth of Thiobacillus ferrooxidans bacterium. The results indicated that the cell significantly enhanced the growth of T. ferrooxidans. At a current of 30 mA, the maximum cells density reached 1.39 x 10(9)cells/mL within 84 h, which was 10 times faster than under a conventional cultivation method, in which electrical current is not used. A lag phase during the growth of T. ferrooxidans was observed when direct electrical current was applied, and the lag phase became longer under higher current intensity.

Enhanced UV sensitivity of Thiobacillus ferrooxidans resulting from caffeine and acriflavine treatment of irradiated cells.

This study was aimed at identifying the roles of caffeine and acriflavine, two repair inhibitors, on UV sensitivity of iron-oxidizing Thiobacillus ferrooxidans ATCC 13728. The UV-dose response survival curve was inflected in nature, suggesting the population heterogeneity of the isolate. Caffeine and acriflavine potentiated the UV-induced killing of the organism. With the increase in concentrations of these compounds, the extent of survival decreased. Similarly, the inhibitory effects of caffeine and acriflavine increased with the increase in dose of UV-irradiation. The cells irradiated with 10 s (equivalent to 5.6 x 10(-5) J/m2/s) of UV-exposure tended to become resistant to the inhibitory effects of caffeine and acriflavine, as evidenced by the time course study of recovery. The cells appear to stage a dramatic recovery from UV damage in the presence of caffeine (3.0 mg/ml) and acriflavine (20 microg/ml) over a period of 25-30 h and 35-40 h respectively, when grown in the presence of energy sources.

Construction and characterization of a recA mutant of Thiobacillus ferrooxidans by marker exchange mutagenesis.

To construct Thiobacillus ferrooxidans mutants by marker exchange mutagenesis, a genetic transfer system is required. The transfer of broad-host-range plasmids belonging to the incompatibility groups IncQ (pKT240 and pJRD215), IncP (pJB3Km1), and IncW (pUFR034) from Escherichia coli to two private T. ferrooxidans strains (BRGM1 and Tf-49) and to two collection strains (ATCC 33020 and ATCC 19859) by conjugation was analyzed. To knock out the T. ferrooxidans recA gene, a mobilizable suicide plasmid carrying the ATCC 33020 recA gene disrupted by a kanamycin resistance gene was transferred from E. coli to T. ferrooxidans ATCC 33020 by conjugation under the best conditions determined. The two kanamycin-resistant clones, which have retained the kanamycin-resistant phenotype after growth for several generations in nonselective medium, were shown to have the kanamycin resistance gene inserted within the recA gene, indicating that the recA::Omega-Km mutated allele was transferred from the suicide plasmid to the chromosome by homologous recombination. These mutants exhibited a slightly reduced growth rate and an increased sensitivity to UV and gamma irradiation compared to the wild-type strain. However, the T. ferrooxidans recA mutants are less sensitive to these physical DNA-damaging agents than the recA mutants described in other bacterial species, suggesting that RecA plays a minor role in DNA repair in T. ferrooxidans.

Modification of radiosensitivity of chlorpromazine with biological metal ions in Thiobacillus ferrooxidans.

Effect of chlorpromazine with biological metal ions, viz. calcium, magnesium, zink and copper was studied on T. ferrooxidans cell system. Chlorpromazine, calcium and magnesium alone could produce radioprotection. Maximum radioprotection was exhibited by chlorpromazine at lower concentration while copper and zink offered radiosensitization. However, combination of chlorpromazine with all biological metal ions exhibited radiosensitization. Dose modifying factor by chlorpromazine at lower concentration (0.025 mM) was 0.754 while in combination with Ca2+, Mg2+, Cu2+ and Zn2+ was 1.08, 1.25, 1.37 and 1.389 respectively. The possible interaction between chlorpromazine and biological metal ions is discussed at cellular membrane level.

Effects of 60Co gamma-rays, ultraviolet light, and mitomycin C on Halobacterium salinarium and Thiobacillus intermedius.

Lethal effects of 60Co gamma-rays, UV light, and mitomycin C on two kinds of bacteria, Halobacterium salinarium which grows in highly concentrated salt media and Thiobacillus intermedius which requires reduced sulfur compounds, were studied and compared with those on Escherichia coli B/r. D37 values for H. salinarium, T. intermedius and E. coli B/r were 393, 150, and 92 Gy, respectively, by exposure to 60Co gamma-rays. They were 212, 38, and 10 J/m2, respectively, by exposure to UV light and 2.36, 0.25, and 0.53 microgram/ml/h, respectively, by exposure to mitomycin C. Against these agents, H. salinarium was much more resistant than T. intermedius and E. coli B/r.

Inhibition of oxygen utilization and destruction of ubiquinone by ultraviolet irradiation of Thiobacillus thiooxidans.

Ultraviolet (UV) irradiation inhibited sulfur oxidation by cells of Thiobacillus thiooxidans. Sulfur-oxidizing activity decreased as the exposure time to UV light increased. A loss of the ability of cells of fix CO(2) paralleled the loss of sulfur-oxidizing activity. UV light photoinactivated ubiquinone purified from T. thiooxidans. The same percentage of sulfur-oxidizing activity and ubiquinone was destroyed after 15 min of UV exposure. Both the photoinactivation of sulfur oxidation and ubiquinone followed first-order reaction kinetics. The specific rate constants for both photoinactivations were nearly equal. Cells completely inactivated by UV light contained no ubiquinone. Ubiquinone was found to be a component of the cell wall-membrane complex.