• Economic and Environmental Geology
• /
• v.47 no.4
• /
• pp.431-439
• /
• 2014

The purposes of this research were to investigate the enrichment of metal-reducing bacteria from KURT groundwater and the identification of the microbial diversity by 16S rRNA as well as to examine microbial Fe(III)/Mn(IV) reduction and to analyze morphological features of interactions between microbes and precipitates and their mineralogical composition. To cultivate metal-reducing bacteria from groundwater sampled at the KURT in S. Korea, different electron donors such as glucose, acetate, lactate, formate, pyruvate and Fe(III)-citrate as an electron accepter were added into growth media. The enriched culture was identified by 16S rRNA gene sequence analysis for the diversity of microbial species. The effect of electron donors (i.e., glucose, acetate, lactate, formate, pyruvate) and electron acceptors (i.e., akaganeite, manganese oxide) on microbial iron/manganese reduction and biomineralization were examined using the 1st enriched culture, respectively. SEM, EDX, and XRD analyses were used to determine morphological features, chemical composition of microbes and mineralogical characteristics of the iron and manganese minerals. Based on 16S rRNA gene analysis, the four species, Fusibacter, Desulfuromonas, Actinobacteria, Pseudomonas sp., from KURT groundwater were identified as anaerobic metal reducers and these microbes precipitated metals outside of cells in common. XRD and EDX analyses showed that Fe(III)-containing mineral, akaganeite (${\beta}$-FeOOH), reduced into Fe(II)/Fe(III)-containing magnetite ($Fe_3O_4$) and Mn(IV)-containing manganese oxide (${\lambda}-MnO_2$) into Mn(II)-containing rhodochrosite ($MnCO_3$) by the microbes. These results implicate that microbial metabolism and respiratory activities under anaerobic condition result in reduction and biomineralization of iron and manganese minerals. Therefore, the microbes cultivated from groundwater in KURT might play a major role to reduce various metals from highly toxic, mobile to less toxic, immobile.

• Journal of the Mineralogical Society of Korea
• /
• v.15 no.3
• /
• pp.231-240
• /
• 2002

Microbial metal reduction influences the biogeochemical cycles of carbon and metals as well as plays an important role in the bioremediation of metals, radionuclides, and organic contaminants. The use of bacteria to facilitate the production of magnetite nanoparticles and the formation of carbonate minerals may provide new biotechnological processes for material synthesis and carbon sequestration. Metal-reducing bacteria were isolated from a variety of extreme environments, such as deep terrestrial subsurface, deep marine sediments, water near Hydrothemal vents, and alkaline ponds. Metal-reducing bacteria isolated from diverse extreme environments were able to reduce Fe(III), Mn(IV), Cr(VI), Co(III), and U(VI) using short chain fatty acids and/or hydrogen as the electron donors. These bacteria exhibited diverse mineral precipitation capabilities including the formation of magnetite ($Fe_3$$O_4$), siderite ($FeCO_3$), calcite ($CaCO_3$), rhodochrosite ($MnCO_3$), vivianite [$Fe_3$($PO_4$)$_2$ .$8H_2$O], and uraninite ($UO_2$). Geochemical and environmental factors such as atmospheres, chemical milieu, and species of bacteria affected the extent of Fe(III)-reduction as well as the mineralogy and morphology of the crystalline iron mineral phases. Thermophilic bacteria use amorphous Fe(III)-oxyhydroxide plus metals (Co, Cr, Ni) as an electron acceptor and organic carbon as an electron donor to synthesize metal-substituted magnetite. Metal reducing bacteria were capable of $CO_2$conversion Into sparingly soluble carbonate minerals, such as siderite and calcite using amorphous Fe(III)-oxyhydroxide or metal-rich fly ash. These results indicate that microbial Fe(III)-reduction may not only play important roles in iron and carbon biogeochemistry in natural environments, but also be potentially useful f3r the synthesis of submicron-sized ferromagnetic materials.

• Applied Biological Chemistry
• /
• v.48 no.3
• /
• pp.189-200
• /
• 2005

Biological nitrogen fixation is an important process for academic and industrial aspects. This review will briefly compare industrial and biological nitrogen fixation and cover the characteristics of biological nitrogen fixation studied in Azotobacter vinelandii. Various organisms can carry out biological nitrogen fixation and recently the researches on the reaction mechanism were concentrated on the free-living microorganism, A. vinelandii. Nitrogen fixation, which transforms atmospheric $N_2$ into ammonia, is chemically a reduction reaction requiring electron donation. Nitrogenase, the biological nitrgen fixer, accepts electrons from biological electron donors, and transfers them to the active site, FeMo-cofactor, through $Fe_4S_4$ cluster in Fe protein and P-cluster in MoFe protein. The electron transport and the proton transport are very important processes in the nitrogenase catalysis to understand its reaction mechanism, and the interactions between FeMo-cofactor and nitrogen molecule are at the center of biological nitrogen fixation mechanism. Spectroscopic studies including protein X-ray crystallography, EPR and $M{\ddot{o}}ssbauer$, biochemical approaches including substrate and inhibitor interactions as well as site-directed mutation study, and chemical approach to synthesize the FeMo-cofactor model compounds were used for biological nitrogen fixation study. Recent research results from these area were presented, and finally, a new nitrogenase reaction mechanism will be proposed based on the various research results.

• Korean Journal of Microbiology
• /
• v.48 no.1
• /
• pp.42-47
• /
• 2012

Hydrothermal vents (HTV) provide special environments for evolution of lives independent on solar energy. HTV samples were gained from Tofua arc trench in Tonga, South Pacific. We investigated archaeal diversity enriched using combinations of various electron donors (yeast extract and $H_2$) and electron acceptors [Iron (III), elemental sulfur ($S^0$) and nitrate. PCR amplification was performed to detect archaeal 16S rRNA genes after the cultures were incubated $65^{\circ}C$ and $80^{\circ}C$ for 2 weeks. The cultures showing archaeal growth were transferred using the dilution-to-extinction method. 16S rRNA gene PCR-Denaturing Gradient Gel Electrophoresis was used to identify the enriched archaea in the highest dilutions where archaeal growth was observed. Most of cultured archaea belonged to genus of Thermococcus (T. alcaliphilius, T. litoralis, T. celer, T. barossii, T. thoreducens, T. coalescens) with 98-99% 16S rRNA gene similarities. Interestingly, archaeal growth was observed in the cultures with Iron (III) and nitrate as an electron acceptor. It was supposed that archaea might use the elemental sulfur generated from oxidation of the reducing agent, sulfide. To cultivate diverse archaea excluding Thermococcus, it would be required to use other reducing agents instead of sulfide.

• Korean Journal of Microbiology
• /
• v.50 no.3
• /
• pp.254-260
• /
• 2014

Sulfur compound includes major electron acceptors for anaerobic respiration. In this study, cultivation-based study on sulfate- and sulfur-reducing bacteria of various wetlands of Korea was attempted. To isolate sulfate- and sulfur-reducing bacteria, anaerobic roll tube method was used to obtain typical black colonies of sulfate- and sulfur-reducing bacteria. Total 11 strains obtained were tentatively identified based on comparative 16S rDNA similarity and physiological property analysis. All sulfate-reducing bacteria (8 strains) belonged to genus Desulfovibrio with >99% 16S rDNA similarities. Three sulfur reducing bacteria were also isolated: two and one isolates were affiliated with Sulfurospirillum and Desulfitobacterium, respectively. These sulfate- and sulfur-reducing bacteria were able to utilize lactate and pyruvate and sulfite and thiosulfate as common electron donors and electron acceptors, respectively. This case study will provide fundamental information for obtaining useful indigenous sulfate- and sulfur-reducing bacteria from Korean wetlands employing various combinations of cultivation conditions.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.5
• /
• pp.580-586
• /
• 2000

A soluble Cr(VI) reductase was purified from the Cr(VI) reducing strain Escherichia coli ATCC33456 by ammonium sulfate fractionation, and chromatographies on Q-Sepharose FF, Cibacron blue 3GA dye affinity, Mono-Q 5/5, and Superdex 200 HR 10/30 columns. The estimated molecular mass of the purified enzyme was 27 kDa on SDS-polyacrylamide gel electrophoresis and 54 kDa on gel filtration, thus indicating a dimeric structure. The isoelectric point of the enzyme was pH 4.85. The optimum reaction pH and storage pH were both 7.0, the optimum reaction temperature was $37^{\circ}C$, and the storage temperature was $4^{\circ}C$. NADH and NADPH both served as electron donors for the reductase, with $V_{max}$ of 68.3 ${\mu}M$ Cr(VI)/min/mg protein and Km of 7.6 $\mu$M using HADH, and Vmax of 42.3 ${\mu}M$ Cr(VI)/min/mg protein and Km of 14.6 $\muM$ using NADPH. When 1 mM EDTA was added, the Cr(VI) reducing activity increased 4-fold.

• Bulletin of the Korean Chemical Society
• /
• v.32 no.7
• /
• pp.2279-2285
• /
• 2011

The geometries, electronic structures and absorption spectra of the two organic triphenylamine-based dyes TA-St-CA and TA-DM-CA, containing identical electron donors and acceptors but the different conjugated bridges, were studied by density functional theory (DFT) at the B3LYP and PBE1PBE levels, respectively. The influence of para-orientating methoxyl units on the electronic structures and light absorption properties of the dyes and the consequent photovoltaic performance of the dye-sensitized solar cells (DSSCs) were investigated in detail. The results indicate that the introduction of the para-orientating methoxyl units into the conjugated bridge induces the increased absorption wavelength as well as the more negative EHOMO corresponding to the bigger driving force $(E_{I^-/I^-_3}-E_{HOMO})$ for dye reduction, which together improve the photovoltaic performance of TA-DM-CA, although there is a decline of the open circuit voltage caused by the more negative $E_{LUMO}$.

• Journal of Korean Society on Water Environment
• /
• v.21 no.6
• /
• pp.669-675
• /
• 2005

Biological nitrate removal from groundwater was investigated in the biofilters packed with both gravel/sand and plastic media. Removal of particles and turbidity were also investigated in the 2-stage biofilter system consisted of biofilter and subsequent sand filter. In the single biofilter packed with gravel and sand, nitrate removal efficiency was dropped with the increase of filtration velocity and furthermore, nitrite concentration increased up to 3.2 mg-N/L at 60 m/day. Denitrification rate at the bottom layer below 25 cm was faster 8 times than upper layer in the up-flow biofilter. Nitrite build-up, due to the deficiency of organic electron donors, occurred at the upper layer of bed. Besides DO concentration and organic carbon, contact time in media was the main factor for nitrate removal in a biofilter. The most of the effluent particles from biofilter was in the range from 0.5 to $2.0{\mu}m$, which resulted in high turbidity of 1.8 NTU. However, sand filter followed by biofilter efficiently performed the removal of particles and turbidity, which could reduce the turbidity of final filtrate below 0.5 NTU. Influent nitrate was removed completely in the 2-stage biofilter and no nitrite was detected.

• Microbiology and Biotechnology Letters
• /
• v.11 no.3
• /
• pp.211-216
• /
• 1983

Rhodopseudomonas sphaeroides K-7 evolves large quantities of molecular hydrogen under anaerobic and light illuminated conditions in the presence of utilizable organic compounds as electron donors. Photoevolution of molecular hydrogen was strictly dependent on light as the activity of nitrogenase in this organism. Both of these were inhibited to the nearly same extent at varying concentrations of ammonium ion which also depressed nitrogenase synthesis. In the reaction mixtures devoid of molybdenum ion which is known as the component of nitrogenase, hydrogen evolution also decreased similarly like nitrogenase activity. Photoevolution of molecular hydrogen appeared to have no relationship with hydrogenase activity and bacteriocholophyll content and it was markedly inhibited under the atmosphere of $C_2$H$_2$, $N_2$ or $O_2$. The results strongly indicate that hydrogen evolution by R. sphaeroides K-7 might be catalyzed by nitrogenase. Both hydrogen evolution and nitrogenase activity were largely influenced by the nutritional history of the resting cells. From which we propose that glutamate might play an important role in the regulation of nigrogenase activity in vivo.

• Journal of the Korea Organic Resources Recycling Association
• /
• v.16 no.3
• /
• pp.75-82
• /
• 2008

Two UASB reactors were operated to investigate the effect of high concentration of sulfate on anaerobic digestion of propionate using an upflow anaerobic sludge blanket (UASB) reactor. An organic loading rate of $1.2kg\;COD/m^3{\cdot}d$ and a hydraulic retention time of 1.6 d were maintained during this study. In the absence of sulfate, the UASB reactor achieved about 95% removal of chemical oxygen demand whereas in the presence of $2,000\;SO_4^{2-}mg/L$, the COD removal rate decreased to 83% due probably to the inhibition of dissolved sulfide inhibition. Interactions between the methane producing bacteria (MPB) and sulfate reducing bacteria (SRB) were measured to investigate the competition between MPB and SRB. The MPB consumed average 58% of the available electron donors at $COD/SO_4^{2-}$ ratio of 1. Propionate was consumed mainly by SRB, converting sulfate into sulfide and suppressing the methane production. The specific methanogenic activity (SMA) using acetate and propionate increased as microorganism acclimated to the substrate.