• Journal of Microbiology and Biotechnology
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• v.7 no.6
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• pp.391-396
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• 1997

Two series of carbon sources, linear primary $C_1$~$C_9$ alcohols and linear $C_2$~$C_{10}$ monocarboxylic acids were tested for PHA synthesis in Paracoccus denitrificans. The results showed that the growth-associated synthesis of PHA could be referred only to the carbon sources with odd number of carbon except methanol. For all carbon sources with even number of carbon, nitrogen limitation was required to induce PHA synthesis in P. denitrificans. Poly(3-hydroxyvalerate)[P(3HV)] homopolymer was synthesized from $C_5$, $C_7$, and $C_9$ while growing in the presence of nitrogen, but the nitrogen depletion in the later growth period incorporated 3-hydroxybutyrate(3HB) unit into the polymer chain. The optimum C/N ratio for P(3HV) homopolymer production was found to be 10 when the strain was grown on 10 ml/l of valeric acid for 96 h. P. denitrificans synthesized P(3HB-co-3HV) copolymers from n-hexanoic and n-octanoic acid. The microstructural characterics of the P(3HB-co-3HV) copolymer from n-propanol was investigated using $^13C$-nuclear magnetic resonance spectroscopy, showing a structural heterogeneity.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.46-51
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• 2010

Coenzyme Q10 (CoQ10) is an essential lipid-soluble component of membrane-bound electron transport chains. CoQ10 is involved in several aspects of cellular metabolism and is increasingly being used in therapeutic applications for several diseases. Despite the recent accomplishments in metabolic engineering of Escherichia coli for CoQ10 production, the production levels are not yet competitive with those by fermentation or isolation. So we tested several microorganisms obtained from the KCTC of Biological Resource Center to find novel sources of strain-development for CoQ10-production. Then we selected two strains, Paracoccus denitrificans (KCTC 2530) and Asaia siamensis (KCTC 12914), and tested to optimize the CoQ10 production conditions. Among the carbon sources tested, CoQ10 production was the highest when fructose was supplied about 4% concentration. Yeast extract produced the highest CoQ10 production about 2% concentration. The highest CoQ10 production was obtained at pH 6.0 for P. denitrificans and pH 8.0 for A. siamensis. And two strains showed the highest CoQ10 production at $30^{\circ}C$, but the highest DCW was obtained at $37^{\circ}C$. In the fed-batch culture, P. denitrificans yielded $14.34{\pm}0.473$ mg and A. siamensis yielded $12.53{\pm}0.231$ mg of final CoQ10 production.

• KSBB Journal
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• v.16 no.3
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• pp.290-294
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• 2001

The removal of nitrogen compounds from waste water is essential and is often accomplished by biological process. The denitrifying bacterium, Paracoccus denitrificans (KCTC 2530), was employed to study the characteristics and the denitrification differences of Permeabilized strains and untreated strains. The permeabilization rate increased with increasing toluene concentration, but some part of the toluene contributed to denaturing the datachment of proteins from the plasma membrane. Permeabilized Paracoccus denitrificans had long lag phase and high specific growth rate in cultivation, and showed excellent denitrification characteristic compared with untreated strains. But, in both cases, the denitrification ability was significantly reduced after 4 or 5 denitrifications. It seems that the strains fall into the death phase when the nutrient was exhausted. When the nutrient recovered to its initial level, the denitrification ability also recovered to the normal level. The results obtained were encouraging enough to apply to practical water treatment situation.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.16 no.2
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• pp.68-74
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• 1983

A moderately halophilic bacterium, Listeria denitrificans HB-38, isolated from mud on the seashore in Sooyoung bay, Pusan, showed the requirement of $4\%$ sodium chloride for cell growth in a medium with salts typical of a marine environment, and showed that of $10\%$ in a medium with salts typical of a terrestrial environment. The optimum temperature and pH for growth were $40^{circ}C$ and pH 7.5 in the medium containing $10\%$ sodium chloride and ions typical of a terrestrial environment. Sodium chloride as a protoplast stabilizer gave more stability than sorbitol or sucrose, meanwhile the protoplast did not require higher concentration of stabilizer than that of E. coli protoplast. Succinic dehydrogenase of HB-38 had a halophilic property showing maximal activity in the presence of $9\%$ sodium chloride. The strain HB-38 did not harbor an extrach-romosomal DNA.

• Korean Journal of Food Science and Technology
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• v.30 no.4
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• pp.902-907
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• 1998

In this study the anaerobic degradation of nitrate by in GFM (gel and foam matrix) and bead gel immobilized Paracoccus denitrificans DSM 65 in continous culture was conducted. A novel GFM immobilization system was developed in order to improve conventional system (bead). With increasing nitrate concentration in water, the nitrate reduction rate was increased. The observed maximum denitrification rate by in GFM immobilized cells was 177 mg/L h in buffered water, while that was 33 mg/L h in tap water. In comparison with bead system the reduction activity by GFM system showed $1.2{\sim}2.1$ times better. The denitrification activity was not changed after 16 days storage at $5^{\circ}C$ and also showed better activity than that of free cells or even bead immobilized cells.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2000.11a
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• pp.209-212
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• 2000

As a bench-scale study, transformation of nitrate to nitrogen gas under anoxic condition was determined by using autotrophic denitrifiers containing Thiobacillus denitrificans and elemental sulfur as an electron donor. The research objective is to measure the basic kinetic parameters of autotrophic denitrification reaction on the removal efficiency of nitrate. The results showed that nitrate was almost completely transformed to nitrite in the first 4 days of column operation. After 2 days of accumulation of nitrite, its concentration slowly decreased and the compound was detected less than 0.5 mg/L in 14 days. In the experiment, sulfate concentration in the effluent was the 70~90 mg-S/L and the pH was maintained around pH 7.5. When nitrate concentration of bank filtrate in the real field is considered, this sulfate concentration seems to be acceptable. At 17 cm from the bottom of the column, the effluent showed the highest nitrite concentration, and nitrate concentration decreased rapidly to the Point of 33 cm from the bottom. The results suggest that an appropriate thickness of permeable reactive barriers is about 30 cm.

• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1154-1162
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• 2021

The transcriptional capacities of target genes are strongly influenced by promoters, whereas few studies have focused on the development of robust, high-performance and cross-species promoters for wide application in different bacteria. In this work, four novel promoters (P_k.rtufB, P_k.r1, P_k.r2, and P_k.r3) were predicted from Ketogulonicigenium robustum and their inconsistency in the -10 and -35 region nucleotide sequences indicated they were different promoters. Their activities were evaluated by using green fluorescent protein (gfp) as a reporter in different species of bacteria, including K. vulgare SPU B805, Pseudomonas putida KT2440, Paracoccus denitrificans PD1222, Bacillus licheniformis and Raoultella ornithinolytica, due to their importance in metabolic engineering. Our results showed that the four promoters had different activities, with P_k.r1 showing the strongest activity in almost all of the experimental bacteria. By comparison with the commonly used promoters of E. coli (tufB, lac, lacUV5), K. vulgare (Psdh, Psndh) and P. putida KT2440 (JE111411), the four promoters showed significant differences due to only 12.62% nucleotide similarities, and relatively higher ability in regulating target gene expression. Further validation experiments confirmed their ability in initiating the target minCD cassette because of the shape changes under the promoter regulation. The overexpression of sorbose dehydrogenase and cytochrome c551 by P_k.r1 and P_k.r2 resulted in a 22.75% enhancement of 2-KGA yield, indicating their potential for practical application in metabolic engineering. This study demonstrates an example of applying bioinformatics to find new biological components for gene operation and provides four novel promoters with broad suitability, which enriches the usable range of promoters to realize accurate regulation in different genetic backgrounds.

• Korean Journal of Soil Science and Fertilizer
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• v.39 no.6
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• pp.409-414
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• 2006

Sulfur-oxidizing bacteria were enumerated and isolated from a steady-state anaerobic master culture reactor (MCR) operated for over six months under a semi-continuous mode and nitrate-limiting conditions using thiosulfate as an electron donor. Most are Gram-negative bacteria, which have sizes up to 12 m. Strains AD1 and AD2 were isolated from the plate count agar (PCA), and strains BD1 and BD2 from the solid thiosulfate/nitrate medium. Based on the morphological, physiological, FAME and 16S rDNA sequence analyses, the two dominant strains, AD1 and AD2, were identified as Paracoccus denitrificans and Paracoccus versutus (formerly Thiobacillus versutus), respectively. From the physiological results, glucose was assimilated by both strains AD1 and AD2. Heterotrophic growth of strains AD1 and AD2 could be a more efficient way of obtaining a greater amount of biomass for use as an inoculum. Even though facultative autotrophic bacteria grow under heterotrophic conditions, autotrophic denitrification would not be reduced.

• Proceedings of the Korea Water Resources Association Conference
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• 2007.05a
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• pp.46-52
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• 2007

As flue gas desulfurization (FGD) wastewater contains high concentrations of nitrate and is very low in organic carbon, the feasibility of nitrate removal by autotrophic denitrification using Thiobacillus denitrificans was studied. This autotrophic bacteria oxidizes elemental sulfur to sulfate while reducing nitrate to elemental nitrogen gas, thereby eliminating the need for addition of organic compounds such as methanol. Owing to the unusually high concentrations of dissolved salts $(Ca^{2+},\;Mg^{2+},\;Na^+,\;K^+,\;B^+,\;SO_4^{2-},\;Cl^-,\;F^-,)$ in the FGD wastewater, extensive laboratory-scale and pilot-scale tests were carried out in sulfur-limestone reactors (1) to determine the effect of salinity on autotrophic denitrification, (2) to evaluate the use of limestone for pH control and as source of inorganic carbon for microbial growth, and, (3) to find the optimum environmental and operational conditions for autotrophic denitrification of FGD wastewater. The experimental results demonstrated that (1) autotrophic denitrification is not inhibited up to 1.8 mol total dissolved salt content; (2) inorganic carbon and inorganic phosphorus must be present in sufficiently high concentrations; (3) limestone can supply effective buffering capacity and inorganic carbon; (4) the high calcium concentration may interfere with pH control, phosphorus solubility and limestone dissolution, hence requiring pretreatment of the FGD wastewater; and, 5) under optimum conditions, complete autotrophic denitrification of FGD wastewater was obtained in a sulfur-limestone packed bed reactor with a sulfur:limestone volume ratio of 2:1 for volumetric loading rates up to 400g $NO_{3^-}N/m^3.d$. The interesting interactions between autotrophic denitrification, pH, alkalinity, and the unusually high calcium and boron content of the FGD wastewater are highlighted. The engineering significance of the results is discussed.

• Journal of Soil and Groundwater Environment
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• v.14 no.3
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• pp.40-46
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• 2009

The applicability of a well-type autotrophic sulfur-oxidizing reactive barrier (L $\times$ W $\times$ D = $3m\;{\times}\;4\;m\;{\times}\;2\;m$) as a long-term treatment option for nitrate removal in groundwater was evaluated. Pilot-scale (L $\times$ W $\times$ D = $8m\;{\times}\;4\;m\;{\times}\;2\;m$) flow-tank experiments were conducted to examine remedial efficacy of the well-type reactive barrier. A total of 80 kg sulfur granules as an electron donor and Thiobacillus denitrificans as an active bacterial species were prepared. Thiobacillus denitrificans was successfully colonized on the surface of the sulfur granules and the microflora transformed nitrate with removal efficiency of ~12% (0.07 mM) for 11 days, ~24% (1.3 mM) for 18 days, ~45% (2.4 mM) for 32 days, and ~52% (2.8 mM) for 60 days. Sulfur granules attached to Thiobacillus denitrificans were used to construct the well-type reactive barrier comprising three discrete barriers installed at 1-m interval downstream. Average initial nitrate concentrations were 181 mg/L for the first 28 days and 281 mg/L for the next 14 days. For the 181 mg/L (2.9 mM) plume, nitrate concentrations decreased by ~2% (0.06 mM), ~9% (0.27 mM), and ~15% (0.44 mM) after $1^{st}$, $2^{nd}$, and $3^{rd}$ barriers, respectively. For the 281 mg/L (4.5 mM) plume, nitrate concentrations decreased by ~1% (0.02 mM), ~6% (0.27 mM), and ~8% (0.37 mM) after $1^{st}$, $2^{nd}$, and $3^{rd}$ barriers, respectively. Nitrate plume was flowed through the flow-tank for 49 days by supplying $1.24\;m^3/d$ of nitrate solution. During nitrate treatment, flow velocity (0.44 m/d), pH (6.7 to 8.3), and DO (0.9~2.8 mg/L) showed little variations. Incomplete destruction of nitrate plume was attributed to the lack of retention time, rarely transverse dispersion, and inhibiting the activity of denitrification enzymes caused by relatively high DO concentrations. For field applications, it should be considered increments of retention time, modification of well placements, and intrinsic DO concentration.