• Journal of Microbiology and Biotechnology
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• v.33 no.7
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• pp.949-954
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• 2023

Type III polyketide synthase (PKS) found in bacteria is known as 1,3,6,8-tetrahydroxynaphthalene synthase (THNS). Microbial type III PKSs synthesize various compounds that possess crucial biological functions and significant pharmaceutical activities. Based on our sequence analysis, we have identified a putative type III polyketide synthase from Nocardia sp. CS682 was named as ThnA. The role of ThnA, in Nocardia sp. CS682 during the biosynthesis of 1,3,6,8 tetrahydroxynaphthalene(THN), which is the key intermediate of 1-(α-L-(2-O-methyl)-6-deoxymannopyranosyloxy)-3,6,8-trimethoxynaphthalene (IBR-3) was characterized. ThnA utilized five molecules of malonyl-CoA as a starter substrate to generate the polyketide 1,3,6,8-tetrahydroxynaphthalene, which could spontaneously be oxidized to the red flaviolin compound 2,5,7-trihydroxy-1,4-naphthoquinone. The amino acid sequence alignment of ThnA revealed similarities with a previously identified type III PKS and identified Cys¹³⁸, Phe¹⁸⁸, His²⁷⁰, and Asn³⁰³ as four highly conserved active site amino acid residues, as found in other known polyketide synthases. In this study, we report the heterologous expression of the type III polyketide synthase thnA in S. lividans TK24 and the identification of THN production in a mutant strain. We also compared the transcription level of thnA in S. lividans TK24 and S. lividans pIBR25-thnA and found that thnA was only transcribed in the mutant.

• Korean Journal of Microbiology
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• v.25 no.1
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• pp.28-33
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• 1987

Celluloytic bacteria, -Gram positive, Gram negative and actionmycetes-were used to study their catabolic pathways of carbohydrate. It was found that Embden-Meyerhof-Parnas(EMP) pathway and hexose monophosphate(MHP) shunt were operated in Cellulomonase sp. CS1-1, C. flavigena, and Pseudomonas fluorescens subsp. cellulosa when they were cultured in a glucose containing medium, whilst gluconate was catabolised mainly via Entner-Doudoroff(ED) pathway, and to some extend through HMP shunt. Enzymes of ED pathway in the orgamisms were induced by gluconate. On the other hand Nocardia cellulans catabolised glucose and gluconate via EMP pathway and HMP shunt. The growth rate of N. Cellulans on gluconate were much slower than that on glucose.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 1996.11a
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• pp.114-117
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• 1996

Biodegradation potential of crude oil has been studied in liquid and soil slurry culture. Studies were performed to optimize the factors affecting metabolic activity. Arabian Light(sulfur content 1%) was used as a representative crude oil and Nocardia sp. was selected as an oil degrading microorganism based on its ability to degrade and emulsify Effects of various nutritional and environmental conditions as well as emulsification and surface tension were observed. Tentative optimization of environmental and nutritional condition were as follow; pH 8, sodium nitrate as inorganic nitrogen source, yeast extract 0.05%, phosphate concentration 0.25% and glucose addition of 1.0% (w/v basis), extent of degradation to 78 %.

• Journal of Microbiology and Biotechnology
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• v.19 no.4
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• pp.339-345
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• 2009

We evaluated the activity and abundance of the crude-oil-degrading bacterium Nocardia sp. H17-1 during bioremediation of oil-contaminated soil, using real-time PCR. The total petroleum hydrocarbon(TPH) degradation rate constants(k) of the soils treated with and without H17-1 were $0.103\;d^{-1}$ and $0.028\;d^{-1}$ respectively. The degradation rate constant was 3.6 times higher in the soil with H17-1 than in the soil without H17-1. In order to detect and quantify the Nocardia sp. H17-1 in soil samples, we quantified the genes encoding 16S ribosomal RNA(16S rRNA), alkane monooxygenase(alkB4), and catechol 2,3-dioxygenase(23CAT) with real-time PCR using SYBR green. The amounts of H17-1 16S rRNA and alkB4 detected increased rapidly up to 1,000-folds for the first 10 days, and then continued to increase only slightly or leveled off. However, the abundance of the 23CAT gene detected in H17-1-treated soil, where H17-1 had neither the 23CAT gene for the degradation of aromatic hydrocarbons nor the catechol 2,3-dioxygenase activity, did not differ significantly from that of the untreated soil($\alpha$=0.05,p>0.22). These results indicated that H17-1 is a potential candidate for the bioaugmentation of alkane-contaminated soil. Overall, we evaluated the abundance and metabolic activity of the bioremediation strain H17-1 using real-time PCR, independent of cultivation.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.901-905
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• 2004

The degradation and mineralization of crude oil were investigated over 50-days in three soils, loamy sand, sand, and combusted loamy, which were artificially contaminated with crude oil (50 g $kg^{-1}$) and inoculated with Nocardia sp. H17-1. The degradation efficiency of total petroleum hydrocarbon (TPH) in sand was the highest at 76% among the three soils. The TPH degradation rate constants $(k_{TPH})$ in loamy sand, sand, and combusted loamy sand were 0.027 $d^{-1}$, 0.063 $d^{-1}$, and 0.016 $d^{-1}$, respectively. In contrast, the total amount of $CO_2$ evolved was the highest at 146.1 mmol in loamy sand. The $CO_2$ evolution rate constants (k_{CO2})$ in loamy sand, sand, and combusted loamy sand were 0.057 $d^{-1}$, 0.066 $d^{-1}$, and 0.037 $d^{-1}$, respectively. Therefore, it seems that the degradation of crude oil in soils can be proportional to the soil pore space and that mineralization can be accelerated with the increase of organic substance.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.569-575
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• 2002

Two bacterial strains capable of degrading trichloroethylene (TCE), isolated form soils contaminated with various chlorinated alkenes, were identified as Alcaligenes odorous N6 and Nocardia sp. Hl7. In addition, four KCTC strains, including three strains of Pseudomonas putida and one strain of Sphingomonas chlorophenolica, exhibited an ability to degrade toluene. A. odorans N6 and Nocardia sp. H17 degraded 84% of the initial amount of TCE in a basal salts medium (BSM), containing 0.2 mM TCE as the sole source of carbon and energy, in a day. The optimal pH for growth was within a range of 7.0-8.0. A mixed culture of the four toluene-degrading isolates degraded 95% of 0.2 mM TCE with 1.5 mM toluene as an inducer, whereas no TCE was degraded by the same mixture without an inducer. When a mixed culture of all 6 isolates was used, the degradation efficiency of 0.2 mM TCE was 72% without an inducer, in a day, and 82% with toluene as an inducer. In a continuous treatment, 1,000 mg/1 of TCE in an artificial wastewater was completely removed within 18 h when an activated sludge was used along with the microbial mixture, which was 27 h laster than when only an activated sludge was used. Accordingly, it would appear that such a microbial mixture could be effectively applied to the biological treatment of wastewater containing TCE with or without an inducer.

• Korean Journal of Poultry Science
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• v.38 no.1
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• pp.71-80
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• 2011

This study was conducted to investigate the effects of dietary supplementation of CS682, a fermentation product of Actinomycetae Nocardia sp. CS682, and DCS682$^{(R)}$, a commercial product, on the performance, blood parameters, small intestinal microflora, and immunoglobulin contents in broilers. In Exp. 1, a total of 240 ROSS$^{(R)}$ broiler chickens of 1d old were assigned to six dietary treatments: Control, Antibiotics (6 ppm avilamycin), CS682-0.25 (CS682 0.25%), CS682-0.50, CS682-0.75 and CS682-1.00. There were significant (p<0.05) differences among treatments in feed conversion. The CS682-0.25 treatment was significantly (p<0.05) lower than Antibiotics and other CS682 treatments in 0~2 wk feed conversion. The CS682 treatments influenced MCV (mean corpuscular volume) in blood. The cfu of Escherichia coli in small intestinal content was lowest in Antibiotics treatment followed by CS682 treatments and Control. In Exp. 2, a total of 1,000 ROSS$^{(R)}$ broiler chickens of 1 d old were assigned to five dietary treatments: Control, Antibiotics (6 ppm avilamycin), DCS682-0.05 (DCS682$^{(R)}$ 0.05%), DCS682-0.10 and DCS682-0.20. There were significant differences (p<0.05) among treatments in mortality. The DCS682-0.20 treatment was lower than DCS682-0.10 in 0~3 wk and lower than Control in 0~5 wk mortality. Antibiotics treatment was lowest in all microbial population in small intestinal content. The cfu of E. coli and Salmonella typhimurium of DCS682 treatments were higher than Antibiotics treatment but lower than the Control. The results of present broiler experiments indicated that supplementation of 0.20~0.25% CS682 and DCS682, improve feed conversion, mortality and control harmful intestinal microbes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.11
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• pp.1507-1514
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• 2008

Cholesterol oxidase catalyses the conversion of cholesterol to 4-cholesten-3-one. This enzyme has been used for clinical assay of human serum cholesterol and for reduction of cholesterol level in foods and feeds. In order to search the microorganism which has a high extracellular and stable activity of cholesterol oxidase, soil microorganisms were screened. As a result, the one with the highest extracellular cholesterol oxidase activity was obtained and named as the BEN 115. The BEN 115 strain was identified as one of the Nocardia species based on our taxonomic studies. The cholesterol oxidase from this strain was shown to have two bands of extracellular proteins on SDS-PAGE and Western blot. Their molecular masses were estimated to be about 55 and 57 kDa, respectively. In addition, this cholesterol oxidase was considerably stable at the broad range of pH $3.5{\sim}9.5$ and at the temperature of $25{\sim}55^{\circ}C$. The optimum pH and temperature of this cholesterol oxidase were pH 5.5 and $35^{\circ}C$, respectively. The activity of extracellular cholesterol oxidase could be enhanced 1.6 to 2.0 folds by the addition of nonionic detergent such as Triton X-114, Triton X-100, or Tween-80 into the culturing broth. The substrate specificities against campesterol, sitosterol and stigmasterol were measured to be 50%, 50%, and 27%, respectively, compared to the cholesterol. These results suggest that Nocardia sp. BEN 115 may be useful as a microbial source of cholesterol oxidase production.

• Korean Journal of Poultry Science
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• v.38 no.1
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• pp.59-69
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• 2011

This study was conducted to investigate the effects of dietary supplementation of CS682, a fermentation product of Actinomycetae(Nocardia sp. CS682), and its commercial product DSC682$^{(R)}$ on the performance, blood parameters, intestinal microflora, and immune response in laying hens. Hy-Line Brown$^{(R)}$ laying hens were housed in two bird cages. Feeding trial lasted 5 wk under 16.5 h:7.5 h(L:D) lighting regimen. In Exp.1, a total of 480 birds of 86 wk old were assigned to four dietary treatments: Control, Antibiotics (6 ppm avilamycin), CS682-0.1 (CS682 0.1%) and CS682-1.0 (CS682 1.0% supplementation). Each treatment was replicated five times with 24 birds (or 12 cages) per replication. In Exp. 2, a total of 1,000 birds of 26 wk old were assigned to five dietary treatments: Control, Antibiotics (6 ppm avilamycin), DCS682-0.05 (DCS682 0.05%), DCS682-0.1 (DCS682 0.1%), DCS682-0.2 (DCS682 0.2% supplementation). Each treatment was replicated five times with 40 birds (or 20 cages) per replication. In Exp. 1, there were no significant differences among treatments in egg production, egg weight, broken & soft egg production, feed intake, and feed conversion ratio. Also, there were no significant differences among treatments in eggshell thickness, eggshell color and Haugh unit. However, eggshell strength was significantly (p<0.05) greater in CS682 and Antibiotics treatments than Control, and egg yolk color was significantly (p<0.05) higher in CS682-1.0 than Control. In Exp. 2, feed intake was significantly (p<0.05) lower in DSC682-0.05 than Control. Lightness(L) of Hunter Lab color of eggshell of DCS and Antibiotics treatments was significantly (p<0.05) lower than Control. Egg yolk color of DCS 0.1 and 0.2 treatments was significantly (p<0.05) higher than Control. Haugh unit increased significantly (p<0.05) in Antibiotics and DCS682-0.1 treatments. The immunoglobulin levels of plasma (IgG and IgA) and eggyolk (IgY) were not significantly affected by treatments. Antibiotics and CS682 or DCS682 treatments significantly (p<0.05 or 0.01) influenced some of the erythrocytes and leukocytes parameters in blood. In Exp.1, mean corpuscular volume (MCV) decreased by CS682 treatments and mean corpuscular hemoglobin (MCH) was highest in Antibiotics treatments. In Exp.2, the level of monocyte (MO) decreased in DCS682-0.10 and 0.20 treatments. The cfu of C. perfringens and S. typhimurium in small intestinal content were highest in Control and lowest in Antibiotics in both experiments. In Exp. 2, DSC682-0.05 and -0.1 treatments were highest and Antibiotic treatment was lowest in Lactobacilli spp. The results of the present layer experiments indicated that supplementation of 0.1~0.2% CS682 or DCS682 may increase eggshell strength, color of eggshell and eggyolk, Haugh unit, and control harmful intestinal microbes.

• Korean Journal of Organic Agriculture
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• v.22 no.3
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• pp.469-481
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• 2014

IIn vitro antimicrobial and anti-inflammatory activities of nargenicin and its derivatives were investigated. Nargenicin, an unusual macrolide antibiotic with potent anti-MRSA (methicilin-resistant Staphylococcus aureus) activity, was purified from the culture broth of Nocardia sp. CS682. And variety of novel nargenicin derivatives was synthesized from nargenicin. Two compounds (4 and 5) exhibit a broad spectrum of antimicrobial activities against infectious bacteria. The antimicrobial activity of derivatives against fifteen organisms was assessed using the minimum inhibitory concentration (MIC). The MIC values were in the ranges of $0.15{\sim}80{\mu}g/mL$ (w/v) for compound 1 and 2, $5{\sim}80{\mu}g/mL$ (w/v) for compound 3, $1.25{\sim}40{\mu}g/mL$ (w/v) for compound 4, and $1.25{\sim}80{\mu}g/mL$ (w/v) for compound 5, depending on the pathogens studied. In vitro, we investigated cytotoxicity and inhibition of nitric oxide (NO) production of synthesized compounds 1-5 in Raw 264.7 cells. LPS-induced nitric oxide releases were significantly blocked by compound 3, 4 and 5 in a dose-dependent manner. At high concentrations ($5{\mu}g/mL$) compound 5 inhibited the NO production by 95%. Compound 4 inhibited the release of NO in LPS-activated Raw 264.7 cells by 75% at the concentration of $10{\mu}g/mL$. Compound 3 inhibited the release of NO in LPS-activated Raw 264.7 cells by 65% at the concentration of $100{\mu}g/mL$. On the other hand, nargenicin, compound 1 and 2 did not inhibit NO production. These results demonstrated that compound 4 and 5 displayed antimicrobial activity and blocked LPS-induced pro-inflammatory mediators such as NO in macrophages, which might be responsible for its therapeutic application.