• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1127-1132
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• 2012

Nargenicin $A_1$ is a 28-membered polyketide macrolide, with antibacterial activity against methicillin-resistant Staphylococcus aureus, produced by Nocardia sp. CS682. In this study, the production of nargenicin $A_1$ was improved by enhancing the supply of different biosynthetic precursors. In Nocardia sp. CS682 (KCTC11297BP), this improvement was ~4.62-fold with the supplementation of 30 mM methyl oleate, 4.25-fold with supplementation of 15mM sodium propionate, and 2.81-fold with supplementation of 15 mM sodium acetate. In Nocardia sp. metK18 and Nocardia sp. CS682 expressing S-adenosylmethionine synthetase (MetK), the production of nargenicin $A_1$ was improved by ~5.57-fold by supplementation with 30 mM methyl oleate, 5.01-fold by supplementation with 15 mM sodium propionate, and 3.64-fold by supplementation with 15 mM sodium acetate. Furthermore, supplementing the culture broth of Nocardia sp. ACC18 and Nocardia sp. CS682 expressing the acetyl-CoA carboxylase complex (AccA2 and AccBE) with 30 mM methyl oleate, 15 mM sodium propionate, or 15 mM sodium acetate resulted in ~6.99-, 6.46-, and 5.58-fold increases, respectively, in nargenicin $A_1$ production. Our overall results showed that among the supplements, methyl oleate was the most effective precursor supporting the highest titers of nargenicin $A_1$ in Nocardia sp. CS682, Nocardia sp. metK18, and Nocardia sp. ACC18.

• 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.

• 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.

• 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 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.