• Mycobiology
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• 제39권3호
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• pp.174-181
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• 2011

The purpose of the present study was to investigate the influence of cultural and environmental parameters affecting the growth and bioactive metabolite production of the rare strain VUK-10 of actinomycete Pseudonocardia, which exhibits a broad spectrum of in vitro antimicrobial activity against bacteria and fungi. Production of bioactive metabolites by the strain was high the in modified yeast extract-malt extract-dextrose (ISP-2) broth, as compared to other tested media. Glucose (1%) and tryptone (0.25%) were found to be the most suitable carbon and nitrogen sources, respectively, for optimum production of growth and bioactive metabolites. Maximum production of bioactive metabolites was found in the culture medium with initial pH 7 incubated with the strain for four days at $30^{\circ}C$, under shaking conditions. This is the first report on the optimization of bioactive metabolites by Pseudonocardia sp. VUK-10.

• Journal of Microbiology and Biotechnology
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• 제25권5호
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• pp.629-636
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• 2015

Chemical investigation of the actinobacterial isolate Pseudonocardia endophytica VUK-10 has led to the segregation of two known bioactive compounds, namely 4-(2-acetamidoethyl) phenyl acetate and 4-((1, 4-dioxooctahydropyrrolo [1, 2-a] pyrazin-3-yl) methyl) phenyl acetate. The strain was isolated from a sediment sample of the Nizampatnam mangrove ecosystem, south coastal Andhra Pradesh, India. The chemical structure of the active compounds was established on the basis of spectroscopic analysis, including ¹H NMR and ¹³C NMR spectroscopies, FTIR, and EIMS. The antimicrobial and cytotoxic activities of the bioactive compounds produced by the strain were tested against opportunistic and pathogenic bacteria and fungi and on MDA-MB-231, OAW, HeLa, and MCF-7 cell lines. The compounds exhibited antimicrobial activities against gram-positive and gram-negative bacteria and fungi and also showed potent cytotoxic activity against MDA-MB-231, OAW, HeLa, and MCF-7 cell lines. This is the first example for this class of bioactive compounds isolated from Pseudonocardia of mangrove origin.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권5호
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• pp.408-413
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• 2006

We assessed the ability of a Pseudonocardia sp. from soil samples to bioconvert vitamin $D_3$. The optimal culture conditions for the bioconversion of vitamin $D_3$ to active $1{\alpha}$,25-dihydroxyvitamin $D_3$ were investigated by varying the carbon and nitrogen sources, the metal salt concentrations, the initial pH, and the temperature. Microbial transformations were carried out with the addition of vitamin $D_3$ dissolved in ethanol. They were sampled by extraction with methanol-dichloromethane and the samples were examined by HPLC. Optimum culture conditions were found to be 0.4% yeast extract, 1% glucose, 3% starch, 1% fish meal, 0.2% NaCl, 0.01% $K_2HPO_4$, 0.2% $CaCO_3$, 0.01% NaF, and pH 7.0 at $28^{\circ}C$. The optimal timing of the addition of vitamin $D_3$ for the production of calcitriol by Pseudonocardia autotrophica ID9302 was concurrent with the inoculation of seed culture broth. Maximum calcitriol productivity and the yield of bioconversion reached a value of 10.4mg/L and 10.4% respectively on the 7th day in a 75L fementer jar under the above conditions.

• 미생물학회지
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• 제40권3호
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• pp.221-225
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• 2004

본 논문에서는 희소 방선균, Pseudonocardia autotrophica(KCTC 9441) 유래 신규 Cytochrome P450 hydroxylase(CYP) 유전자들을 분리하여 염기서열 특성을 규명하였으며, 기존에 밝혀진 다른 방선균 유래 CYP 유전자들과의 연관성을 알아보았다. 이를 위하여 P. autotrophica의 cosmid DNA library를 제작하였고, 방선균에서 발견되는 CYP 유전자군의 보존된 서열로부터 제작된 degenerate primers를 이용한 PCR을 수행하여, P. autotrophica cosmid DNA library를 검색하였다. P. autotrophica cosmid DNA library검색 결과, P. autotrophica에는 염기서열이 서로 다른 4종의 신규 CYP유전자가 존재함이 확인되었으며 (CYP601-1, 601-2, 602, 605), 이들 신규 CYP유전자들은 방선균 유래 2차대사산물의 생합성에 관여하는 CYP유전자와 높은 유사성을 나타냈다. 특히, pESK601에서 확인된 CYP 유전자 및 주변 유전자의 염기서 열을 검색한 결과, polyene 계열의 항진균제, amphotericin과 nystatin의 생합성 유전자들과 매우 높은 유사성을 보임으로써, P. autotrophica에는 신규polyene계열의 항진균제 화합물의 생합성 유전자 군이 존재함도 규명되었다.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1226-1231
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• 2014

The rare actinomycete Pseudonocardia autotrophica was previously shown to produce a solubility-improved toxicity-reduced novel polyene compound named $\underline{N}ystatin$-like $\underline{P}seudonocardia$ $\underline{P}olyene$ (NPP). The low productivity of NPP in P. autotrophica implies that its biosynthetic pathway is tightly regulated. In this study, $wblA_{pau}$ was isolated and identified as a novel negative regulatory gene for NPP production in P. autotrophica, which showed approximately 49% amino acid identity with a global antibiotic down-regulatory gene, wblA, identified from various Streptomycetes species. Although no significant difference in NPP production was observed between P. autotrophica harboring empty vector and the S. coelicolor wblA under its native promoter, approximately 12% less NPP was produced in P. autotrophica expressing the wblA gene under the strong constitutive $ermE^*$ promoter. Furthermore, disruption of the $wblA_{pau}$ gene from P. autotrophica resulted in an approximately 80% increase in NPP productivity. These results strongly suggest that identification and inactivation of the global antibiotic down-regulatory gene wblA ortholog are a critical strategy for improving secondary metabolite overproduction in not only Streptomyces but also non-Streptomyces rare actinomycete species.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.396-397
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• 2005

The polyene antibiotics are a family of most promising antifungal polyketide compounds, typically produced by actinomycetes species. Using the polyene CYP-specific PCR screening with served actinomycetes genomic DNAs, Pseudonocardia autotrophica strain was identified to contain a unique polyene-specific CYP gene. The genomic DNA library screening using the polyene-specific CYP gene probe revealed the positive cosmid clone containing an approximately 34.5 kb DNA fragment revealed a total of seven complete and two incomplete open reading frame (ORFs), which are highly homologous but unique to previously-known polyene biosynthetic genes. These results suggest that the polyene-specific screening approach should be an efficient way of isolating potectially-valuable cryptic polyene biosynthetic gene cluster from various rare actinomycetes.

• 한국초전도ㆍ저온공학회논문지
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• 제25권3호
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• pp.7-12
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• 2023

Isolation of pollutant-degrading bacteria is important in bioaugmentation, one of the methods for biological degradation of environmental contaminants. We focused on the magnetic activated sludge (MAS) process as a culture method that efficiently concentrates degrading bacteria, and cultured activated sludge with 1,4-dioxane as a model pollutant. After 860 days of operation, MLVSS, which indicates the amount of sludge, increased from 390 mg/L to 10,000 mg/L, and the removal rate of organic matter including 1,4-dioxane, tetrahydrofuran, and glucose in the artificial wastewater reached up to 97%. Based on these results, the MAS process was successfully used to acclimate activated sludge with 1,4-dioxane. Bacterial flora analysis in the MAS showed that bacteria of the genus Pseudonocardia, already reported as 1,4-dioxane degrading bacteria, play an important role in the degradation of this pollutant. The MAS process is a suitable culture method for acclimation of environmental pollutants, and the findings indicate that it can be used as an enrichment unit for pollutant-degrading bacteria.

• Journal of Microbiology
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• 제40권3호
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• pp.211-218
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• 2002

A 4.8-kb DNA fragment encoding the P-450 type hydroxylase and ferredoxin genes was cloned from Pseudonocardia autotrophica IFO 12743 that can convert vitamin D$\_$3/ into its hydroxylated active forms. In order to isolate the P-450 gene cluster in this organism, we designed PCR primers on the basis of the regions of an oxygen binding site and a heme ligand pocket that are general characteristics of the P-450 hydroxylase. Sequencing analysis of the BamHI fragment revealed the presence of four complete and one incomplete ORFs, named PauA, PauB, PauC, and PauD, respectively. As a result of computer-based analyses, PauA and PauB have homology with enoyl-CoA hydratase from several organisms and the positive regulators belonging to the tetR family, respectively. PauC and PauD show similarity with SuaB/C proteins and ferredoxins, respectively, which are composed of P-450 monooxygenase systems for metabolizing two sulfonylurea herbicides in Streptomyces griseolus PauC shows the highest similarity with another CytP-450$\_$Sca2/ protein that is responsible for production of a specific HMG-CoA reductase inhibitor, pravastatin, in S. carbophilus. Cultures of Steptomyces lividans transformant, containing the P-450 gene cluster on the pWHM3 plasmid, was unable to convert vitamin D$\_$3/ to its hydroxylated forms.

• 농약과학회지
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• 제21권1호
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• pp.1-8
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• 2017

유기염소계 살충제 엔도설판은 2012년 국제적으로 사용이 금지되었으나 인도와 중국을 비롯한 국가들에서 여전히 사용되고 있으며 높은 잔류성과 이동성으로 인해 다양한 환경에서 검출되고 있다. 본 연구에서는 엔도설판이 토양세균의 군집구조에 미치는 영향을 16S rRNA 유전자 파이로시퀀싱 기법을 이용하여 분석하였다. 엔도설판을 100 mg/kg의 수준으로 밭토양에 처리했을 때 세균의 Operational taxonomic unit (OTU) 수와 다양성 지수가 감소했다가 서서히 증가하였으며 Proteobacteria와 Verrucomicrobia 문의 점유율은 증가하고 Chloroflexi와 Spirochaetes 문의 점유율은 감소한 것으로 나타나 엔도설판 처리가 토양세균 군집구조를 변화시키는 것을 확인하였다. 엔도설판 처리 시 Sphingomonas와 Burkholderia 속에 속하는 OTU의 점유율이 배양 초기에 높게 나타났으며 Pseudonocardia와 Opitutus 속에 속하는 OTU의 점유율은 배양 후기에 높게 나타났다. 이전 연구에서 Sphingomonas, Burkholderia, Pseudonocardia 속에 속하는 균주들이 엔도설판을 분해하였으므로 이 속에 속하는 OTU들이 토양에서 엔도설판의 분해에 단계적으로 관여했을 것으로 추정된다.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2005년도 2005 Annual Meeting & International Symposium
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• pp.331-331
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• 2005