• Mycobiology
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• v.39 no.3
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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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• v.25 no.5
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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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• v.11 no.5
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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.

• Korean Journal of Microbiology
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• v.40 no.3
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• pp.221-225
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• 2004

Novel cytochrome P450 hydroxylase (CYP) genes were isolated and characterized from P. autotrophica cosmid DNA library using an actinomycete CYP-specific PCR product as a screening probe. The cosmids containing four unique CYP genes (pESK601, 602, 603, 604, 605) were identified, and the four CYP genes were completely sequenced to be homologous to other known Actinomycetes CYP genes involved in various secondary metabolic pathways. Among all novel actinomycete CYP genes found in P. autotrophica, the CYP genes present in pESK601 were revealed to be highly homologous to the CYP genes involved in polyene-type amphotericin and nystatin antibiotic biosynthesis. The nucleotide sequences of the CYP flanking region in pESK601 also revealed the polyene-type biosynthetic genes, implying the presence of a cryptic polyene-type antifungal biosynthetic gene cluster in P. autotrophica.

• Journal of Microbiology and Biotechnology
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• v.24 no.9
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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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• 2005.04a
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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.

• Progress in Superconductivity and Cryogenics
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• v.25 no.3
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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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• v.40 no.3
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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.

• The Korean Journal of Pesticide Science
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• v.21 no.1
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• pp.1-8
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• 2017

Although a global ban on the use of endosulfan, an organochloline insecticide, has taken effect in mid-2012, it has been still used in several countries, including India and China, and detected in diverse environments in the world due to its relative persistence and semi-volatility. In this study, the effect of endosulfan on soil bacterial community was investigated using 16S rRNA gene pyrosequencing method. When endosulfan was applied to an upland soil at a rate of 100 mg/kg soil (ES soil), the number of operational taxonomic units (OTU) and diversity indices for bacteria initially decreased and gradually recovered to the level of the non-treated soil (NT soil) during an eight-week incubation period. At bacterial phylum level, relative abundances of Proteobacteria and Verrucomicrobia were higher while those of Chloroflexi and Spirochaetes were lower in the ES soil than in the NT soil, suggesting that an endosulfan application affects the bacterial community structure in soil. In the ES soil, the relative abundances of the OTUs affiliated to the genera Sphingomonas and Burkholderia increased in the initial period of incubation while those affiliated to the genera Pseudonocardia and Opitutus increased in the late period of incubation. Because the first three genera contain bacterial strains reported to degrade endosulfan, they are expected to be involved in the degradation of endosulfan, probably one after another.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2005.06a
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• pp.331-331
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• 2005