• Molecules and Cells
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• v.37 no.10
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• pp.727-733
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• 2014

Spinosyns A and D are potent ingredient for insect control with exceptional safety to non-target organisms. It consists of a 21-carbon tetracyclic lactone with forosamine and tri-Omethylated rhamnose which are derived from S-adenosyl-methionine. Although previous studies have revealed the involvement of metK1 (S-adenosylmethionine synthetase), rmbA (glucose-1-phosphate thymidylyltransferase), and rmbB (TDP-D-glucose-4, 6-dehydratase) in the biosynthesis of spinosad, expression of these genes into rational screened Saccharopolyspora spinosa (S. spinosa MUV) has not been elucidated till date. In the present study, S. spinosa MUV was developed to utilize for metabolic engineering. The yield of spinosyns A and D in S. spinosa MUV was $244mgL^{-1}$ and $129mgL^{-1}$, which was 4.88-fold and 4.77-fold higher than that in the wild-type ($50mgL^{-1}$ and $27mgL^{-1}$), respectively. To achieve the better production; positive regulator metK1-sp, rmbA and rmbB genes from Streptomyces peucetius, were expressed and co-expressed in S. spinosa MUV under the control of strong $ermE^*$ promoter, using an integration vector pSET152 and expression vector pIBR25, respectively. Here-with, the genetically engineered strain of S. spinosa MUV, produce spinosyns A and D up to $372/217mgL^{-1}$ that is 7.44/8.03-fold greater than that of wild type. This result demonstrates the use of metabolic engineering on rationally developed high producing natural variants for the production.

• The Plant Pathology Journal
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• v.15 no.1
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• pp.14-20
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• 1999

Twenty isolates of Helminthosporium species were obtained from various grass plants and tested for controlling efficacy on the development of plant diseases. An isolate of Helminthosporium sp. TP-4 was chosen and six antibiotic substances were purified from cultures of the fungus by repeated silica gel column chromatography and preparative thin-layer chromatography. They were identified as ophiobolin a, 6-epiophiobolin A, 3-anhydroophiobolin A, 3-anhydro-6-epiophiobolin A, iphiobolin B, and iphiobolin I mainly by mass spectrometry and nuclear magnetic resonance spectrometry. Ophiobolins inhibited the growth of a grampositive bacterium Streptomyces griseus, but were not active against gram-negative bacteria. They also showed an antifungal activity. In in vivo tests, iphiobolin B exhibited potent controlling activities against rice blast, tomato late blight, and wheat leaf rust with control values more than 90% and 70% at concentration of $500\mu\textrm{m}$/ml and 100 ${\mu}{\textrm}{m}$/ml. Ophiobolin A and 6-epiophiobolin A controlled the development of wheat leaf rust more than 80% at concentrations of 100 /ml and $500\mu\textrm{m}$/ml respectively. 3-Anhydro-6-epiophiobolin A was not active against any plant disease. On the other hand, the A-series ophiobolins other than 3-anhydroophiobolin A showed stronger phytotoxic activity in a leaf-wounding assay using 8 plant species than those of 3-anhydroophiobolin A, ophiobolin B, and ophiobolin I. The results indicate that there is little correlation between antifungal activity and phytotoxicity of ophiobolins.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.1067-1070
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• 2004

In the course of screening for specific inhibitors against the mycelial form of Candida albicans from natural resources, we have isolated and identified A6792-1 from Streptomyces sp. A6792 by using several chromatographies. By spectral analyses, this compound was determined as 7-oxostaurosporine, having a structure of staurosporine aglycon noiety. 7-Oxostaurosporine exhibited a selective growth inhibitory activity against the mycelial form of Candida spp. up to $100\mu\textrm{g}/disc$ in bioassay. It also exhibited a specific antifungal activity against the mycelial form of Candida spp. including C. krusei, C. albicans, C. tropicalis, and C. lusitaniae with MICs ranging from 3.1 to $25\mu\textrm{g}/ml$ 7-Oxostaurosporine demonstrated no in vivo toxicity in SPF ICR mice. Therefore, this compound may have a considerable potential as an antifungal agent based on the preferential inhibition against growth of the mycelial form of Candida spp., dimorphic fungi.

• Microbiology and Biotechnology Letters
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• v.7 no.2
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• pp.97-102
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• 1979

An actinomycete, AS-568, which produced an inhibitory substance against succinate dehydrogenase of Ehrlich ascites carcinoma and Sarcoma-180 cells of mouse, was isolated. The inhibitory activity was determined by SDI (Succinate Dehydrogenase Inhibition) method. The active substance was specific against carcinoma cells compared to normal cells in mouse; liver, kidney and brain. The inhibitory ratio was about 50％ after one hr treatment at 37$^{\circ}C$ in vitro. Maximal productivity of active substance was recognized by 5 days culture in glucose-asparagine. The active component in cultural liquid was stable in neutral pH range and heat treatment reasonably, add it was recovered from precipitate by ammonium sulfate or non-dialyrable fraction in cellophane membrane as showing the behavior of high molecular substance.

• Journal of Microbiology and Biotechnology
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• v.17 no.11
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• pp.1856-1861
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• 2007

Physiological cell conditions such as glucose deprivation and hypoxia play roles in the development of drug resistance in solid tumors. These tumor-specific conditions cause decreased expression of DNA topoisomerase $II{\alpha}$, rendering cells resistant to topo II target drugs such as etoposide. Thus, targeting tumor-specific conditions such as a low glucose environment may be a novel strategy in the development of anticancer drugs. On this basis, we established a novel screening program for anticancer agents with preferential cytotoxic activity in cancer cells under glucose-deprived conditions. We recently isolated an active compound, AA-98, from Streptomyces sp. AA030098 that can prevent stress-induced etoposide resistance in vitro. Furthermore, LC-MS and various NMR spectroscopic methods identified AA-98 as mithramycin, which belongs to the aureolic acid group of antitumor compounds. We found that mithramycin prevents the etoposide resistance that is induced by glucose deprivation. The etoposide-chemosensitive action of mithramycin was just dependent on strict low glucose conditions, and resulted in the selective cell death of etoposide-resistant HT-29 human colon cancer cells.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.135-135
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• 1998

To find an antagonist of Pythium ultimum, the causal agent of damping-off, numerous actinomycete strains were screened for in vitro inhibiting mycelial growth of the target fungus and producing bioactive metabolites. A strain identified as Streptomyces sp. G60655 was isolated and used for further antagonistic efficacy. The degree of antagonism between the fungus and G60655 was affected by the medium used. Furthermore, the preinoculation of the antagonist was found to be necessary to exhibit the maximum efficacy of antagonsim against the fungus. From the culture broth, a bioactive metabolite was detected and purified by solvent extraction, silica gel chromatography and preparative HPLC. The FAB-MS spectrum of the active compound showed a molecular ion peak at m/z 1101 (M + H)$\^$+/, suggesting the molecular weight of 1100. The UV absorptions at 242 and 323 nm indicated the presence of aromatic functions. The structure of this compound was identified as echinomycin, a depsipeptide antibiotic by spectroscopic studies including various NMR measurements. Echinomycin was inactive against several soil born fungi, but inhibited the mycelial growth of P. ultimum and its related oomycetous fungi.

• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.885-892
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• 2011

The bacterial communities in the intestinal tracts of earthworm were investigated by culture-dependent and -independent approaches. In total, 72 and 55 pure cultures were isolated from the intestinal tracts of earthworms under aerobic and anaerobic conditions, respectively. Aerobic bacteria were classified as Aeromonas (40%), Bacillus (37%), Photobacterium (10%), Pseudomonas (7%), and Shewanella (6%). Anaerobic bacteria were classified as Aeromonas (52%), Bacillus (27%), Shewanella (12%), Paenibacillus (5%), Clostridium (2%), and Cellulosimicrobium (2%). The dominant microorganisms were Aeromonas and Bacillus species under both aerobic and anaerobic conditions. In all, 39 DNA fragments were identified by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis. Aeromonas sp. was the dominant microorganism in feeds, intestinal tracts, and casts of earthworms. The DGGE band intensity of Aeromonas from feeds, intestinal tracts, and casts of earthworms was 12.8%, 14.7%, and 15.1%, respectively. The other strains identified were Bacillus, Clostridium, Enterobacter, Photobacterium, Pseudomonas, Shewanella, Streptomyces, uncultured Chloroflexi bacterium, and uncultured bacterium. These results suggest that PCR-DGGE analysis was more efficient than the culturedependent approach for the investigation of bacterial diversity and the identification of unculturable microorganisms.

• Journal of Microbiology and Biotechnology
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• v.28 no.8
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• pp.1332-1338
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• 2018

In the course of studies to discover natural products with anti-angiogenic properties, two cyclic octapeptides, octaminomycins A (1) and B (2), were isolated from the cultures of Streptomyces sp. RK85-270. Octaminomycins suppressed the vascular endothelial growth factor (VEGF)-induced proliferation, adhesion, tube formation, migration, and invasion of HUVECs. Anti-angiogenic activity was futher confirmed in vivo by the chicken chorioallantoic membrane assay. We also identified that 1 and 2 inhibited the phosphorylation of VEGF receptor 2, AKT, and ERK1/2 and the expression and activities of MMP-2 and MMP-9. These results suggest that 1 and 2 may serve as potential scaffolds for the development of therapeutic agents to angiogenesis-dependent diseases.

• Applied Biological Chemistry
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• v.38 no.6
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• pp.516-521
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• 1995

In order to find a potent anti-methicillin resistant Staphylococcus aureus (MRSA) antibiotic, actinomycetes isolated from the samples collected in Korean marine silt were screened. From the culture broth of the isolated Streptamyces strain AM045, a substance showing excellent biological activity against MRSA was found, isolated and named AM3. The compound showed strong activities against MRSA, S. epidermidis, E. faecium and E. faecalis, which were better than those of vancomycin and teicoplanin. Unfortunately, AM3 was identified as Actinomycin V. However, this paper reports the three dimensional study of AM3 based on high resolution nmr and Computer Aided Molecular Modeling(CAMM), and the fact that the structure of the pentapeptide lactone ring with oxo-proline in chloroform solution does not have 'C conformation' any more.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.35-38
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• 1990

$\alpha$-D-Glucosidase inhibitor purified in a pure form was amorphous powder which gave a single spot at Rf value 0.12-0.71 with various developing solvent systems on silica gel thin layer chromatography, and melting point was 154.3-155.3$^{\circ}C$. It was disolved in water, formic acid and ethylene glycol monoethyl ether, and was very high hygroscopic substance. Biochemical reaction of the substance was positive to phenol sulfuric acid, ninhydrin, silver nitrate-sodium hydroxide, but negative to DNS reagent. Acid hydrolysis gave fructose and acid as sole sugar and amino acid constituents respectively. Moelcular weight of the inhibitor was estimated to be 1,050 by Shphadex G-25 column chromatography.