• 한국미생물·생명공학회지
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• 제32권3호
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• pp.282-285
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• 2004

The polyene antibiotics including nystatin, pimaricin, amphotericin and candicidin are a family of most promising antifungal polyketide compounds, typically produced by rare actinomycetes species. The biosynthetic gene clusters for these polyenes have been previously investigated, revealing the presence of highly homologous biosynthetic genes among polyene-producers such as polyketide synthase (PKS) and cytochrome P450 hydroxylase (CYP) genes. Based on amino acid sequence alignment among actinomycetes CYP genes, the highly-conserved regions specific for only polyene CYP genes were identified and chosen for degenerate PCR primers, followed by the PCR-screening with various actinomycetes genomic DNAs. Among tested several polyene non-producing actinomycetes strains, Pseudonorcardia autotrophica strain was selected based on the presence of PCR product with polyene-specific CYP gene primers, and then confirmed to contain a cryptic novel polyene hydroxylase gene in the chromosome. These results suggest that the polyene-specific hydroxylase gene PCR should be an efficient way of screening and isolating potentially-valuable cryptic polyene antibiotic biosynthetic genes from various microorganisms including rare actinomycetes.

• Journal of Applied Biological Chemistry
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• 제42권3호
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• pp.119-124
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• 1999

About two thirds of the naturally occurring antibiotics have been discovered from actinomycetes. Therefore, the probability of discovering further new antibiotics from actinomycetes is declining as many known metabolites are isolated repeatedly. However, various efforts leave been made in order to enhance the probability of discovering novel compounds. In the present study, we have developed new screening strategies based on the antibiotic biosynthetic pathway, and the genetic information, utilizing polymerase chain reaction. We have selected macrolide type polyketides. In order to divide the ansamycin group antibotic of macrolide type polyketides, we have selected 3-amino-5-hydroxybenzoic acid (AHBA) moiety which contains a biosynthetically unique structural element in the group as a target molecules. Oligonucleotide primers were designed to amplify DNA fragments of macrolide type polyketide synthase and AHBA synthase genes from fourteen actinomycetes species. This method was successfully applied to all three of the known macrolide type polyketide produccing actinomycetes tested. In addition, it also identified the presence of potential macrolide type polyketide producing genes from seven actinomycetes that were known to produce none of macrolide type polyketides, and AHBA biosynthetic genes in one actinomycetes. This technique is potentially useful for the screening of new antibiotices and cloning of their biosynthetic genes.

• 한국미생물·생명공학회지
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• 제28권3호
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• pp.129-133
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• 2000

Different actinomycete strains were isolated from natural lime caves of Ondal Chemongok Hwanseon and Yongyeon which are located at Kangwon or chungcheongbook province in Korea and were identified to the genus level. Soil sam-ples were collected at 6 sites inside and 2 sites outside of each natural lime cave, As the result the strains belonging to genus Streptomyces and rare actinomycetes were isolated at the average of 2.1 and 3.4 strains per g soil on inside cave whereas which were isolated at the 6.0 and 1.8 strains per g soil on outside cave. How-ever the generic distribution of Streptomyces and rare actinomycetes isolated from outside cave was quite dif-ferent from that of inside cave. It was shown that rare actinomycetes at natural lime caves is generally highly abundant than Streptomyces.

• 한국미생물·생명공학회지
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• 제27권6호
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• pp.515-517
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• 1999

From the soils collected seasonally at the 0.5~2cm and 50$\pm$1cm depths of riverside, different strains of actinomycetes were isolated and identified to the genus. At the 0.5~2cm depth, Streptomyces and rare actinomycetes were in total 73 and 62 strains, respectively. Streptomyces was approximately 2-fold more in spring and autumn than summer, and rare actinomycetes was at least 4-fold more in autumn and winter than spring. At the 50$\pm$1cm depth, Streptomyces and rare actinomycetes were in total isolated 53 and 41 strains, and these were at least 2-fold more diverse in autumn than spring, summer, and winter.

• 한국미생물·생명공학회지
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• 제22권5호
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• pp.561-563
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• 1994

For the more effective isolation of soil actinomycetes, we have developed HHV (Hair hydrolysate-vitamin) agar medium, containing hair as the sole source of carbon and nitrogen. The HHV agar medium was superior to other media such as colloidal chintin agar, glycerol-arginine agar and starch-casein-nitrate agar, and HV (humic acid-vitamin) agar. The maximum effect of this medium has been shown in hair dry weight 0.4 g/l medium. Of each soil sample, the greatestest number of actinomycetes was isolated from the potato annual planted soil among the tested samp- les. The genus of actinomycetes isolated from the potato annual planted soil sample was identified such 5 group as Stretomyces, Micromonospora, Microbispora, Nocardia and Saccharopolyspora.

• 대한수의학회지
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• 제22권1호
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• pp.27-29
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• 1982

A bacteriological survey was made for the aerabic Actinonmycetes from the vaginal exudates of apparently healthy dairy cows in Gyeongbug area. One hundred twenty isolated were made from 1,328 cows. Among them 30 isolates (25%) were identified as Nocardia asteriodes, 4 (3.3%) as Nocaradia brasiliensis, and 2 (1.7%) as Nocardia madurae. The remaining 84 isolates (70%) were identified as Streptomyces species.

• 약학회지
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• 제32권5호
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• pp.304-307
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• 1988

394 strains of soil microorganism were isolated from the Korean soil samples. The isolated strains were shake-cultured in oat-meal medium. The filtrates of the cultures were screened for the production of ${\alpha}-amylase$ inhibitors. Five strains were identified to produce ${\alpha}-amylase$ inhibitors. And these strains were identified as Actinomycetes.

• Mycobiology
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• 제48권1호
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• pp.37-43
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• 2020

A serious leaf disease caused by Colletotrichum dematium was found during the cultivation of Sarcandra glabra in Jingxi, Rong'an, and Donglan Counties in Guangxi Province, which inflicted huge losses to plant productivity. Biological control gradually became an effective control method for plant pathogens. Many studies showed that the application of actinomycetes in biological control has been effective. Therefore, it may be of great significance to study the application of actinomycetes on controlling the diseases caused by S. glabra. Strains of antifungal actinomycetes capable of inhibiting C. dematium were identified, isolated and screened from healthy plants tissues and the rhizospheres in soils containing S. glabra. In this study, 15 actinomycetes strains were isolated and among these, strains JT-2F, DT-3F, and JJ-3F, appeared to show antagonistic effects against anthracnose of S. glabra. The strains JT-2F and DT-3F were isolated from soil, while JJ-3F was isolated from plant stems. The antagonism rate of strain JT-2F was 86.75%, which was the highest value among the three strains. Additionally, the JT-2F strain also had the strongest antagonistic activity when the antagonistic activities were tested against seven plant pathogens. Strain JT-2F is able to produce proteases and cellulase to degrade the protein and cellulose components of cell walls of C. dematium, respectively. This results in mycelia damage which leads to inhibition of the growth of C. dematium. Strain JT-2F was identified as Streptomyces tsukiyonensis based on morphological traits and 16S rDNA sequence analysis.

• Journal of Microbiology and Biotechnology
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• 제8권3호
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• pp.280-283
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• 1998

We have cloned the RNA polymerase sigma-like factors from a wide range of actinomycetes by using specific primers with the polymerase chain reaction (PCR). The specific oligonucleotide primers were designed on the basis of amino acid sequences of conserved regions from HrdA, B, D of Streptomyces griseus as well as from the rpoD box of many eubacteria. The consensus sequences were from the rpoD box and helix-turn-helix motif involved in -35 recognition. The designed primers were successfully applied to amplify the DNA fragments of the hrd homolog genes from 8 different strains of actinomycetes which produce a wide variety of important antibiotics. The 480 bp of the DNA fragment was amplified from all 8 strains, and it was identified as a part of hrdA and hrdB as we designed. The deduced amino acid sequence of PCR-amplified DNA fragments were highly homologous to those of other known RNA polymerase sigma factors of S. griseus and Streptomyces aureofaciens. Therefore, this study with specifically designed primers will support rapid cloning of the RNA polymerase sigma factors which recognize different classes of promoters from actinomycetes, and it will also be helpful in understanding the relationship of promoters and sigma factors leading to heterogeneity of RNA polymerases in actinomycetes.

• 한국미생물·생명공학회지
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• 제22권2호
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• pp.222-225
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• 1994

Three kinds of pretreatment methods were used for selectivel isolation of soil actinomy cetes. One hundred and six strains were isolated on Bennet's agar and 114 strains were on humic acid-vitamin agar from 5 domestic soil samples. All of these isolates were identified to the genus level based on morphological and physiological characheristics. Among three methods, dry heat was most effective to isolate many different strains including rare actinomycetes. On genus, Bennet's agar was effective for selective isolation of Nocardia but bumic acid-vitamin agar for another rare actinomycetes.