• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.408-413
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• 2006

A strain, P821, with phospholipase D activity was isolated from soil and identified as a Streptomyces species. The phospholipase D enzyme was purified from a culture broth of the isolated strain using ammonium sulfate precipitation and DEAE-Sepharose, phenyl-Sepharose, and Superose 12 HR column chromatographies. The purified enzyme exhibited an optimum temperature and pH of $55^{\circ}C$ and 6.0, respectively, in the hydrolysis of phosphatidylcholine and remained stable up to $60^{\circ}C$ within a pH range of 3.5-8.0. The enzyme also catalyzed a transphosphatidylation reaction to produce phosphatidylserine with phosphatidylcholine and serine substrates. The optimum conditions for the transphosphatidylation were $30^{\circ}C$ and pH 5.0, indicating quite different optimum conditions for the hydrolysis and transphosphatidylation reactions. The gene encoding the enzyme was cloned by Southern hybridization and colony hybridization using a DNA probe designed from the conserved regions of other known phospholipase D enzymes. The resulting amino acid sequence was most similar to that of the PLD enzyme from Streptomyces halstedii (89.5%). Therefore, the enzyme was confirmed to be a phospholipase D with potential use in the production of phosphatidylserine.

• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.1018-1023
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• 2001

Oleamide (cis-9-octadecenamide) is endogenous primary amide of fatty acid that is produced in small amounts in animal brains. It is known to induce sleep and to lower temperature by destroying the lipid plasma membrane structure of cells, thereby disclosing gap junction channels. To develop a new biological production method for oleamide, a screening program was conducted to isolate a microorganism producing oleamide. Among 1,500 soil microorganisms tested, KK90378 exhibited a potent positive reaction with Dragendoff`s reagent, used to detect the primary amide of oleamide. KK90378 was identified as a Streptomyces species based on cultural and morpohological characteristics, the presence of diaminopimelic acid in the cell wall, and the sugar patterns for the whole-cell extrat. Streptomyces sp. KK90378 produced oleamide 3 days after culture at $28^{\circ}C$, pH 7.2 A series of purification steps, including hexane extraction, silica gel column, and preparative thin layer chromatographies, were performed for the purification of oleamide. A spectrophotometric analysis using $^1H$, $^13C$-NMR, and GC-MS confirmed that the chemical structure of the purified oleamide was identical to that of authentic oleamide.

• Applied Biological Chemistry
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• 제37권2호
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• pp.110-114
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• 1994

In searching for new insecticidal compounds from soil microorganisms, strains of streptomyces species showed insecticidal activities on Musca domestica and Bombyx mori were selected. Compounds I-IV, which were isolated from the metabolites of no. 46 strain, were identified as piericidin $C_1$, $C_2$, $C_3$ and $D_1$, respectively by UV and NMR data analyses.

• Journal of the Korean Society of Food Science and Nutrition
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• 제38권6호
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• pp.801-806
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• 2009

A species producing transglutaminase (EC 2.3.2.13) was isolated from forest soil and identified as Streptomyces platensis YK-2. The transglutaminase was purified from culture broth by 50% methanol precipitation, followed by successive chromatography on DEAE-Sephadex. The yield and purification-fold was 63.4% and 2.2-fold, respectively. The purified microbial transglutaminase (MTG) migrated as a single band of approximately 45 kDa upon sodium dodecyl sulfate polyacrylamide gel eletrophoresis. The isoelectric point determined by multichambered electrofocusing was pH $6.0{\sim}7.0$. The enzyme was strongly inhibited by $Hg^{++}$, but was activated by $Cd^{++}$, $Mg^{++}$, $Mn^{++}$, $Pb^{++}$ and reducing agents such as dithiothreitol and mercaptoethanol.

• Journal of Microbiology and Biotechnology
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• 제31권3호
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• pp.464-474
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• 2021

Bacterial cytochrome P450 (CYP) enzymes are responsible for the hydroxylation of diverse endogenous substances with a heme molecule used as a cofactor. This study characterized two CYP154C3 proteins from Streptomyces sp. W2061 (CYP154C3-1) and Streptomyces sp. KCCM40643 (CYP154C3-2). The enzymatic activity assays of both CYPs conducted using heterologous redox partners' putidaredoxin and putidaredoxin reductase showed substrate flexibility with different steroids and exhibited interesting product formation patterns. The enzymatic characterization revealed good activity over a pH range of 7.0 to 7.8 and the optimal temperature range for activity was 30 to 37℃. The major product was the C16-hydroxylated product and the kinetic profiles and patterns of the generated hydroxylated products differed between the two enzymes. Both enzymes showed a higher affinity toward progesterone, with CYP154C3-1 demonstrating slightly higher activity than CYP154C3-2 for most of the substrates. Oxidizing agents (diacetoxyiodo) benzene (PIDA) and hydrogen peroxide (H2O2) were also utilized to actively support the redox reactions, with optimum conversion achieved at concentrations of 3 mM and 65 mM, respectively. The oxidizing agents affected the product distribution, influencing the type and selectivity of the CYP-catalyzed reaction. Additionally, CYP154C3s also catalyzed the C-C bond cleavage of steroids. Therefore, CYP154C3s may be a good candidate for the production of modified steroids for various biological uses.

• Journal of Species Research
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• 제13권4호
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• pp.353-366
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• 2024

Here we describe indigenous prokaryotic species in Korea, a total of 41 bacterial strains were isolated from freshwater from the Republic of Korea. From the high 16S rRNA gene sequence similarity (>98.7%) and formation of a robust phylogenetic clade with the closest species, it was determined that each strain belonged to predefined bacterial species. There is no official report that these 41 species belonged to 5 phyla, 10 classes, 18 orders, 23 families, and 29 genera, which were assigned to Streptomyces, Mycolicibacterium, Smaragdicoccus, Nocardiopsis, and Nocardia of the phylum Actinobacteria; Runella, Flavobacterium, Algoriphagus, Sphingobacterium, and Aequorivita of the phylum Bacteroidota; Paenibacillus, Bacillus, Metabacillus, and Fredinandcohnia of the phylum Firmicutes; Sphingobium, Erythrobacter, Duganella, Methylopila, Novosphingobium, Azospirillum, Simplicispira, Corallococcus, Pseudomonas, Devosia, Pseudorhodoferax, Pseudomonas, Prolinoborus, Pectobacterium, and Aquabacterium of the phylum Proteobacteria; Proshecobacter of the phylum Verrucomicrobia. Gram reaction, colony and cell morphology, basic biochemical characteristics, isolation source, and strain IDs are also described in the species description section.

• Journal of Korean Society of Environmental Engineers
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• 제32권3호
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• pp.227-233
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• 2010

This study examined microbial community structures (MCSs) according to environmental factors through DGGE analysis and comparison in various soils collected from near army bases and gas stations. As a result, the similarities based on DGGE band profiles showed the closer relationship in regional properties than in pollution characteristics, probably due to the degree of weak contamination. The highly contaminated samples with oil revealed low MCS similarities with others in the same region and very low with all the other samples in the other regions. Thus the microbial community structure would more be affected by region-based natural factors than by contamination factors in case of minor pollution. All the dominant culturable bacterial species were involved in firmicutes or high GC Gram+ in a major portion of soil samples and the highly oil-contaminated samples contained Arthrobacter, Bacillus, Methylobacterium, Clavibacter, Streptomyces and Nocardia as reported genera, and Leifsonia as a unreported genus.

• Korean Journal of Microbiology
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• 제41권2호
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• pp.140-145
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• 2005

Streptomyces natalensis ATCC27448 produces natamycin, a commercially important macrolide antifungal antibiotic. For molecular genetic study of S. natalensis, we have developed a system for introducing DNA into S. natalensis via conjugal transfer from Escherichia coli. An effective transformation procedure for S. natalensis was established based on transconjugation from E, coli ET12567/pUZ8002 using a ${\Phi}C31$-derived integration vector, pSET152, containing oriT and attP fragments. The high frequency was obtained on MS medium containing 10 mM $MgCl_2$ using $6.25\times10^8$ of E.coli donor cells without heat treatment of spores. In addition, southern blot analysis of exconjugants and the sequence of plasmids containing DNA flanking the insertion sites from the chromosome revealed that S. natalensis contains a single ${\Phi}C31$ attB site and at least a secondary or pseudo attB site. Similar to the case of various Streptomyces species, a single ${\Phi}C31$ attB site of S. natalensis is present within an ORF encoding a pirin-homolog, but a pseudo-attB site is present within a distinct site (GenBank accession no. $YP\_117731$) and also its sequence deviates from the consensus sequences of attB sequence.

• Journal of Species Research
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• 제12권4호
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• pp.267-276
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• 2023

To obtain unrecorded bacterial species in Korea, various soils of coastal areas were collected from the Republic of Korea in 2022. After plating the samples on marine agar and incubating aerobically and anaerobically, approximately 1,700 bacterial strains were isolated and identified using 16S rRNA gene sequences. A total of 20 strains showed ≥98.7% 16S rRNA gene sequence similarity with validly published bacterial species but not reported in Korea, indicating they are unrecorded bacterial species in Korea. The unrecorded bacterial strains belonged to four phyla, six classes, 15 orders, 16 families, and 19 genera which were assigned to Blastomonas and Sphingomonas of the class Alphaproteobacteria; Pseudidiomarina, Kushneria, Salinicola, and Salinisphaera of the class Gammaproteobacteria; Evansella, Virgibacillus, and Paenibacillus of the class Bacilli; Cyclobacterium of the class Cytophagia; Pedobacter of the class Sphingobacteriia; and Demequina, Ornithinimicrobium, Blastococcus, Jatrophihabitans, Kineococcus, Glaciihabitans, Aeromicrobium and Streptomyces of the class Actinomycetes. The details of the 20 unreported species, including Gram reaction, morphology, biochemical characteristics, and phylogenetic position are also provided in the description of the strains.

• The Korean Journal of Mycology
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• 제31권1호
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• pp.44-49
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• 2003

Several microbial biocontrol products (Greenbiotech Co., Paju, Korea), Green-all T (Trichoderma harzianum), Green-all S(Bacillus sp.) and Green-all G (Streptomyces sp.) were supplemented with organic amendments such as sawdusts and rice hulls to study on efficacy of biological control of seeding damping-off of cucumber caused by Pythium ultimum. Sawdusts amended into potato dextrose agar alone could inhibit in vitro mycelial growth of P. ultimum. All there microbial products of Green-all T, Green-all G and Green-all S significantly reduced seeding damping-off (LSD, P=0.05). However, several amendments such as sawdusts and rice hulls into Green-all T and Green-all S products did not increase efficacy of biological control compared to non-amended treatment. In contrast, supplements of aminodoctor containing several amino acids (Greenbiotech Co., Korea) into Green-all G product significantly increased efficacy of biological control of seeding damping-off, resulting in from 42% to 2% disease incidence in relation to seedling emergence (LSD, P=0.05). Also, amendment of sawdusts into Tricoderma product significantly increased efficacy of biological control as disease index of 5.0 compared to non-amended control of 56.0 in Green-all T product alone. This indicates that organic amendments could increase efficacy of biological control of cucumber seedling damping-off.