• Journal of Microbiology
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• v.45 no.3
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• pp.262-267
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• 2007

During a screening program, an actinomycete strain isolated from the Egyptian soil was investigated for its potential to show antimicrobial activity. The identification of this isolate was performed according to spore morphology and cell wall chemo-type, which suggested that this strain is a streptomycete. Further cultural, physiological characteristics and the analysis of the nucleotide sequence of the 16S rRNA gene (1480 bp) of this isolate indicated that this strain is identical to Streptomyces violaceusniger (accession number EF063682) and then designated S. violaceusniger strain HAL64. In its culture supernatant, this organism could produce one major compound strongly inhibits the growth of Gram-positive but the inhibition of Gram-negative indicator bacteria was lower. The antibiotic was separated by silica gel column chromatography and then purified on a sephadex LH-20 column and finally the purity was checked by HPLC. The chemical structure of the purified compound was determined using spectroscopic analyses (molecular formula of $C_{33}H_{32}N_{2}O_{10}$ and molecular weight of 617.21) and found to be identical to the kosinostatin, a quinocycline antibiotic which is known to be produced by Micromonspora sp. TP-A0468 (Igarashi et al., 2002) and to quinocycline B isolated from Streptomyces aureofaciens (Celmer et al., 1958). Although the antibiotic is known, the newly isolated strain was able to produce the antibiotic as a major product providing an important biotechnological downstream advantage.

• The Korean Journal of Mycology
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• v.26 no.1 s.84
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• pp.78-85
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• 1998

During the screening of biological control agents for insect pests in the greenhouse, 70 dead insect-related fungi were isolated and tested insecticidal effect of their culture filtrates was tested. From this studies, CNAB-63 isolate showed strong control effects against the cotton aphids and the two-spotted spider mite as 65.19% and 77.55%, respectively. The insecticidal active compound of CNAB-63 isolate was purified from culture filtrate by silica gel chromotography, thin layer chromatography and HPLC. Purified active compound (CNAB) showed control effects against the two-spotted spider mite with 63.45% at $100\;{\mu}g/ml$ concentration. However, it did not exhibit antimicrobial activity against fungi and bacteria.

• Korean Journal of Agricultural Science
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• v.48 no.2
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• pp.343-351
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• 2021

Screening was conducted using 200 kinds of plant extracts to explore herbicide-activated components of plant origin. We separated and purified active substances and elucidated chemical structures using Ligularia stenocephala M., which has strong activity and has not yet been studied. When the solvent fractions of the leaves of Ligularia stenocephala M. were tested for their herbicidal activity, ethyl acetate and chloroform layer showed an inhibition rate of 95.2% and 94.1%, respectively. In particular, the chloroform layer exerted more than 50% herbicidal activity at 10 ppm. From the chloroform layer with the highest herbicidal activity, we isolated three herbicidal active compounds using stepwise chromatography, specifically silica gel or octadecyl silica (ODS) column chromatography, Sep-pak cartridges, and high performance liquid chromatography (HPLC). Based on the analysis of the active compounds using electron ionization mass spectroscopy (EI-MS), ¹H-NMR, and ¹³C-NMR, we identified the active compounds as euparin, 5,6-dimethoxy-2-isopropenylbenzofuran, and liguhodgsonal. When the herbicidal activity of the identified compounds was tested, euparin showed selective herbicidal activity for lettuce at 10^-3 M, and both liguhogsonal and 5,6-dimethoxy-2-isoprophenylbenzofuran exerted selective activity for rice and Echinochloa crus-galli.

• Korean Journal of Pharmacognosy
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• v.53 no.3
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• pp.125-132
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• 2022

In this study, we investigated anti-oxidative and anti-inflammatory efficacy, and identified their constituents from Brassica napus L. (Korean name: Yuchae) whole plant. Upon the anti-oxidative activities screening, the ethanol extract exhibited potent DPPH and ABTS⁺ radical scavenging activities. On the anti-inflammation studies using LPS-induced RAW264.7 cells, the extract inhibited the production of NO and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) effectively. To identify major constituents of B. napus extract, further purification was performed and led to isolation of two compounds; isorhamnetin 3,7-O-diglucoside(1) and isorhamnetin 3-O-glucoside(2). Quantitative analysis by high pressure liquid chromatography (HPLC) determined the flavonoid 1 as the major constituent. Isolated compounds showed DPPH radical scavenging effects and decreased NO levels without causing cell toxicities. These results indicate that the extract of Yuchae, a rich plant resource in Jeju Island, could be potentially applicable as an anti-oxidative and/or anti-inflammatory ingredients.

• Korean Journal of Veterinary Service
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• v.36 no.4
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• pp.311-320
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• 2013

The purpose of this study was to evaluate detecting methods and the relationship between tissues and blood for enrofloxacin and metabolic ciprofloxacin residues in broiler chickens. Two groups of broiler chickens were administrated via the drinking water with $50{\mu}g/mL$ and $100{\mu}g/mL$ of enrofloxacin for 5 days, respectively. The concentration of enrofloxacin and metabolic ciprofloxacin in tissues (muscle and kidney) and blood were measured during administration period (for 5 days) and withdrawal period (for 12 days) by high performance liquid chromatography (HPLC) method. Also, all samples were conducted for screening of residues by microbial method using E. coli for quinolone detection and immuno-chromatography method using Smart kit. The relationship between tissues (muscle and kidney) and blood for enrofloxacin and metabolic ciprofloxacin residues in broiler chickens was followed : The levels of enrofloxacin and metabolic ciprofloxacin residues in muscle and kidney were higher 2.9~3.2 folds, 3.6~3.8 folds more than the residues levels in blood, respectively. These results support we can predict the residues in muscle and kidney from the residues in blood. In comparison of detecting methods for antibiotic residues, microbial method using E. coli for quinolone detection and immuno-chromatography method using Smart kit could detect positive reaction at similar or lower concentration than violative concentration of enrofloxacin and metabolic ciprofloxacin in chicken tissues. These results support what two screening methods are useful for screening of quinolone detection in chickens.

• KSBB Journal
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• v.24 no.2
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• pp.163-169
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• 2009

Lovastatin (also known as Mevinolin, Mevacor, and Monacolin K), an inhibitor of the HMG-CoA reductase produced by Aspergillus terreus and other fungi, is used to reduce serum cholesterol levels in human beings. It is derived biosynthetically from two polyketides. One of these is a nonaketide that undergoes cyclization at a hexahydronaphthalene ring system, and the other is a simple diketide, 2-methylbutyrate. Two primer pairs were designed based on the amino acid sequences of lovastatin polyketide synthase and lovastatin diketide synthase for the PCR screening of lovastatin-producing strains. Among the seven selected strains, SJ-2 evidenced the highest level of lovastatin production in both liquid and solid cultures. Soybeans with SJ-2 were treated via 1 hour of heat shock at $30^{\circ}C$ for the mass production of lovastatin. The heat-treated soybeans were inoculated on rice bran and the koji extract was obtained after 15 days of incubation. It yielded the highest level of lovastatin production among the strains, and also evidenced 75% inhibition activity against HMG-CoA reductase. We developed an efficient PCR screening method for lovastatin-producing strains, using lovastatin biosynthesis genes.

• Microbiology and Biotechnology Letters
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• v.33 no.2
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• pp.96-105
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• 2005

For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.26 no.1
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• pp.163-186
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• 2000

We have previously screened 150 medicinal plants on the inhibition of elastase and found a significant inhibitory effects of the extracts of Areca catechu L. on the aging and inflammation against the skin tissues. To isolate and identify the compounds having biological activity, we was further purified by each of the solvent fractions, silica gel column chromatography, preparative TLC and reversed-Phase HPLC. Peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as Phenolic substance using various colorimetric methods, UV, and IR. $IC_{50}$/ values of phenolic substance purified from Areca catechu were 26.9 $\mu\textrm{g}$/$m\ell$ for porcine pancreatic elastase (PPE) and 60.8 $\mu\textrm{g}$/$m\ell$ for human neutrophil elastase (HNE). This Phenolic substance showed more potent activity than those of reference compounds, oleanolic acid (76.5 $\mu\textrm{g}$/$m\ell$ for PPE, 219.2 $\mu\textrm{g}$/$m\ell$ for HNE) and ursolic acid (31.0 $\mu\textrm{g}$/$m\ell$ for PPE, 118.6 $\mu\textrm{g}$/$m\ell$ for HNE). According to the Lineweaver-Burk Plots, the inhibition against both PPE and HNE by this phenolic substance was competitive with substrate. Phenolic substance from Areca catechu exhibited high free radical scavenging effect ($SC_{50}$/ : 6 $\mu\textrm{g}$/$m\ell$) and inhibited effectively hyaluronidase activity ($IC_{50}$/: 210 $\mu\textrm{g}$/$m\ell$). These results suggest that the Phenolic substance Purified from Areca catechu showed anti-aging effect by protecting connective tissue proteins.

• Applied Biological Chemistry
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• v.48 no.2
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• pp.173-177
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• 2005

Physiological functionalities of water and ethanol extracts from Basil were determined. The concentration of total phenolic compounds of the water and ethanol extracts were $286.0\;{\mu}g/ml$, $250.0\;{\mu}g/ml$. Antioxidant activities of Basil extracts were determined using 2,2'-azinobis (3-ethyl benzothiazoline-6-sulfonic acid) radical cations (ABTS), 2,2-diphenyl-1-picryl hydrazyl radicals (DPPH), antioxidant protection factor and thiobarbituric acid reactive substances. The total antioxidant activities of Basil extracts using ABTS were 96.8% in the water extracts and 94.7% in the ethanol extract, DPPH were 87.0%, 93.9%, PF were 0.69, 1.16 and TBARS were $0.2{\times}10^{-3}\;{\mu}M,\;0.6{\times}10^{-3}\;{\mu}M$. Angiotensin converting enzyme inhibitory activity and xanthine oxidase inhibitory activity of Basil were higher in ethanol extracts (99.7%, 100.0%) than those of water extracts (39.9%, 54.7%). Phenolic profiles in Basil extracts were analyzed using HPLC. The result was that among the 6 phenolics, rosemarinic acid was the highest in ethanol extracts.

• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.137-142
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• 1998

Helicobacter pylori is a Gram-negative bacterium which causes chronic gastritis and is associated with gastric ulcer, duodenal ulcer and gastric carcinoma. In the process of screening of antibacterial activities against H. pylori from soil microorganisms, fungus No. 60686 was isolated. After fermentation of No.60686, the antibacterial compound was isolated, purified and followed by extraction of mycelium with organic solvents, acetone and ethyl acetate, through silica gel chromatography, LH-20 gel chromatography and HPLC. As a result of the structural analyses of the compound by IR, $^1$H- and $^{13}$C-NMR, FAB/Mass spectrophotometer, the compound having the antimicrobial activity was identified as chaetoglobosin A ($C_{32}H_{36}N_2O_5$), a cytochalasan derivative. The antimicrobial activity of chaetoglobosin A was tested against Gram-positive and negative bacteria by paper disk method. Among the test strains of 9 Gram-positive bacteria and 18 Gram-negative bacteria containing 4 H. pylori strains, the growth of 4 H. pylori strains and 3 S. aureus strains (SG 511, 285 and 503) was only inhibited by chaetoglobosin A. Also it was shown that its growth inhibition against H. pylori strains was stronger than that against S. aureus strains at the treatment of the same concentration. Therefore it was concluded that chaetoglobosin A has a specific growth inhibition against H. pylori of the tested bacteria.