• Journal of Environmental Science International
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• v.30 no.5
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• pp.369-378
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• 2021

In this study, we investigated the effects of indole on biofilm formation inhibition in Pantoea agglomerans (P. agglomerans). In the biofilm growth assay, indole inhibited biofilm formation across all the growth time. Depending on biofilm growth stage, indole exhibited biofilm inhibition and anti-bacterial effects on planktonic cells. Through the analysis of the proportion rate between biofilm and Colony Forming Units (CFU) and inhibition rate of indole, we confirmed that depending on the biofilm stage of P. agglomerans, indole treatment timing was more important than the treatment duration. By comparing gene expression rates through rt-qPCR P.agglomerans affected by indole was found to significantly change quorum sensing (pagI/R) and indole transportation (bssS) gene expressions. Throughout all, indole exhibited both antimicrobial and anti-biofilm effects on P. agglomerans. In addition, we confirmed the anti-biofilm effects of indole on mature biofilm. In conclusion, indole as a signal molecule, can exhibit anti-biofilm effects through bacterial quorum sensing inhibition and indole affects. Therefore, indole can regulate biofilm bacteria especially gram-negative opportunistic pathogens.

• Microbiology and Biotechnology Letters
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• v.20 no.1
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• pp.73-78
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• 1992

- Enzymatic synthesis of L-tryptophan(Trp) using E. coli tryptophanase has been investigated. In order to reduce the substrate inhibition by indole and to increase the product yield of L-tryptophan three different approaches have been made in this work. First, indole was intermittently fed to the reaction mixture in order to control the indole concentration at lower level. When 15 mM of indole was used as a total amount of substrate, conversion yield of 80% has been obtained with intermittent feeding while only 20% of indole was converted into L-tryptophan by conventional batch operation, The second method employed in this work was the use of cyclohexane-phosphate buffer organic two-phase system. In this system, indole was mainly partitioned into the organic-solvent phase and therefore substrate inhibition was expected to be reduced. L-Tryptophan production in organic two-phase system was, however, unexpectedly lower than that obtained in aqueous buffer solution. As a third method cyclodextrins have been added to the aqueous reaction mixture. It was found that the addition of $\beta$-cyclodextrin enhanced the tryptophan synthesis noticeably while $\alpha$-cycfodextrin showed little effect on tryptophan production.

• Journal of the Korean Applied Science and Technology
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• v.29 no.3
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• pp.435-442
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• 2012

The cytochrome P-450 enzymes (CYP 2A6) regulate many endogenous signaling molecules and drugs. Aryl alkynes such as 2-ethynylnaphthalene are important P450 inhibitors which have been extensively studied as medicines or as an effective chemical probes for profiling mouse liver microsomal P-450. Here we have synthesized indole-based novel P450 inhibitor, 5-ethynyl indole 3, and showed that it has successfully inhibited CYP 2A6 by chemical inhibition reaction. By using HPLC equipped with a photo diode array(PDA) detector, all of the peaks derived from the enzymatic reaction have been characterized.

• Biomedical Science Letters
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• v.2 no.1
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• pp.121-126
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• 1996

Specific monoclonal antibodies(mAbs) against bis-indole alkaloids related to vinblastine were established to develop a simple and specific immunoassay system for the quantitation of Vinca alkaloids. Vinca alkaloids were extracted from tissue cultured cells of periwinkle plant (Vinca rosea L.). Spleen cells from Balb/c mice immunized with vinblastine-bovine serum albumin(VBL-BSA) conjugate as immunogen were fused with myeloma cells(Sp2/0-Ag.14) in the presence of polyethylene glycol. In the preliminary experiments, 32 clones which highly reacted with VBL-BSA conjugate were selected by ELISA(Enzyme-linked immunosorbent assay).These clones were further analyzed by inhibition assay of ELISA. The results obtained with two typical monoclonal antibodies, KN-1 and KN-2, were described. KN-1 exhibited considerable reactivities with soluble dimeric bis-indole alkaloids, whereas no cross reacted with monomeric bis-indole alkaloids. However KN-2 showed cross reactivity with mono- and di-meric bis-indole alkaloids. Furthermore, KN-1 was applied to the immunoassay system for determining the VBL amounts of in vitro cultured cell extracts. This assay system could detect dimeric vinca alkaloid as low as 0.05 nM.

• Applied Biological Chemistry
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• v.28 no.1
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• pp.28-35
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• 1985

Potato tubers treated at $4^{\circ}C$ for 4 weeks were irradiated with a dose of 0.12 kGy from $^{60}Co$ source and stored at $20^{\circ}C,\;70{\sim}90%$ humidity for 5 weeks. Changes of ${\alpha}-amylase$, peroxidase, indole acetic acid oxidase, indole acetic acid synthesizing enzyme activities were determined. In addition, treatment of gibberellin or indole acetic acid to tubers irradiated were carried out to examine reversal of sprout-inhibition of tubers irradiated. Results are as follows; 1. Irradiation by ${\gamma}-ray$ at 0. 12 kGy dose inactivated easily the enzyme activities in vitro. $D_{37}$ values obtained were 0.94, 0.36 kGy for ${\alpha}-amylase$ and peroxidase, respectively 2. Complete inhibition of the toter sprouting was resulted by the irradiation of tubers with a dose of 0.12 kGy. 3. The indole acetic acid oxidase activity increased 2 times immediately after irradiation. Meanwhile, indole acetic acid synthesizing activity decreased about $50{\sim}75%$ for 5-week storage in irradiated potatoes, whereas the activity increased about 3.5 times along with sprouting in non-irradiated tubers. 4. In morphological aspects, deformed buds with necrosis in the meristmatic tissue were developed in irradiated tubers. Treatment of gibberellin or indole acetic acid at the concentration of 100 or 20 ppm to the irradiated tubers reversed the sprout-inhibition partially. Nevertheless, the deformed buds remained without change.

• Bulletin of the Korean Chemical Society
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• v.6 no.5
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• pp.291-295
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• 1985

The efficiency of photosensitization of methyl linoleate (ML) oxidation by N-acetyl-L-trypophan(NAT) and 3-methyl indole(scatole) was markedly enhanced by increased concentration of ML in ethanol solution. The fluorescence intensities of sensitizers were observed to be quenched by ML, indicating that ML interacts with the indole excited singlet state. The inhibition of photosensitization by azide demonstrated a possible role of singlet oxygen in the photosensitization. The steady state kinetic treatment of azide inhibition of photosensitization was expected to show linear increase of reciprocal yield of ML oxidation product vs. reciprocal ML concentration at constant azide concentration, but the actual slope was nonlinear. This indicates another competing reaction involved in the photosensitization, As a possible competing reaction, electron transfer from ML to the excited sensitizer was proposed, since the measured fluorescence quenching rate constant closely resembled electron transfer rate constant determined from ML concentration dependence of oxidation product formation.

• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.784-795
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• 2018

Bacillus amyloliquefaciens Pc3 was isolated from Antarctic seawater with antifungal activity. In order to investigate the metabolic regulation mechanism in the biosynthesis of lipopeptides in B. amyloliquefaciens Pc3, GC/MS-based metabolomics was used when exogenous indole was added. The intracellular metabolite profiles showed decreased asparagine, aspartic acid, glutamine, glutamic acid, threonine, valine, isoleucine, hexadecanoic acid, and octadecanoic acid in the indole-treated groups, which were involved in the biosynthesis of lipopeptides. B. amyloliquefaciens Pc3 exhibited a growth promotion, bacterial total protein increase, and lipopeptide biosynthesis inhibition upon the addition of indole. Besides this, real-time PCR analysis further revealed that the transcription of lipopeptide biosynthesis genes ituD, fenA, and srfA-A were downregulated by indole with 22.4-, 21.98-, and 26.0-fold, respectively. It therefore was speculated that as the metabolic flux of most of the amino acids and fatty acids were transferred to the synthesis of proteins and biomass, lipopeptide biosynthesis was weakened owing to the lack of precursor amino acids and fatty acids.

• Archives of Pharmacal Research
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• v.18 no.5
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• pp.351-355
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• 1995

The fecal tryptophanase activities were $0.267{\pm}0.10$ for rats and $0.185{\pm}0.01{\;}{\mu}mole/min/g$ wet feces for humans. The activities of indole pyruvate degradation to indole, indole pyruvate lyase, of these feces were $0.051{\pm}0.02$ and $0.046{\pm}0.01{\;}{\mu}mole/min/g$ wet feces, respectively. The optimal pH values of tryptophanase and indole pyruvate lyase were 5.5-7.5 and 5.5-6.5, respectively. When the intestinal flora or E. coli HGU-3 was cultured in GAM broth having six different pH values (5 to 10), the activities of tryptophanase and indole pyruvate IYilse in the medium adjusted at pH 6 were dramatically induced by elevating the pH to 9. However, when intestinal microflora were inoculated in the medium containing lactulose, the pro¬ductions of these enzymes were dramatically inhibited and the pH of the medium was lower than that of the control.

• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.186-195
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• 2015

Anthranilate synthase (AS) is a key enzyme in the biosynthesis of tryptophan (Trp), which is the precursor of bioactive metabolites like indole-3-acetic acid and other indole alkaloids. Alpha anthranilate synthase 2 (OsASA2) plays a critical role in the feedback inhibition of tryptophan biosynthesis. In this study, two vectors with single (F124V) and double (S126F/L530D) point mutations of the OsASA2 gene for feedback-insensitive ${\alpha}$ subunit of rice anthranilate synthase were constructed and transformed into wildtype Dongjinbyeo by Agrobacterium-mediated transformation. Transgenic single and double mutant lines were selected as a single copy using TaqMan PCR utilized nos gene probe. To select intergenic lines, the flanking sequence of RB or LB was digested with a BfaI enzyme. Four intergenic lines were selected using a flanking sequence tagged (FST) analysis. Expression in rice (Oryza sativa L.) of the transgenes resulted in the accumulation of tryptophan (Trp), indole-3-acetonitrile (IAN), and indole-3-acetic acid (IAA) in leaves and tryptophan content as a free amino acid in seeds also increased up to 30 times relative to the wildtype. Two homozygous event lines, S-TG1 and D-TG1, were selected for characterization of agronomic traits and metabolite profiling of seeds. Differentially expressed genes (DEGs), related to ion transfer and nutrient supply, were upregulated and DEGs related to co-enzymes that work as functional genes were down regulated. These results suggest that two homozygous event lines may prove effective for the breeding of crops with an increased level of free tryptophan content.

• The Korean Journal of Mycology
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• v.41 no.2
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• pp.118-126
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• 2013

The experiment was carried out to analyze the inhibition effect of plant pathogenic fungi and growth promotion activity induced by the bacterial strains isolated from peatmoss. Among the isolated bacterial strains, B10-2, B10-4, B10-5 and B10-6 which showed more than 30% inhibition rate against Botrytis cinerea and Rhizoctonia solani in vitro, were further analyzed in the greenhouse for the growth promotion activity on lettuce (Lactuca sativa), pak-choi (Brassica compestris L. ssp. chinensis) and Chinese cabbage (Brassica campestris L. ssp. pekinensis). The results showed the treatment of B10-4 on lettuce showed the highest growth promotion activity with the leaf area ($169.17cm^2$), fresh weight (leaf: 40.29 g, root: 8.80 g)and dry weight (leaf: 11.24 g, root: 4.17 g), which was about two folds as compared to control. On pak-choi, the growth promotion rate was the highest with the leaf area of $112.87cm^2$, leaf fresh weight of 60.70 g, root fresh weight of 3.37 g, leaf dry weight of 14.34 g, and root dry weight of 1.90 g. As a result of treatment of B10-13 on chinese cabbage, the growth promotion rate was the highest with the leaf area ($293.56cm^2$), fresh weight (leaf: 113.67 g, root: 2.40 g) and dry weight (leaf: 6.03 g, root: 0.53 g). The production of Indole Acetic Acid (IAA) and Indole-3-Butylic Acid (IBA) were also analyzed in these bacterial isolates. The IAA and IBA analyses were carried out in all bacterial isolates each day within the 5 days of incubation period. The highest production of IAA was observed with $112.57{\mu}g/mg$ protein in B10-4 after 3 days of incubation and IBA production was the highest in B10-2 with $58.71{\mu}g/mg$ protein after 2 days of incubation. Also, phosphate solubilizing activity was expressed significantly in B10-13 in comparison to that of other bacterial isolates. Bacterial identification showed that B10-2 was Bacillaceae bacterium and B10-5 was Bacillus cereus, B10-4 and B10-6 were Bacillus sp. and B-13 was Staphylococcus sp. by ITS sequence.