• Journal of Microbiology and Biotechnology
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• v.29 no.1
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• pp.105-113
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• 2019

Although siderophore compounds are mainly biosynthesized as a response to iron deficiency in the environment, they also bind with other metals. A few studies have been conducted on the impact of heavy metals on the siderophore-mediated iron uptake by microbiome. Here, we investigated siderophore production by a variety of rhizosphere fungi under different concentrations of $Zn^{2+}$ ion. These strains were specifically isolated from the rhizosphere of Panax ginseng (Korean ginseng). The siderophore production of isolated fungi was investigated with chrome azurol S (CAS) assay liquid media amended with different concentrations of $Zn^{2+}$ (50 to $250{\mu}g/ml$). The percentage of siderophore units was quantified using the ultra-violet (UV) irradiation method. The results indicated that high concentrations of $Zn^{2+}$ ion increase the production of siderophore in iron-limited cultures. Maximum siderophore production by the fungal strains was detected at $Zn^{2+}$ ion concentration of $150{\mu}g/ml$ except for Mortierella sp., which had the highest siderophore production at $200{\mu}g/ml$. One potent siderophore-producing strain (Penicillium sp. JJHO) was strongly influenced by the presence of $Zn^{2+}$ ions and showed high identity to P. commune (100% using 18S-rRNA sequencing). The purified siderophores of the Penicillium sp. JJHO strain were chemically identified using UV, Fourier-transform infrared spectroscopy (FTIR), and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF-MS) spectra.

• Korean Journal of Soil Science and Fertilizer
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• v.28 no.3
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• pp.287-294
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• 1995

This study demonstrates the influence of environmental conditions, other than iron, on pyoverdin production by fluorescent Pseudomosonas. In slightly acidic pH conditions(pH 6), cell yield was reduced while the siderophore production per cell yield was increased. The optimum temperatures for the siderophore production and cell yield was $19^{\circ}C$ and $28^{\circ}C$ for 7NSK2 and $12^{\circ}C$ and $19^{\circ}C$ for ANP15. The carbon and nitrogen balance showed that at low C : N ratio of the growth medium (higher nitrogen concentration), both cell yield and siderophore production was reduced. Use of different carbon sources revealed that citrate as a carbon source facilitated iron uptake and resulted in a significant reduction in siderophore production. However, at the late exponential phase, the iron content in the cell biomass was not significantly different from those grown in glucose or succinate. From these results it can be suggested that the environmental factors other than iron may also influence siderophore production by fluorescent pseudomonas.

• The Journal of the Korean Society for Microbiology
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• v.34 no.5
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• pp.461-470
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• 1999

Growth under conditions of iron-restriction and the production of siderophore was examined in Vibrio mimicus ATCC 33653. This strain grew and multiplied in the presence of the high-affinity iron chelators ethylenediamine-di (o-hydroxyphenylacetic acid). Chrome azurol S (CAS) agar and solution were used to detect the production of siderophore under these condition. Siderophore could be detected in the iron-restricted culture supernatants. The siderophore was extracted from iron-restricted culture supernatants by phenol-chloroform-ether method and purified by Dowex ion-exchange and Sephadex G-25 gel filtracton chromatography. The purified siderophore was confirmed by paper chromatography and HPLC. The Purified siderophore enhanced the growth of V. mimicus when the bacterium was grown in iron limited medium. Injection of both the siderohore and the bacteria to mice resulted in more rapid death than that of the only bacteria. However, the siderophore did not show lethality to mice and any toxicity to cell line like HeLa and U937.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.415-421
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• 2003

A bacterium, Pseudomonas fluorescens 2112, that is antagonistic against a red-pepper blight-causing fungus, Phytophthora capsici, was isolated from the local soil of Gyongju, Korea. This strain formed an orange-colored clear halo zone on chrome azurol S (CAS) blue agar, suggesting the production of a siderophore in addition to an antifungal antibiotic. The optimal culture conditions for siderophore production by P. fluorescens 2112 were 30-h cultivation at $25^{\circ}C$ and pH 6.5 in King's B medium. The presence of $20{\mu}g/ml\;of\;Fe^3+$ ion or EDDHA promoted the production of siderophore in King's B medium. The siderophore was purified from culture broth by CM-Sephadex C-25 and Sephadex G-25 column chromatographies. The UV spectra of the purified siderophore was the same as that of pyoverdins or pseudobactins. The molecular mass was 1,958 Da determined by FAB-rlass spectrometer, and the amino acid composition analysis showed that the purified siderophore consisted of glycine/threonine/serine/glutamic acid/alanine/lysine with the molar ratio of 3:2:1:1:1:1, DL-Threo-${\beta}$-hydroxyaspartic acid and $N^{\delta}$-hydroxyornithine, two of the essential constituents of pyoverdin, were also found. The purified siderophore pyoverdin showed strong in vitro and in vivo antagonistic activities against phytophthora blight-causing P. capsici. Especially in an in vivo pot test, the siderophore protected red-pepper Capsicum annum L. very well from the attack of P. capsici. These results indicated that the purified siderophore of P. fluorescens 2112 played a critical role in the biocontrol of the red-pepper blight disease, equivalent to treatment by P.fluorescens 2112 cells.

• Korean Journal of Microbiology
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• v.53 no.2
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• pp.97-102
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• 2017

Catechol type siderophore different from 2, 3-dihydroxybenzoic acid (DHB) was produced from Acinetobacter sp. B-W grown in medium containing L-glutamic acid as both carbon and nitrogen sole sources at $28^{\circ}C$. Optimal concentration of glutamic acid for siderophore production was 3% and production of siderophore was decreased above 3% glutamic acid. In previous report, siderophore, 2, 3-DHB was produced from strain B-W grown in medium containing glucose as carbon source and glutamic acid as nitrogen source. Rf value of siderophore produced from strain B-W grown in medium glutamic acid as both carbon and nitrogen sole sources at $28^{\circ}C$ was 0.32, while 2, 3-DHB was 0.84 in butanol-acetic acid-water (12:3:5) as developing solvent. Antioxidative activity of 2, 3-DHB was not detected in that siderophore produced from glutamic acid. Catechol nature of siderophore was detected by Arnow test. Although in iron-limited media optimal cell growth was identified at $36^{\circ}C$, significant quantities of siderophore were produced only at $28^{\circ}C$. Biosynthesis of siderophore was strongly inhibited by growth at $36^{\circ}C$. Production of siderophore was completely inhibited by $10{\mu}M\;FeCl_3$.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.266-271
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• 2018

Effect of plasmid curing of Acinetobacter sp. B-W on the production of siderophore from glutamic acid as both carbon and nitrogen sole sources was investigated. Plasmid cured mutant of strain B-W lost the ability to produce siderophore from glutamic acid at $28^{\circ}C$. Transformant E. coli $DH5{\alpha}$ harboring 20 kb plasmid, that was isolated from wild type of strain B-W produced siderophore from glutamic acid as both carbon and nitrogen sole sources at $28^{\circ}C$, but, not at $36^{\circ}C$. Production of siderophore from glutamic acid by transformant E. coli $DH5{\alpha}$ was completely inhibited by $10{\mu}M\;FeCl_3$. In previous report, catechol nature of siderophore produced from glutamic acid by strain B-W was detected by Arnow test. The siderophore produced from glutamic acid by transformant E. coli $DH5{\alpha}$ was also catechol type. Rf value of siderophore produced from transformant E. coli $DH5{\alpha}$ grown in medium glutamic acid as both carbon and nitrogen sole sources at $28^{\circ}C$ was 0.32 in butanol-acetic acid-water (12:3:5) as developing solvent. Rf value of the siderophore was the same with that of wild type of strain B-W. Thus a single plasmid of 20 kb seemed to be involved in the production of siderophore from glutamic acid.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1500-1512
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• 2017

In this study, the siderophore-producing characteristics and conditions of Bacillus sp. PZ-1 were investigated and the enhancement of siderophores on Pb uptake and translocation in Brassica juncea were determined. Results of single factor experiment showed that glucose, pH, and $Pb(NO_3)_2$ could stimulate PZ-1 growth and siderophore production. The maximum siderophore production of 90.52% siderophore units was obtained by response surface methodology optimization at the glucose concentration of 21.84 g/l, pH 6.18, and $Pb(NO_3)_2$ concentration of $245.04{\mu}mol/l$. The type of siderophore was hydroxamate and its concentration in the fermentation broth amounted to $32.24{\mu}g/ml$. Results of pot experiments indicated that the siderophores enhanced B. juncea to assimilate more Pb from soil with the uptake ratio from 1.04 to 2.74, and to translocate more Pb from underground to overground with the TF values from 1.21 to 1.48. The results revealed that Bacillus sp. PZ-1 could produce abundant siderophores and might be potentially used to augment the phytoextraction of Pb from soil.

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.217-224
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• 2021

Rice blast caused by Pyricularia oryzae, which is a major threat to food security worldwide, markedly decreases the yield of rice. Some rhizobacteria called 'plant growth-promoting rhizobacteria' inhibit plant pathogens and improve plant growth by secreting iron-chelating siderophores. The decreased availability of iron adversely affects the survival of pathogens, especially fungal pathogens, in the rhizosphere. This study aimed to determine the morphological diversity of siderophore-producing bacteria, analyze the type of siderophores produced by the bacteria, and examine their growth-inhibitory activity against Pyricularia oryzae. The rhizobacteria were isolated from the rhizosphere of Sembada Hitam variety of black rice plants in Pakem, Sleman, Yogyakarta, Indonesia. In total, 12 distinct isolates were screened for the production of siderophores. It was found that 9 out of 12 bacteria produced siderophore and most of them were Gram positive bacteria. The best siderophore-producing isolates with different type of siderophore were used in further studies. The IS3 and IS14 isolates were found to be the best siderophore producer that produced hydroxamate and mixed type of hydroxamate-carboxylate type of siderophore, respectively. In the dual culture assay, IS14 showed a strong antagonistic effect against Pyricularia oryzae by the 81.17% inhibition.

• Journal of Environmental Science International
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• v.29 no.9
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• pp.925-933
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• 2020

To develop eco-friendly microbial inoculants, siderophore-producing bacteria were isolated and identified, and their production characteristics and plant growth-promoting abilities were investigated. A strain S21 was isolated from rhizosphere of Korean perilla (Perilla frutescens) and identified as Enterobacter amnigenus by phenotypic properties and 16S rRNA gene sequencing. The highest siderophore production was obtained in a medium containing 0.5% fructose, 0.1% urea, 0.5% K₂HPO₄ and 0.1% succinic acid. By using this improved medium, siderophore production increased by 2.5 times compared to that of basal medium. The strain S21 showed insoluble phosphate solubilizing, ammonification and antifungal activities, and also produced hydrolytic enzymes (protease and lipase), indoleacetic acid and 1-aminocyclopropane-1-carboxylate deaminase. Our data suggest that E. amnigenus S21 is a potential candidate that can be used as eco-friendly biocontrol agent and biofertilizer.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.258-263
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• 2008

Late blight, one of the most important disease in many agricultural crops, is caused by Phytophthora infestans. Fusarium wilt is a vascular disease of many plants caused by Fusarium oxysporum. Some bacteria isolated from rhizosphere were screened for their ability to inhibit the growth of F. oxysporum and P. infestans. Productions of siderophore, $\beta-1$,3-glucanase, hydrogen cyanide and chitinase by 4 isolated strains were examined. Among them, Bacillus sp. RFO41 most effectively inhibited the growth of F. oxysporum. The highest productions of siderophore and $\beta-l$,3-glucanase were shown in the culture of Bacillus sp. RFO41. Bacillus strain PS2 was most effective against P. infestans. PS2 showed the highest production of chitinase and hydrogen cyanide. A significant relationship was shown between the antagonistic effects of isolates against F. oxysporum and P. infestans and their production level of siderophore, $\beta-1$,3-glucanase, hydrogen cyanide, and chitinase.