• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.493-500
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• 2021

Argyrins are a group of anticancer and antibacterial octapeptide bioactive substances isolated from myxobacteria. In this study, we showed that the myxobacterium Archangium gephyra MEHO_001, isolated in Korea, produces argyrins A and B. MEHO_001 cells tend to aggregate when cultured in liquid media. Hence, a dispersion mutant, MEHO_002, was isolated from MEHO_001. The MEHO_002 strain produced approximately 3.5 times more argyrins than that produced by the wild-type strain MEHO_001. We determined the whole-genome sequence of A. gephyra MEHO_002 and identified a putative argyrin biosynthetic gene cluster comprising five genes, arg1-arg5, encoding non-ribosomal peptide synthases and tailoring enzymes. Inactivation of arg2 by plasmid insertion disrupted argyrin production. The amino acid sequences of the proteins encoded by arg2-arg5 of A. gephyra MEHO_002 were 90-98% similar to those encoded by the argyrin biosynthetic genes of Cystobacter sp. SBCb004, an argyrin-producing myxobacterium with identical domain organization.

• Microbiology and Biotechnology Letters
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• v.33 no.1
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• pp.41-50
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• 2005

An organic acid tolerance mutant (M-200) was obtained from Leuconostoc mesenteroides KCCM 35471, followed by the screening procedure using a specific organic acid medium (lactic acid: acetic acid, 2:1). The characteristics of the acid tolerance M-200 and the wild type LM-W were examined at various temperature and pH ranges $(l0-30^{\circ}C$ of temp, 3.5-4.5 of pH). The growth of strain M-200 at HCl adjusted medium $(10^{\circ}C\;and\;pH 3.5)$ was observed. In the case of organic acid adjusted medium, the strain showed its growth at the pH range of 3.8. When the strain M-200 was used as a starter for Kimchi fermentation, a constant acid level (0.55) was observed during the whole fermentation period. This result indicates that the strain produces a proper level of acid content for the Kimchi fermentation. This result also indicates that the edible period of Kimchi can be extended to 3.5 fold compare to the result obtained from the LM-W used Kimchi fermentation. However the excess use of the strain M-200 showed the inhibition of growth of Lactobacillus plantarum, low lactic acid level content and low level of organoleptic test. In the case of organic acid content during the Kimchi fermentation, the strain M-200 showed relatively low production rate compare to the wild type (M-200: 3.5 mg/L at 21 days of fermentation, LM-W: 7 mg/L at 21 days of fermentation). Therefore a mixed Kimchi starter containing M-200 and other strains probably maintain a good Kimchi quality during the fermentation.

• Journal of Microbiology and Biotechnology
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• v.27 no.3
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• pp.633-643
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• 2017

To examine the pro-apoptotic role of the human ortholog (YPEL5) of the Drosophila Yippee protein, the cell viability of Saccharomyces cerevisiae mutant strain with deleted MOH1, the yeast ortholog, was compared with that of the wild-type (WT)-MOH1 strain after exposure to different apoptogenic stimulants, including UV irradiation, methyl methanesulfonate (MMS), camptothecin (CPT), heat shock, and hyperosmotic shock. The $moh1{\Delta}$ mutant exhibited enhanced cell viability compared with the WT-MOH1 strain when treated with lethal UV irradiation, 1.8 mM MMS, $100{\mu}M$ CPT, heat shock at $50^{\circ}C$, or 1.2 M KCl. At the same time, the level of Moh1 protein was commonly up-regulated in the WT-MOH1 strain as was that of Ynk1 protein, which is known as a marker for DNA damage. Although the enhanced UV resistance of the $moh1{\Delta}$ mutant largely disappeared following transformation with the yeast MOH1 gene or one of the human YPEL1-YPEL5 genes, the transformant bearing pYES2-YPEL5 was more sensitive to lethal UV irradiation and its UV sensitivity was similar to that of the WT-MOH1 strain. Under these conditions, the UV irradiation-induced apoptotic events, such as FITC-Annexin V stainability, mitochondrial membrane potential (${\Delta}{\psi}m$) loss, and metacaspase activation, occurred to a much lesser extent in the $moh1{\Delta}$ mutant compared with the WT-MOH1 strain and the mutant strain bearing pYES2-MOH1 or pYES2-YPEL5. These results demonstrate the functional conservation between yeast Moh1 and human YPEL5, and their involvement in mitochondria-dependent apoptosis induced by DNA damage.

• The Korean Journal of Mycology
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• v.49 no.3
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• pp.337-349
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• 2021

This study aimed to isolate wild yeasts from water and soil sample of the Nonsan stream and Sapgyoho (lake) in Chungcheonnam-do, Korea, and to further characterize previously unrecorded wild yeast strains. In total, 102 strains, representing 55 different species of wild yeast were isolated from 95 samples collected from the Jangseoncheon and Ipchoncheon of Nonsan stream in Jellabuk-do and Chungcheonnam-do. Among these, 33 strains were isolated from alkalophilic yeast extract-peptone-dextrose (YPD) medium (pH 9.0), and 9 strains were isolated concurrently on general YPD medium (pH 6.5) and alkalophilic medium. Seventeen strains of Cryptococcus laurentii were predominantly isolated. Additionally, 65 strains, representing 27 different species of wild yeast were isolated from 58 samples obtained from Sapgyoho (lake). Among the 82 isolated wild yeast strains, 8 strains, including Candida fructus JSC 72-1(JSL-GGU 015), had not previously been recorded. All 8 previously unrecorded yeasts were oval in shape except C. fructus JSC72-1(JSL-GGU-015), and only the Filobasidium chernovii JSC39-1(JSL-GGU-013) strain formed spores. All strains except Pseudosydowia eucalypti JSC23-6(JSL-GGU-012) grew well in yeast extract-peptone-dextrose (YPD) and yeast extract-malt extract media and grew in vitamin-free medium. Four strains, including P.eucalypti JSC23-6(JSL-GGU-012) grew well in 15% NaCl-containing YPD medium. F.chernovii JSC39-1(JSL-GGU-013) and Sirobasidium intermedium JSC7-3(JSL-GGU-014) assimilated lactose, and five strains, including F. chernovii JSC39-1(JSL-GGU-013) also assimilated starch. All strains were resistant to 800 ppm of Ca, Cu, Li, and Mg ions.

• Korean Journal of Microbiology
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• v.38 no.3
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• pp.221-229
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• 2002

Japanese encephalitis virus (JEV) persistence was established and maintained in Vero cell culture for over 1 year. Eleven clones of plaque morphology mutant JEV, with large and small plaque sizes, were obtained from the cell culture supernatant. Genomic RNA replication efficiency of the mutants in naive Vero cell appeared to correspond to their different plaque sizes. No significant changes in envelop protein ORF or in non-coding regions at both ends of the RNA genome suggested that there could be an unidentified factor(s) playing role in JEV attenuation. Unlike to the replication of wild-type JEV, the mutants did not induce severe degree of cytopathic effect in Vero cells upon infection. While obvious decrease of Bcl-2 and its mRNA expression and sharp increase of p53 in naive Vero cells infected with either wild-type JEV or the large plaque-forming mutant, those changes were not observed with the small plaque-forming one. Together with these observation, internucleosomal DNA fragmentation and chromosomal DNA profile in the Vero cells infected with the mutants suggest that an overall changes in cytopathic effect in the plaque morphology mutants-infected cells should be primarily due to the reduced genomic RNA replication and the compromised degree of p53-independent apoptosis by the virus infection at least in part.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.207-217
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• 1998

Total of 78 strains were characterized based on the morphological characteristics of colonies isolated on Schroth, and New & Kerr's media for selection of hypervirulent wild-type Agrobacterium spp from galls, hairy root-like process and soil of Populus, Malus, Salix and Diopyros in Korea. Among them, 48 strains were able to induce tumors in carrot disc. Hypervirulent A. tumefaciens SP101 and SM042 were identified as biotype 1 and biotype 2, respectively, These strains formed fast growing, larger tumors as compared to those induced by other strains. The binary vector pGA643 with kanamycin resistant gene was mobilized from E. coli MC100 into A. tumefaciens strain SM042 isolated from soil, and/or disarmed vector PC2760 using a triparental mating method with E. coli HB101/pRK2013, and transconjugants, A. tumefaciens SM643 and PC643 were obtained in minimal media containing kanamycin and tetracycline. Tobacco tissues were cocultivated with conjugant Agrobacterium and then transferred to selective medium with 2,4-D and kanamycin to induce the transformants. Calli were formed more efficiently in cocultivation with A. tumefaciens SM643 than that with A. tumefaciens PC643. Most of calli transformed with A. tumefaciens PC643 were friable and regenerated into normal plantlets, while the calli transformed with A. tumefaciens SM643 were compact, hard, and mixed with friable calli. The friable calli formed normal shoots, while compact calli did not form shoots but only grew to typical compact tumor calli. When the shoots formed directly from tobacco stems without callus induction after transformation by A. tumefaciens SM643 with wild-type Ti-plasmid, normal transformed plants can be induced without using disarmed Ti-plasmid.

• Journal of Life Science
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• v.22 no.3
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• pp.417-427
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• 2012

We compared the transcriptome in response to propanol stress in wild-type and propanol-resistant mutant Escherichia coli using the DNA microarray technique. The correlation value of RNA expression between the propanol-treated wild type and the untreated-one was about 0.949, and 50 genes were differentially expressed by more than twofold in both samples. The correlation value of RNA expression between the propanol-treated mutant and the untreated one was about 0.951, and 71 genes in two samples showed differential expression patterns. However, the values between the wild type and mutant, regardless of propanol addition, were 0.974-0.992 and only 1-2 genes were differentially expressed in the two strains. The representative characteristics among differentially expressed genes in W3110 or P19 treated with propanol compared to untreated samples were up-regulation of hest shock response genes and down-regulation of genes relating to ribosome biosynthesis. In addition, many genes were regulated by transcription regulation factors such as ArcA, CRP, FNR, H-NS, GatR, or PurR and overexpressed by sigma factor RpoH. We confirmed that RpoH mediated an important host defense function in propanol stress in E. coli W3110 and P19 by comparison of cell growth rate among the wild type, rpoH disruptant mutant, and rpoH-complemented strain.

• Journal of Microbiology and Biotechnology
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• v.25 no.5
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• pp.672-680
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• 2015

As a global regulatory gene in Streptomyces, afsR can activate the biosynthesis of many secondary metabolites. The effect of afsR on the biosynthesis of a phenazine metabolite, lomofungin, was studied in Streptomyces lomondensis S015. There was a 2.5-fold increase of lomofungin production in the afsR-overexpressing strain of S. lomondensis S015 N1 compared with the wild-type strain. Meanwhile, the transcription levels of afsR and two important genes involved in the biosynthesis of lomofungin (i.e., phzC and phzE) were significantly upregulated in S. lomondensis S015 N1. The optimization of metal chlorides was investigated to further increase the production of lomofungin in the afsR-overexpressing strain. The addition of different metal chlorides to S. lomondensis S015 N1 cultivations showed that CaCl₂, FeCl₂, and MnCl₂ led to an increase in lomofungin biosynthesis. The optimum concentrations of these metal chlorides were obtained using response surface methodology. CaCl₂ (0.04 mM), FeCl₂ (0.33 mM), and MnCl₂ (0.38 mM) gave a maximum lomofungin production titer of 318.0 ± 10.7 mg/l, which was a 4.1-fold increase compared with that of S. lomondensis S015 N1 without the addition of a metal chloride. This work demonstrates that the biosynthesis of phenazine metabolites can be induced by afsR. The results also indicate that metal chlorides addition might be a simple and useful strategy for improving the production of other phenazine metabolites in Streptomyces.

• The Plant Pathology Journal
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• v.25 no.1
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• pp.77-85
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• 2009

Pseudomonas fluorescens strain Gpf01, isolated from ginseng rhizosphere showed antiviral activity against Cucumber mosaic virus, when tested in a local host of CMV, Chenopodium amaranticolor. Transposon mutant library of Gpf01 was prepared using pGS9::Tn5 and the mutant Gpf01-RS19 was found to loose antiviral production. We developed primers from the flanking region of Tn5 and found a cosmid clone pAV1123, harboring 1.2 kb antiviral compound producing (avcf01) locus. When a sub-clone pPH9, which carried 9.3 kb region of pAV1123, was introduced into antivirus deficient P. fluorescens wild type strain B16, it exhibited antiviral activity. Using Tn3-gus mutagenesis and complementation analysis, it was found that the genes related to antiviral activity production resided in a 9.3 kb HindIII-HindIII fragment of pAV1123, indicating that the plasmid carries an essential genes promoting antiviral activity.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.33-37
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• 2010

Rhizobacterial strain isolated from barren soil, Bacillus subtilis RFO41 exhibits a high level of phosphate solubilizing activity and produces some phytohormones. Its promoting effect on the growth of Xanthium italicum Moore, a wild plant growing at lakeside barren land and thus a good candidate plant for revegetation of barren lakeside was evaluated in the in situ test for 19 weeks at Lake Paro, Kangwon-do. Strain RFO41 could enhance the dry weight of X. italicum by 67.7%. It also increased the shoot length of X. italicum plant by 21.1% compared to that of uninoculated control. Both growth enhancements had statistical significance. However, the inoculation did not show any effect on the root growth, which might be due to the breakage of tiny root. Denaturing gradient gel electrophoresis analysis showed that the inoculated bacteria were maintained in the soils, and the indigenous bacterial community did not exhibit any significant change. This plant growth promoting capability may be utilized as an environment-friendly and low cost revegetation method, especially for the sensitive areas such as barren lakeside lands.