• The Plant Pathology Journal
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• v.33 no.3
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• pp.276-287
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• 2017

RcsA is a positive activator of extracellular polysaccharide (EPS) synthesis in the Enterobacteriaceae. The rcsA gene of the soft rot pathogen Pantoea sp. strain PPE7 in Pleurotus eryngii was cloned by PCR amplification, and its role in EPS synthesis and virulence was investigated. The RcsA protein contains 3 highly conserved domains, and the C-terminal end of the open reading frame shared significant amino acid homology to the helix-turn-helix DNA binding motif of bacterial activator proteins. The inactivation of rcsA by insertional mutagenesis created mutants that had decreased production of EPS compared to the wild-type strain and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii. The Pantoea sp. strain PPE7 rcsA gene was shown to strongly affect the formation of the disease symptoms of a mushroom pathogen and to act as the virulence factor to cause soft rot disease in P. eryngii.

• Food Science of Animal Resources
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• v.41 no.6
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• pp.967-982
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• 2021

Probiotics are living microorganisms that, when administered in adequate amounts, provide a health benefit to the host and are considered safe. Most probiotic strains that are beneficial to human health are included in the "Lactic acid bacteria" (LAB) group. The positive effects of probiotic bacteria on the host's health are species-specific and even strain-specific. Therefore, evaluating the probiotic potential of both wild and novel strains is essential. In this study, the probiotic characteristics of Lactobacillus brevis KT38-3 were determined. The strain identification was achieved by 16S rRNA sequencing. API-ZYM test kits were used to determine the enzymatic capacity of the strain. L. brevis KT38-3 was able to survive in conditions with a broad pH range (pH 2-7), range of bile salts (0.3%-1%) and conditions that simulated gastric juice and intestinal juice. The percentage of autoaggregation (59.4%), coaggregation with E. coli O157:H7 (37.4%) and hydrophobicity were determined to be 51.1%, 47.4%, and 52.7%, respectively. L. brevis KT38-3 produced β-galactosidase enzymes and was able ferment lactose. In addition, this strain was capable of producing antimicrobial peptides against the bacteria tested, including methicillin and/or vancomycin-resistant bacteria. The cell-free supernatants of the strain had high antioxidant activities (DPPH: 54.9% and ABTS: 48.7%). Therefore, considering these many essential in vitro probiotic properties, L. brevis KT38-3 has the potential to be used as a probiotic supplement. Supporting these findings with in vivo experiments to evaluate the potential health benefits will be the subject of our future work.

• Journal of Microbiology and Biotechnology
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• v.14 no.6
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• pp.1211-1216
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• 2004

A MNNG (N-methyl-N'-nitro-N-nitrosoguanidine) induced chromium resistant strain ($Cr^{r}18$) of unicellular cyanobacterium Anacystis nidulans has been isolated and characterized. The resistant strain could grow (although restricted to $50\%$ of control) in chromium concentration (180${\mu}M$) lethal to the wild-type. Sublethal ($160{\mu}M$) concentration of $Cr^{6+}$ significantly reduced (13-$37.5$) all the photosynthetic pigments of A. nidulans with maximum reduction in phycoerythrin followed by ChI $\alpha$. Pigments of A. nidulans were drastically decreased in lethal concentration of Cr^{6+} with maximum reduction in phycoerythrin ($75\%$) and allophycocyanin ($67.5\%$). Resistant strain $Cr^{r}18$ resisted toxic effects of sublethal and lethal concentrations of $Cr^{6+}$ on photosynthetic pigments as revealed by less decrease in pigments as compared to A. nidulans. Effect of $Cr^{6+}$ stress was also studied on nitrogen assimilation and phosphate uptake. Sublethal concentration of $Cr^{6+}$ drastically reduced ($71.5\%$) nitrate uptake by A. nidulans while a decrease of $29\%$ was observed in strain $Cr^{r}18$. Short (2 day) exposure of A. nidulans and its resistant strain $Cr^{r}18\;to\;Cr^{6+}$ did not affect nitrate reductase and glutamine synthetase (transferase), whereas longer (10 day) exposure to $Cr^{6+}$ lowered activities of both enzymes in A. nidulans but not significantly in the strain $Cr^{r}18$. Ammonium uptake by both strains was not affected by $Cr^{6+}$. Thus, $Cr^{6+}$ affected photosynthetic pigments, nitrogen assimilation, and phosphate uptake of A. nidulans, while strain $Cr^{r}18$ was able to resist toxic effects of the metal. Advantages of using strain $Cr^{r}18$ for bioremediation purposes have been evaluated by studying $Cr^{6+}$ removal from the solution. Resistant strain $Cr^{r}18$ was able to remove $33\%$ more $Cr^{6+}$ than A. nidulans and thus it can prove to be a good candidate for bioremediation of $Cr^{6+}$ from polluted waters.

• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.648-657
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• 2008

In this study, we used functional genomic strategies, proteomics and quantitative real-time (qRT)-PCR, to advance our understanding of genes associated with fumonisin production in the fungus Fusarium verticillioides. Earlier studies have demonstrated that deletion of the FCC1 gene, which encodes a C-type cyclin, leads to a drastic reduction in fumonisin production and conidiation in the mutant strain (FT536). The premise of our research was that comparative analysis of F. verticillioides wild-type and FT536 proteomes will reveal putative proteins, and ultimately corresponding genes, that are important for fumonisin biosynthesis. We isolated proteins that were significantly upregulated in either the wild type or FT536 via two-dimensional polyacrylamide gel electrophoresis, and subsequently obtained sequences by mass spectrometry. Homologs of identified proteins, e.g., carboxypeptidase, laccase, and nitrogen metabolite repression protein, are known to have functions involved in fungal secondary metabolism and development. We also identified gene sequences corresponding to the selected proteins and investigated their transcriptional profiles via quantitative real-time (qRT)-PCR in order to identify genes that show concomitant expression patterns during fumonisin biosynthesis. These genes can be selected as targets for functional analysis to further verify their roles in $FB_1$ biosynthesis.

• Interdisciplinary Bio Central
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• v.3 no.1
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• pp.4.1-4.7
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• 2011

Forty-seven wild accessions of Lotus japonicus Regal (Japanese trefoil) indigenous to Japan were investigated for nine morphological characters. Average temperature and annual precipitation were negatively correlated with stem color and seed weight. On the other hand, latitude was positively correlated with these traits. Consequently, accessions from sites at higher latitudes with low temperatures and precipitation tend to have dark red stems and heavy seeds. Cluster analysis based on nine morphological characters classified 47 wild accessions into six major groups. Cluster I included four accessions of tall and erect plants. These plants are phenotypically similar to commercial variety 'Empire'. Cluster II consisted of three accessions of creep plants with pale red stems. Cluster III contained 24 accessions that had average values for all morphological characters evaluated. Cluster IV included two accessions of erect plants with rounded leaflets and dark red stems. Cluster V included four accessions of small, creep plants with pale red stems. Cluster VI included seven accessions of small and erect plants, a phenotype that also applies to ?Gifu B-129?, which is used as experimental strain worldwide. These data were deposited into LegumeBase, an online database (http://www.legumebase.brc.miyazaki-u.ac.jp/) supported by the National BioResource Project (NBRP) in Japan.

• Journal of Microbiology and Biotechnology
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• v.21 no.12
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• pp.1228-1235
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• 2011

We compared the gene expression among four clinical and five environmental V. vulnificus isolates, using a cDNA microarray containing 131 genes possibly associated with pathogenicity, transport, signal transduction, and gene regulations in the pathogen. cDNAs from total RNAs of these isolates were hybridized into the cDNA microarray using the cDNA of the wild-type strain MO6-24/O as a reference. We focused on selecting differentially expressed (DE) genes between clinical and environmental isolates using a modified t-statistic. We could detect two statistically significant DE genes between virulent isolates and less-virulent isolates with a marginal statistical significance (p-value of 0.008). These were genes putatively encoding pilin and adenlyate cylase. Real time-PCR confirmed that these two selected genes transcribed in significantly higher levels in virulent isolates than in less-virulent isolates. Mutants with lesions in the gene encoding pilin showed significantly higher $LD_{50}$ values than that of wild type.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1519-1528
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• 2017

Foodborne contamination and salmonellosis caused by Salmonella Enteritidis (S. Enteritidis) are a significant threat to human health and poultry enterprises. Outer membrane vesicles (OMVs), which are naturally secreted by gram-negative bacteria, could be a good vaccine option because they have many biologically active substances, including lipopolysaccharides (LPS), outer membrane proteins (OMPs), and phospholipids, as well as periplasmic components. In the present study, we purified OMVs derived from S. Enteritidis and analyzed their characteristics through silver staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis. In total, 108 proteins were identified in S. Enteritidis OMVs through liquid chromatography tandem mass spectrometry analysis, and OMPs, periplasmic proteins, and extracellular proteins (49.9% of total proteins) were found to be enriched in the OMVs compared with bacterial cells. Furthermore, native OMVs used in immunizations by either the intranasal route or the intraperitoneal route could elicit significant humoral and mucosal immune responses and provide strong protective efficiency against a lethal dose (~100-fold $LD_{50}$) of the wild-type S. Enteritidis infection. These results indicated that S. Enteritidis OMVs might be an ideal vaccine strategy for preventing S. Enteritidis diseases.

• KSBB Journal
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• v.27 no.3
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• pp.172-176
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• 2012

In this study, microalgae Arthrospira platensis (A. platensis) mutants induced by ethyl methane sulfonate (EMS) and further selection for resistance of cerulenin, a potent inhibitor of fatty acid synthase, were characterized. The mutants selected by $2{\mu}M$, $5{\mu}M$ and $10{\mu}M$ of cerulenin were designated EC2, EC5 and EC10, respectively. Under normal growth conditions, the mutants and parental strain exhibited similar growth pattern. The mutants of A. platensis showed enhanced lipid accumulation and phycobiliproteins (phycoerythrin, phycocyanin). The lipid content of mutants EC2 and EC5 was about 4.4 and 4.8-fold higher than wild type. The phycoerythrin and phycocyanin content of mutants EC2 and EC5 was increased about 1.5 and 6.9-fold and 1.4 and 3.8-fold, respectively, compared to the wild type. The chlorophyll and carotenoid content of mutants was slightly increased. The high lipid and pigment contents exhibited by A. platensis mutants would make an excellent candidate for the production of commercially interesting biologically active compounds.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.78-89
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• 2000

Protein secretion in filamentous fungi has been shown to be restricted to actively growing hyphal tips. To determine whether an increase in the amount of growing surface area of a fungus can lead to an increase in the amount of protein secretion, we examined secretion in a temperature-sensitive Neurospora crassa mcb mutant that shows a loss of growth polarity when incubated at restrictive-temperature. Incubation of the mcb mutant at restrictive-temperature results in a three- to five-fold increase in the level of extracellular protein and a 20- fold increase in carboxymethyl cellulase activity relative to a wild-type strain. A mutation in the cr-l gene has been shown previously to suppress the apolar growth phenotype of the mcb mutant, and we find that the level of extracellular protein produced by a mcb; cr-l double mutant was reduced to that of the wild-type control. Immunolocalization of a secreted endoglucanase revealed that proteins are secreted mainly at hyphal tips in hyphae exhibiting polar growth and over the entire surface area of bulbous regions of hyphae that are produced following a shift of the mcb mutant to restrictive-temperature. These results support the hypothesis that secretion of extracellular protein by a filamentous fungus can be significantly increased by mutations that alter growth polarity.

• Korean Journal of Microbiology
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• v.26 no.4
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• pp.291-297
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• 1988

Three alleles of rbsB gene, rbsB, rbsB103, and rbsB106 from the wild type, the mutant and the revertant strain, respectively, were cloned for overproduction of proteins under the control of lambda $P_{L}$ promoter. Five different species of precursor and mature ribose-binding proteins were purified to homogeneity using DEAE-Sephadex column chromatography, osmotic shock pocedure, CM-Sephadex column chromatography, and Chromatofocusing column chromaography. pI of the precursor proteins and mature proteins were determined and found to be pH 8.0 and 7.5, respectively. The purified proteins were subjected to amino acid sequencing. The results confirmed the amino acid changes deduced from the DNA sequencing.