• Korean Journal of Microbiology
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• v.37 no.2
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• pp.120-123
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• 2001

Sphingomonas chungbukensis DJ77 is able to use phenanthrene and biphenyl as the sole carbon and energy source. Mitomycin C curing experiment suggested that polyaromatic hydrocarbon (PAH) utilization in strain DJ77 was plasmid-encoded. The plasmid cured strains were failed to grow on the minimal medium sprayed with biphenyl or phenanthrene. This was evident from southern hybridizations using a previously cloned DNA segment as a probe. There were positive signals in the palsmid DNA of the wild-type strain DJ77 and the absence of hybridizations with chromosomal DNA from the plasmid DNA of the wild-type strain DJ77 and the absence of hybridizations with chromosomal DNA from the palsmid-cured mutant strains.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.6
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• pp.506-513
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• 2004

Spirulina produces $\gamma$-linolenic acid (GLA), an important pharmaceutical substance, in a relatively low level compared with fungi and plants, prompting more research to improve its GLA yield. In this study, metabolic flux analysis was applied to determine the cellular metabolic flux distributions in the GLA synthetic pathways of two Spiru/ina strains, wild type BP and a high­GLA producing mutant Z19/2. Simplified pathways involving the GLA synthesis of S. platensis formulated comprise of photosynthesis, gluconeogenesis, the pentose phosphate pathway, the anaplerotic pathway, the tricarboxylic cycle, the GLA synthesis pathway, and the biomass syn­thesis pathway. A stoichiometric model reflecting these pathways contains 17 intermediates and 22 reactions. Three fluxes - the bicarbonate (C-source) uptake rate, the specific growth rate, and the GLA synthesis rate - were measured and the remaining fluxes were calculated using lin­ear optimization. The calculation showed that the flux through the reaction converting acetyl­CoA into malonyl-CoA in the mutant strain was nearly three times higher than that in the wild­type strain. This finding implies that this reaction is rate controlling. This suggestion was sup­ported by experiments, in which the stimulating factors for this reaction $(NADPH\;and\;MgCl_{2})$ were added into the culture medium, resulting in an increased GLA-synthesis rate in the wild type strain.

• International Journal of Industrial Entomology and Biomaterials
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• v.33 no.1
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• pp.15-23
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• 2016

One major constraint in the development and implementation of a successful and cost-effective area-wide integrated pest management (AW-IPM) programme with a SIT component for Bactrocera oleae (Diptera: Tephritidae) is the ability to produce a large number of high quality mass-reared individuals. The aim of this study was to develop a more efficient and practical egg collection system in an attempt to improve the mass-rearing of this species. The following basic parameters were examined: egg production per female, egg hatch, pupal recovery, pupal weight, adult emergence and percentage of fliers. Three different strains (Israel wild-type, France wild-type, and Greece laboratory) were tested and each strain was evaluated for six generations. Female flies of the Israel strain produced significantly more eggs per female than the other two strains, but egg hatch was significantly lower. Egg hatch of the France wild type and the Greece laboratory strain was similar. For all other parameters, there was no significant difference between strains; however, there was a significant generational effect for all parameters observed. As a result of this study, a protocol was developed for the mass-rearing of this species that included the use of large adult holding cages that could house up to 96,000 flies per cage. The newly developed method of egg collection using a flat wax panel as one of the sides of an adult holding cage proved to be cost-effective, efficient, making colony growth easier for industrial mass-rearing.

• KSBB Journal
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• v.21 no.2
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• pp.90-95
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• 2006

Escherichia coli harboring pAC-LYCO4 and pDdxs was used for lycopene production. Three wild type strains of E. coli OW1, MG1655, and W3110 were compared with DH5${\alpha}$ used before for lycopene production. Lycopene productivity of E. coli MG1655 was similar to DH5${\alpha}$ and the highest among those wild type strain. Therefore, MG1655 strain was used for random transposon and NTG mutagenesis to increase lycopene productivity. Through transposon mutation, five transposon mutants with increased lycopene productivity were obtained. It was found that genes knocked out by transposon insertion were treB in Tn1 mutant, B2436 in Tn2 mutant, and rfaH in Tn3, 4, and 5 mutants. Lycopene productivity was the highest in Tn4 mutant among the Tn mutants, which was 6-fold and 8-fold higher in lycopene concentration and content, respectively, in comparison with those obtained with wild type strain. NTG4 mutant was acquired with NTG mutation. The highest lycopene productivity of 6 mg/L and 4 mg/g DCW was obtained from the NTG4 mutant when arabinose of 0.013 mM was added for induction of dxs, rate-limiting gene of MEP pathway. The lycopene productivity of NTG4 mutant was increased 18-fold and 12-fold in lycopene concentration and content, respectively when comparing with the wild type strain.

• Journal of Life Science
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• v.34 no.2
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• pp.79-85
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• 2024

SlyA is known as a transcriptional regulator that regulates the expression of hemolysin (HlyE) in E. coli, a member of the Enterobacteriaceae family such as Salmonella. However, Salmonella has the slyA gene but lacks the hlyE gene. Then, because we were curious about the role of SlyA in Salmonella, we constructed and explored a mutant strain with a deletion of the slyA gene. S. Typhimurium CK295 (ΔslyA) was constructed using an allelic exchange approach. In a comparative analysis between the wild-type and the CK295 strain, no significant differences were observed in growth characteristics, motility, total protein analyses, and secreted protein analyses. However, the CK295 strain exhibited slightly reduced biofilm formation compared to the wild-type. Interestingly, as a result of comparing the survival ability in macrophages, the mutant strain showed a 60% decrease in survival ability compared to the wild-type. To evaluate toxicity in mice, mortality was measured after oral administration to 6-week-old BALB/c mice. As a result, the LD₅₀ value of the CK295 (ΔslyA) was more than 100 times higher than that of wild-type S. Typhimurium 𝜒3339 in BALB/c. In conclusion, SlyA is presumed to regulate the expression of genes encoding virulence factors involved in the in vivo survival of Salmonella.

• Journal of Microbiology and Biotechnology
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• v.26 no.4
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• pp.700-709
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• 2016

The mitogen-activated protein kinase HOG1 (high-osmolarity glycerol response pathway) plays a crucial role in the response of yeast to hyperosmotic shock. Trichosporonoides oedocephalis produces large amounts of polyols (e.g., erythritol and glycerol) in a culture medium. However, the effects of HOG1 gene knockout and environmental stress on the production of these polyols have not yet been studied. In this study, a To-HOG1 null mutation was constructed in T. oedocephalis using the loxP-Kan-loxP/Cre system as replacement of the targeted genes, and the resultant mutants showed much smaller colonies than the wild-type controls. Interestingly, compared with the wild-type strains, the results of shake-flask culture showed that To-HOG1 null mutation increased erythritol production by 1.44-fold while decreasing glycerol production by 71.23%. In addition, this study investigated the effects of citric acid stress on the T. oedocephalis HOG1 null mutants and the wild-type strain. When the supplementation of citric acid in the fermentation medium was controlled at 0.3% (w/v), the concentration of erythritol produced from the wild-type and To-HOG1 knockout mutant strains improved by 18.21% and 21.65%, respectively.

• Korean Journal of Veterinary Research
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• v.53 no.2
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• pp.95-101
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• 2013

The study was carried out to evaluate and compare the immune responses in mice experimentally infected with either wild-type or isogenic mutants of Salmonella enterica serovars Enteritidis (SE), Salmonella Typhimurium (ST) and Gallinarum (SG). The mutant strains were constructed by allelic replacement of some virulence-associated genes in the wild-type strains. Seven-week-old female BALB/c mice were orally or intraperitoneally inoculated by injecting bacterial suspension. To evaluate the immune responses, enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunospot (ELISPOT) assay were conducted with serum and fecal samples. As a result, the mice group infected orally with the SE mutant strain showed the highest level of specific IgA-secreting splenocytes, compared to the other groups. The peritoneally injected groups showed the greater levels of IgG1 than the orally injected groups, which was in a good agreement with the previous studies. In addition, the mutant infected groups had the similar secretion levels of antibodies with the wild-type infected groups. These results demonstrated that the SE mutant strain elicited humoral immune response as much as wild-type, implying that it can be useful as a delivery vehicle as well as a candidate of a live attenuated vaccine.

• Journal of Microbiology and Biotechnology
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• v.20 no.12
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• pp.1702-1710
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• 2010

Korean traditional rice wines yakju and takju are generally brewed with nuruk as the source of the saccharogenic enzymes by natural fermentation. To improve the quality of Korean rice wine, the microorganisms in the nuruk need to be studied. The objective of this research was to improve the quality of Korean wine with the wild-type yeast strains isolated from the fermentation starter, nuruk. Only strain YA-6 showed high activity in 20% ethanol. Precipitation of Y89-5-3 was similar to that of very flocculent yeast (>80%) at 75.95%. Using 18S rRNA sequencing, all 10 strains were identified as Saccharomyces cerevisiae. Volatile compounds present in yakju were analyzed by gas chromatography-mass selective detector. The principal component analysis (PCA) of the volatile compounds grouped long-chain esters on the right side of the first principal component, PC1; these compounds were found in yakju that was made with strains YA-6, Y89-5-3, Y89-5-2, Y90-9, and Y89-1-1. On the other side of PC1 were short-chain esters; these compounds were found in wines that were brewed with strains Y183-2, Y268-3, Y54-3, Y98-4, and Y88-4. Overall, the results indicated that using different wild-type yeast strains in the fermentation process significantly affects the chemical characteristics of the glutinous rice wine.

• Journal of Mushroom
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• v.12 no.3
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• pp.181-186
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• 2014

This research was carried out to analyze the genetic variation of 18 wild strain, 2 breed varieties and 20 mutants of Hypsizygus marmoreus by random amplification of polymorphic DNA(RAPD). Also, 3 strains of Lyophyllum decartes and 1 strain of Lyophyllum shimeji were used. These mushrooms were collected from korea, china, Taiwan and Japan. Spores of H. marmoreus JV-2 strain were irradiated by gamma ray for mutagenesis. 40 kind of primers were used for this reaserch. Number of reaction primer were 31. Electrophorectic patterns of RAPD showed genetic variation. In phylogenetic tree, they were divided into seven group. Discriminative differences were observed between wild strain and mutants in H. marmoreus. These results might suggest that these primers and gamma ray irradiation of spores were useful tools for developing new strain for mushroom.

• Journal of Microbiology and Biotechnology
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• v.26 no.8
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• pp.1452-1456
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• 2016

Overexpression of the genes encoding phosphoeneolpyruvate carboxykinase (pckA) and NAD-dependent malic enzyme (maeA) facilitates higher intracellular ATP and NAD(P)H concentrations, respectively, under aerobic conditions in Escherichia coli. To verify a hypothesis that higher intracellular energy reserves might contribute to H₂ fermentation, wild-type E. coli strains overexpressing pckA and maeA were cultured under anaerobic conditions in a glucose minimal medium. Overexpression of pckA and maeA enabled E. coli to produce 3-times and 4-times greater H₂ (193 and 284 nmol, respectively) than the wild type (66 nmol H₂). The pckA and maeA genes were further overexpressed in a hydrogenase-3-enhanced E. coli strain. The hydrogenase-3-enhanced strain (W3110+fhlA) produced 322 nmol H₂, whereas the ATP-enhanced strain (W3110+fhlA+pckA) produced 50% increased H₂ (443 nmol). Total H₂ in the NAD(P)H-enhanced strain (W3110+fhlA+maeA) was similar to that in the control strain at 319 nmol H₂. Possible explanations for the contribution of the increased cellular energy reserves to the enhanced hydrogen fermentation observed are discussed based on the viewpoint of metabolic engineering strategy.