• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.156-162
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• 1988

Optimal conditions for the formation and regeneration of protoplasts of the cellulolytic fungus Penicillium verruculosum were investigated. Among the various commercial cell wall lytic enzymes tested, 0.5%(w/ v) Novozym 234 was the most effective for protoplast formation. The highest yield of protoplast exceeding 4.5$\times$10$^6$/m$\ell$ obtained when 400mg of 20 hr-old mycelia was incubated with 0.5%(w/v) Novozym 234 at 3$0^{\circ}C$ for 1 hr. The best osmotic stabilizer for the isolation and re-generation of protoplasts was 0.7M sorbital (pH 5.6) and 0.6M MgSO$_4$(pH 5.6), respectively. When 0.6M MgSO$_4$was added as osmotic stabilizer to the complete medium, the maximum regeneration frequency obtained was 4.6-27.8%. Micromorphological change of giant protoplasts into hyphae was observed during incubation in the regeneration liquid medium.

• Journal of Microbiology and Biotechnology
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• v.18 no.11
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• pp.1741-1746
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• 2008

This study was conducted to select endophytic actinomycetes as biocontrol agents against Chinese cabbage clubroot caused by Plasmodiophora brassicae. A total of 81 endophytic actinomycetes were isolated from surface-sterilized roots of Chinese cabbage that was grown on paddy field and upland soils collected from various locations in Korea. By using 16S ribosomal DNA (rDNA) gene sequencing, they were classified to 8 actinobacterial genera. The genus Microbispora (67%) was most frequently isolated, followed by Streptomyces (12%) and Micromonospora (11%). Three of the 81 isolates, when inoculated in germinated Chinese cabbage seeds and then transplanted to pots, effectively suppressed the occurrence of a post-inoculated strain of P. brassicae in the pots. They showed control values of 58% for strain A004, 33% for strain A011, and 42% for strain A018. Based on cell wall components, morphological characteristics, and phylogenetic analyses, the three antagonistic isolates were identified as Microbispora rosea subsp. rosea (A004 and A011) and Streptomyces olivochromogenes (A018). Further researches on the field efficacy and action modes of the three actinomycetes are in progress.

• Applied Biological Chemistry
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• v.43 no.3
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• pp.169-173
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• 2000

For the development of enzyme detergent capable of effectively washing at low temperature, a bacterium producing alkaline protease was isolated from soil samples, and properties of the enzyme were investigated. The selected strain was Gram negative, rod shape$(0.6{\sim}0.7{\times}1.3{\sim}2.6\;{\mu}m\;in\;size)$ and motile. It had the degradation activity of aesculin, gelatin and casein, and was catalase-positive. The cell wall components was meso-DAP, and G+C mole contents was 43.3%. From these results, the strain was identified as Acinetobacter sp. KN-27. The activity of alkaline protease by this strain peaked with 3,300 D.U/mL after 36 hours in the liquid culture at $40^{\circ}C$. The optimal pH and temperature of the enzyme were pH 9 and $60^{\circ}C$, respectively. Alkaline protease produced by Acinetobacter sp. KN-27 has shown two active bands on the electrophoresis of native gel.

• Journal of the Earthquake Engineering Society of Korea
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• v.26 no.1
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• pp.13-22
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• 2022

This study proposes a low-cycle fatigue life derived from measurement points on pipe elbows, which are components that are vulnerable to seismic load in the interface piping systems of nuclear power plants that use seismic isolation systems. In order to quantitatively define limit states regarding leakage, i.e., actual failure caused by low-cycle fatigue, in-plane cyclic loading tests were performed using a sine wave of constant amplitude. The test specimens consisted of SCH40 6-inch carbon steel pipe elbows and straight pipes, and an image processing method was used to measure the nonlinear behavior of the test specimens. The leakage lines caused by low-cycle fatigue and the low-cycle fatigue curves were compared and analyzed using the relationship between the relative deformation angles, which were measured based on each of the measurement points on the straight pipe, and the moment, which was measured at the center of the pipe elbow. Damage indices based on the combination of ductility and dissipation energy at each measurement point were used to quantitatively express the time at which leakage occurs due to through-wall cracking in the pipe elbow.

• Journal of Microbiology and Biotechnology
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• v.33 no.7
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• pp.895-908
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• 2023

Disposal of waste containing heavy metals into the environment is a major threat to human health and can result in toxic or chronic poisoning in aquatic life. In the current study, metal-resistant Raoultella ornithinolytica was isolated from metal-contaminated samples collected from the Tanjaro River, located southwest of Sulaymaniyah, Iraq. R. ornithinolytica was identified by partial amplification of 16S rRNA. The uptake potency of heavy metals was assessed using inductively coupled plasma-optical emission spectroscopy (ICP-OES) and indicated that R. ornithinolytica removed 67, 89, 63.4, 55.6, 56.5, 65, and 61.9% of Cd, Pb, Cr, Ni, Zn, Co, and Fe, respectively. These removal rates were influenced by temperature, pH, and contact time; at 35℃ and pH 5 with a change in the incubation time, the reduction rate improved from 89 to 95% for Pb, from 36.4 to 45% for Cu, and from 55.6 to 64% for Ni. Gene analysis indicated that R. ornithinolytica contained pbrT, chrB, nccA, iroN, and czcA genes, but the pcoD gene was absent. Energy-dispersive X-ray spectroscopy (EDS) images showed evidence of metal ion binding on the cell wall surface with different rates of binding. Transmission electron microscopy (TEM) detected different mechanisms for metal particle localization; cell surface adsorption was the main mechanism for Pb, Zn, and Co uptake, while Cd, Ni, and Fe were accumulated inside the cell. The current study describes, for the first time, the isolation of R. ornithinolytica from metal-contaminated water, which can be used as an eco-friendly biological expedient for the remediation and detoxification of metals from contaminated environments.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.6
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• pp.879-886
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• 2016

As is in all economic activities, the highest yield per unit area is the main goal in animal production, while addressing the temperamental needs of animals often is ignored. Animal welfare is not only an ethical fact; it also has an economic value. Spatial environmental enrichment contributes positively to animal welfare by addressing their behavioral and mental requirements. The present study was conducted to determine the effects of weaning and spatial environmental arrangements on behaviors of goat-kids. Experimental groups were arranged in structured and unstructured spatial environments. Roughage feeder, semi-automatic concentrate feeder, bunk, bridge, and wood block were placed in the structured environment. No equipment was placed in the unstructured environment and paddock sides were enclosed with an iron sheet to prevent bipedal stance and to provide environmental isolation. In the study 10 male and 10 female Turkish Saanen goat kids were used in each group. Spatial environmental arrangements did not have significant impacts on the growth performance of kids (p>0.05). All objects in the structured group were accepted by the kids. Average use ratios of roughage feeder, semi-automatic concentrate feeder, bunk, bridge and wood block were observed as 19.3%, 14.0%, 12.6%, 3.8%, and 0.7%, respectively. There were significant differences between before- and after-weaning in use of all objects except for underneath bridge ($p{\leq}0.05$). Concentrate feed consumption, locomotion, and resting behaviors in kids showed significant differences by structural group and growth period. Roughage consumption was similar between groups, while it differed by growth period ($p{\leq}0.05$). Interaction frequency was significantly higher in structured group (p = 0.0023). Playing behavior significantly differentiated based on the growth period rather than on groups ($p{\leq}0.05$). Playing behavior significantly decreased after weaning. Abnormal oral activity was significantly higher in the structured group before weaning ($p{\leq}0.05$). Despite there being no installations facilitating climbing and bipedal stance, the kids of the unstructured group were able to exhibit 1/3 as much bipedal stance behavior as the kids of the structured group through leaning over slippery paddock wall or over their groupmates. Bipedal stance behavior of unstructured group was similar before and after weaning, while bipedal stance behavior before weaning was about 2 times that of after weaning in structured group. It was concluded that unstructured environmental arrangement limited the behavior repertoire of the goat kids.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.6
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• pp.1002-1011
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• 2020

Objective: This study was conducted to determine the composition and diversity of the fungal flora at various control points in cheese ripening rooms of 10 dairy farms from six different provinces in the Republic of Korea. Methods: Floor, wall, cheese board, room air, cheese rind and core were sampled from cheese ripening rooms of ten different dairy farms. The molds were enumerated using YM petrifilm, while isolation was done on yeast extract glucose chloramphenicol agar plates. Morphologically distinct isolates were identified using sequencing of internal transcribed spacer region. Results: The fungal counts in 8 out of 10 dairy farms were out of acceptable range, as per hazard analysis critical control point regulation. A total of 986 fungal isolates identified and assigned to the phyla Ascomycota (14 genera) and Basidiomycota (3 genera). Of these Penicillium, Aspergillus, and Cladosporium were the most diverse and predominant. The cheese ripening rooms was overrepresented in 9 farms by Penicillium (76%), while Aspergillus in a single farm. Among 39 species, the prominent members were Penicillium commune, P. oxalicum, P. echinulatum, and Aspergillus versicolor. Most of the mold species detected on surfaces were the same found in the indoor air of cheese ripening rooms. Conclusion: The environment of cheese ripening rooms persuades a favourable niche for mold growth. The fungal diversity in the dairy farms were greatly influenced by several factors (exterior atmosphere, working personnel etc.,) and their proportion varied from one to another. Proper management of hygienic and production practices and air filtration system would be effective to eradicate contamination in cheese processing industries.

• Microbiology and Biotechnology Letters
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• v.33 no.4
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• pp.295-301
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• 2005

About seven hundred bacterial strains were collected from Jeot-Kal, a Korean traditional fermented fishes, in various Korean districts. One of the strains designated JKK238 has its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The JKK238 strain was isolated from Oh-Jeot, a kind of fermented shrimps, of Kangkyeung in Korea, and was identified as Bacillus subtilis based on its physiological characteristics, fatty acids compositions of cellular wall, and 16S rDNA sequence analysis. We isolated simply antimicrobial lipopeptides (AMLP) by $25\%$ ammonium sulfate precipitation of 3 days-old tryptic soy broth cultures of the JKK238 strain. Further analysis of AMLP revealed that B. subtilis JKK238 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families simultaneously. Above results indicate that the JKK238 strain can be added to the limited number B. subtilis strains reported to co-produce the three kinds of lipopeptide families.

• Journal of Mushroom
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• v.13 no.3
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• pp.270-273
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• 2015

Hypsizigus marmoreus is commercially the most important edible mushroom in Japan. This mushroom is usually cultivated for a longer period (about 85~120 days) than other mushroom. In order to develop a new cultivar that has a shortened cultivation period, the genome analysis of this strain has been considered. This study aims to obtain parental monokaryotic strains reproducing 'Haemi' cultivar in Hypsizigus marmoreus for reference genome sequencing. The mycelia were cultured in MCM and MYG media for various incubation periods. Homogenized mycelia were treated with commercial cell wall degrading enzymes to maximize protoplasts production yield from Hypsizigus marmoreus. The greatest number of protoplasts was obtained from mycelia cultured in MCM media for 3 days using Novozyme enzyme. The isolated protoplasts were grown in regeneration agar media after two weeks. Regenerated colonies were picked and moved on separated dishes for microscopic observation. Neohaplonts regenerated from dikayotic strains were identified by the absence of clamp connections. We confirmed that one of monokaryotic strains is a parental strain by crossing with an original compatible strain of 'Haemi' cultivar. This parental strain will be used for reference genome sequence analysis.

• Korean Journal of Plant Tissue Culture
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• v.28 no.3
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• pp.141-146
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• 2001

Protoplasts were isolated from cotyledons, hypocotyls, and mesophyll tissues of three species of Capsicum species (C. anuumm, C. bacatuum, and C. chacoense). Combination of Cellulysin (1%) and Macero-zyme (0.25%) in 0.65 M sorbitol was found to be the most effective for the digestion of cell wall, regardless of the Capsicum species. Antioxidant MES (2-［N-Morpholino］ethanesulfonic acid) in the enzyme solution helped protoplasts overcome browning. After 5 days of initial culture, Cell division occurred in modified K8p medium containing 1~5 mg/L zeatin, 0.5 mg/L IAA, 0.1~0.5 mg/L TDZ, and 1 mg/L 2,4-D under continuous dark condition at $25^{\circ}C$. Semi-solid agarose culture method was more effective than liquid culture, and it also protected the cells from browning caused by polyphenolic compound released during protoplast culture. A total of 4000 calli were obtained from protoplast culture of different capsicum species. All of these calli were transferred to the 100 combinations of regeneration media using various plant growth regulators; TDZ, IAA, 2ip, BAP, NAA, and zeatin. These calli derived from protoplast of three species of capsicum were, however, not differentiated into shoots.