• Korean Journal of Microbiology
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• v.39 no.3
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• pp.128-134
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• 2003

Transformation-associated recombination (TAR) cloning is based on co-penetration into yeast spheroplasts of genomic DNA along with TAR vector DNA that contains 5'- and 3'-sequences (hooks) specific for a gene of interest, followed by recombination between the vector and the human genomic DNA to establish a circular YAC. Typically, the frequency of recombinant insert capture is 0.01-1% for single-copy genes by TAR cloning. To further refine the TAR cloning technology, we determined the effect of GC content on target hooks required for gene isolation utilizing the $Tg\cdot\AC$ mouse transgene as the targeted region. For this purpose, a set of vectors containing a B1 repeated hook and Tg AC-specific hooks of variable GC content (from 18 to 45%) was constructed and checked for efficiency of transgene isolation by radial TAR cloning. Efficiency of cloning decreased approximately 2-fold when the TAR vector contained a hook with a GC content ～${\leq}23$% versus ～40%. Thus, the optimal GC content of hook sequences required for gene isolation by TAR is approximately 40%. We also analyzed how the distribution of high GC content (65%) within the hook affects gene capture, but no dramatic differences for gene capturing were observed.

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.316-323
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• 1992

Studies on the changes in rheological properties, molecular weight distribution and dextran yield after being reacted in lO%(w/w) sucrose concentration were performed with crude dextransucrase produced from Leuconostoc mesenteroides isolated from Sikhae. The reaction rate of dextran formation was monitored by sugar analysis with HPLC and by the changes in apparent viscosity. According to the periodate oxidation test, the dextran produced in this experiment was estimated to have 89% $\alpha$-(1->6) main linkages and 11% $\alpha$-(1->) side linkages. The rheological properties of the dextran solution formed changed with reaction time, and it was related to the changes in molecular weight distribution of dextran as determined by GPC analysis. As the reaction proceeded, the rheological behavior changed from Newtonian to non-Newtonian, showing Binghampseudoplastic and thixothropic flow behavior. The apparent viscosity of dextran formed solution increased with increasing reaction time, reached a maximum value of 2680 cP ($\gamma$=$33.75s^{-1}$, $25^{\circ}C$) by enzyme reaction for 8 hours, and then decreased. The temperature dependency of dextran formed solutions was well expressed by the Arrhenius equation and the activation energy reached a maximum value of 1.69 kcal/mole by enzyme reaction for 8 hours.

• Korean Journal of Microbiology
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• v.53 no.1
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• pp.29-38
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• 2017

In this study, we investigated the phylogenetic diversity of the bacterial community and isolation of extremely halophilic bacteria using culturomics in a gray solar saltern. The number of bacterial living cells, enumerated in a gray solar saltern by direct fluorescence microscopy was three to four orders of magnitude greater than those enumerated by plate counts, suggesting the distribution of 'viable but non-culturable bacteria'. The biodiversity of bacterial communities in a gray solar saltern was investigated by pyrosequencing, 1,778 OTUs of bacteria were comprised of 18 phyla 46 classes 85 orders 140 families 243 genera with 6.16 diversity index. Archaea communities were composed of 3 phyla 6 classes 7 orders 7 families 38 genera with 4.95 diversity index from 643 OTUs. Totally 137 isolates were isolated by 59 different cultural methods based on culturomics considering culture media and conditions suitable for the growth of extremely halophilic bacteria. Phylogenetic analyses of extremely halophilic isolates based on 16S rRNA gene sequences, extremely halophilic isolates were composed of 4 phyla and 11 genera. Haloterrigena and Haloferax can be successfully isolated from culturomics. These culturomics were effective methods for collection of diversity of extremely halophilic bacteria.

• Korean Journal of Microbiology
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• v.34 no.4
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• pp.219-224
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• 1998

The cyclic metabolism of poly(3-hydroxyhutyrate-co-3-hydroxyvalerate) synthesized from glucose by Alcaligenes sp. SH-69 in the presence or absence of new carbon substrate was investigated. In batch culture, the content and weight average molecular weight of the copolymer already stored in the cell decreased rapidly when there was no other carbon source available. After the depletion of carbon source, the amount of high molecular weight copolymer decreased more rapidly than that of low molecular weight copolymer, and as a result, average molecular weight distribution shifted to the lower value. The addition of a mixture of glucose and levulinic acid when the initial carbon substrate, glucose, was nearly depleted supported the continual increase in cell mass and the accumulation of poly(3HB-co-3HV) with high molar fraction of 3HV. However, solvent fractionation of the polymer with acetone revealed the degradation of pre-existing polyhydroxyalkanoale (PHA) in parallel with the synthesis of PHA from new carbon substrate. Even though PHAs obtained from each substrate alone were the copolymer of 3HB and 3HV, it was found that the polymer accumulated in the cells grown by sequential feeding was mainly physical mixture of two poly(3HB-co-3HV) copolymers containing different molar fractions of 3HV.

• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.323-331
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• 2019

White jelly fungus (Tremella fuciformis Berk; TF) has been used as a traditional medicine in Asia; it is known to prevent hypertension, aging, cancer, and arteriosclerosis. This study aimed to investigate the anti-diabetic effects of fermented Tremella fuciformis Berk (FTF) ethanol extracts fermented with L. rhamnosus BHN-LAB 76. We show that FTF increases the ${\alpha}$-glucosidase inhibitory activity and suppress the adipogenesis of 3T3-L1 adipocytes. These inhibitory effects of FTF are accompanied by the regulation of the phosphorylation of AMPK, JNK, and Akt. These data demonstrate that FTF not only inhibits adipogenesis by affecting the adipogenic signaling, but also increases the anti-diabetic effects by regulating the insulin signaling pathway. Therefore, we suggest that the FTF can be used for developing functional food and cosmetics materials.

• The Korean Journal of Mycology
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• v.27 no.1 s.88
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• pp.10-14
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• 1999

A single phase composter was constructed by modifying the conventional mixer of sawdust for the cultivation of oyster mushroom Pleurotus ostreatus. The machine was designed on the basis of 3-phase-1 system which was controlled in prewetting, pasteurization and fermentation processes. In composting 200 kg of straw and cotton waste in the machine, it took 20 minutes in prewetting step and also to hours at $65^{\circ}C$ in pasteurization process. Postfermentation by aerothermophiles was completed by treating the compost at $45^{\circ}C-50^{\circ}C$ for 48 hours which was shorten 24 hours from the conventional method. In the postfermentation at high temperature, forced aeration and/or vigorous mixing process(es) played a great role in the improvement of spawn quality. The growth of mycelium of oyster mushroom was excellent in the culture combinated with 3 parts of surface inoculation and 7 parts of mechanical mixing.

• Microbiology and Biotechnology Letters
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• v.13 no.4
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• pp.409-415
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• 1985

This study has been carried out in order to investigate the bacterial quality of fish meat paste products and the characteristics of isolated thermodurics from the products. Twenty samples of crab-flavored fish stick (Kematsal), 23 samples of plate fish meat paste (Panomuk, Kamaboko), 5 samples of fried fish meat paste (Tigimomuk), 2 samples of roasted fish meat paste (Puduromuk, Chikuwa), 20 samples of fish sausage were collected from processing plants and supermarkets in Pusan, Korea during the period from May to October in 1984. The results obtained are as follows. Amont the samples collected from supermarkets, roasted fish meat paste and fried fish meat paste marked hish counts in coliforms and fungi while very low in the samples of crab-flavored fish stick and plate fish meat paste. Salmonella was not detected in all the samples examined and Staphylococcus aureus was detected only in fried fish meat paste, Thermoduric bacteria were detected less than 10$^2$/g in the samples of crab-flavored fish stick and plate fish meat paste, which might come from subsidiary materials such as starch and seasonings. Among the isolated bacteria, distribution of the proteolytics were more than 87％ and the lipolytics were less than 20％. Gram positive bacteria was more than 70％ in crab-flavored fish stick and plate fish meat paste, 47.3％ in fried fish meat paste. And rod in shape was almost more than 90％ in all the samples. The most heat resistant bacterium isolated from the samples was identified as a Bacillus licheniformis(named B. licheniformis CR-11). The strain showed strong proteolytic activity and also grew well at above 2$0^{\circ}C$. The growth rate and generation time of CR-11 strain were 0.31 hr$^{-1}$ , 2.24 hr at 2$0^{\circ}C$, 0.64 hr$^{-1}$ , 1.09 hr at 3$0^{\circ}C$ and 0.78 hr$^{-1}$ , 0.89 hr at 35$^{\circ}C$. Heat resistance value of the spores of CR-11 strain suspended in phosphate buffer solution was D$_{85}$ $^{\circ}C$=41.9 min, D$_{90}$ $^{\circ}C$=27.9 min, D$_{95}$ $^{\circ}C$=10.2 min, D$_{100}$ $^{\circ}C$=4.3 min (Z=13.8$^{\circ}C$)

• The Korean Journal of Mycology
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• v.13 no.1
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• pp.41-47
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• 1985

Eighteen plant residues were added to soil and the amended soil was inoculated with chlamydospores (5,000 cells/g soil) of Fusarium solani causing root rot of ginseng in soil to test the effects of amendment using pea (Pisium sativa L.) as an index plant. Distributional conditions of microogranisms in soil were compared with each other before and after dealing with soil amendment by plant debries. Infection rate by index plant's infectivity showed a higher degree in the treatment of wheat crushed than in control group, and followed by stalk of sweet potato, chinese cabbage, ginseng leaves and soybean pod. On the other hand, the de­creasing order of infection rate was root of garlic, welsh onion, cabbage leaf and stalk, green onion stalk, wheat straw and barley straw. In comparison with control group, the propagules of fungi increased in the treatment of ginseng leaves, soybean ground, wheat crushed, maize stalk, and chinese cabbage, but decreased in the root of garlic, cabbage, and barley straw. Population of total bacteria increased in the treatment of soybean ground, chinese cabbage, radish stalk, welsh onion, and wheat crushed, but decreased in barley straw, tobaco root, ginseng stalk, and wheat straw. The numbers of actinomycetes increased only in the treatment of soybean ground in a comparison with control and also decreased in the garlic stalk and tobaco root. The propagules of Fusarium spp increased in the treatment of chinese cabbage, welsh onion, radish stalk, wheat crushed, and sweet potato stalk, wheat crushed, and sweet potato stalk, but decreased in the treat­ment of wheat straw, ginseng leaves, and cabbage than control.

• Korean Journal of Microbiology
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• v.45 no.2
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• pp.222-227
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• 2009

The seasonal variation of bacterial populations in the Korean cattle (Hanwoo) feedlots dispersed in the 20 cities of Gyeongsangbuk-do, Korea, was monitored for two years (2006 and 2007) to provide quantitative criteria for good agricultural management. Outside the feedlots, the average falling bacterial populations of the air were $3{\pm}1$ CFU/$cm^2\cdot15$ min (n=63) in a year. Inside the feedlots, the average falling bacterial populations of the air were $8{\pm}1$ CFU/$cm^2\cdot15$ min (n=63) in the spring, $16{\pm}2$ CFU/$cm^2\cdot15$ min (n=69) in the summer, $7{\pm}2$ CFU/$cm^2\cdot15$ min (n=69) in the autumn, and $6{\pm}1$ CFU/$cm^2\cdot15$ min (n=70) in the winter. Without using the summer data, the average falling bacterial population of the air was $7{\pm}1$ CFU/$cm^2\cdot15$ min, which was not statistically significant (P=0.37). The average bacterial populations in the cattle drinking water of the cattle feedlots were $4,710{\pm}780$ CFU/ml (n=65) in the spring, $10,430{\pm}1170$ CFU/ml (n=65) in the summer, $4,820{\pm}700$ CFU/ml (n=64) in the autumn, and $2,510{\pm}530$ CFU/ml (n=64) in the winter. Without using the summer data, the average bacterial population of the drinking water was $4,000{\pm}400$ CFU/ml, which was statistically significant (P=0.027). The average frequency of Escherichia coli O157 inside the feedlots was 5% (n=65) in the spring, 72% (n=65) in the summer, 67% (n=66) in the autumn, and 29% (n=66) in the winter on the basis of soil samples of the year 2007. The results indicate that most of the Escherichia coli O157 strains distributed in the summer and autumn was disappeared in the spring through the cold weather of the winter.

• Journal of Food Hygiene and Safety
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• v.35 no.5
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• pp.477-488
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• 2020

Recently, foodborne illness outbreaks linked to fresh produce are being increasingly reported in the United States, the EU, and Korea as well. Some of this increase may be due to improved surveillance, increase in consumption, change in consumers' habits, and complex distribution systems. Garlic chive is a green, fresh-cut vegetable consumed year-round as a nutrition-rich herb in Korea. It is also prone to contamination with foodborne pathogens during pre-harvest, as amendment with high amounts of livestock manure or compost to soil is required in its cultivation. Our aim in this study was to evaluate microbial contamination of garlic chives, garlic chives cultivation soil, compost, and irrigation water in the southern part of Korea. Samples were collected in A, B, and C regions in 2019 and 2020, and 69, 72, 27, and 40 of garlic chives, soil, compost, and irrigated water, respectively, were analyzed for the presence of sanitary indicator bacteria (total aerobic bacteria, coliforms and Escherichia coli), Bacillus cereus, Staphylococcus aureus, pathogenic E. coli, E. coli O157:H7, Listeria monocytogenes, and Salmonella spp. In A, B, and C regions, levels of total aerobic bacteria, coliform, B. cereus, and S. aureus on all samples were between 1.14 and 8.83 log CFU/g, 0.43 and 5.01 log CFU/g, 0.41 and 5.55 log CFU/g, and 1.81 and 6.27 log CFU/g, respectively. B. cereus isolated from garlic chives and environmental samples showed β-hemolysis activity. Incidence of S. aureus in garlic chive and its production environments in 2020 was different from 2019. In this study, B. cereus and S. aureus were the only pathogenic microorganisms detected in all samples. As a result, this work suggests that continuous monitoring in the production and pre-harvest environment is required to improve hthe hygiene and safety of garlic chive.