• Korean Journal of Soil Science and Fertilizer
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• v.38 no.1
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• pp.8-14
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• 2005

Mass culturing of two beneficial organisms used as biofertilizers for crops would reduce the risks in production and minimize the capital involved and this demands appropriate media that supports both organism and also selection of organisms that are not antagonistic to each other. A study was initiated to culture a nitrogen fixer (Rhizobium) and phosphate solubilizer (Bacillus megaterium) in a single medium and to study their growth patterns and shelf life in carrier. The growth of Rhizobium and Bacillus megaterium was assessed in different media and a slight modification in the traditional yeast extract mannitol media promoted the growth of both the organisms. The growth of the individual organisms in the modified medium was assessed by estimating the population at regular intervals and compared to their original medium. Maximum population of Rhizobium and phosphobacteria was at 60 hr when the phosphiobacteria inoculation of later was after 48 hr of Rhizobium inoculation. The shelf life of the individual inoculants in the inoculant containing both the organism in a sterile carrier base revealed no significant differences compared to individual organisms inoculated in a sterilized carrier. The population of both organisms in carrier based mixed inoculant remained at $10^8$ cells till 90 days.

• Korean Journal of Environmental Biology
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• v.25 no.1
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• pp.34-41
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• 2007

A study on the distribution of V. parahaemolyticus among sea water, sea mud, and marine products in Hwabuk, Samyang, Daepo, Jungmun, Pyoson, Anduk, Aewol, and Gwakji on the coastal area of Jeju island was conducted from January to December in 2002. The 2,880 total specimens of 960 sea waters, 960 sea mud, 960 marine products were collected and studied for the rate of isolation of V. parahaemolyticus, and biochemical, serological and antibiotic sensitivity tests were performed. A total of 417 strains of V. parahaemolyticus were isolated and identified from 2,880 total specimens. In the test of biochemical properties, 100 of V. parahaemolyticus isolates in the presence of 0.85% NaCl were positive in the utilization of lysine, ornithine, indole, glucose, and mannitol, and negative in the utilization of ONPG, arginine, sodium citrate, urea, tryptophane, inositol, sorbitol, rhamnose, sucrose, and melibiose, $H_2S$ production and VP reaction, while positive or negative in gelatin liquefaction and utilization of amygdalin or arabinose. The isolation rates to the specimen were 161 strains (16.8%) from 960 of sea waters, 137 strains (14.3%) from 960 of sea mud, and 119 strains (12.4%) from 960 of marine products. The isolation rates of V. parahaemolyticus from 8 coastal areas were 14.4% (52/360) in Hwabuk area, 15.3% (55/360) in Samyang area, 13.6% (49/360) in Daepo area, 18.3% (66/360) Jungmun area, 13.1% (47/360) in Pyosun area, 16.4% (59/360) in Anduk area, 12.5% (45/360) in Aewol area and 12.2% (44/360) in Gwakji area, respectively. The distribution of 417 V. parahaemolyticus, isolates was high at Jungmun with 18.3% (66/360), and from sea water with 16.8% (161/960).

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.212-219
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• 2016

Forty-nine pigments were extracted from the collections of 106 pigment producing bacteria from the plant rhizosphere soil. Antibacterial activity test was performed in the subjects of the extracted pigments with plant pathogenic bacteria including Xanthomonas axonopodis and Xanthomonas campestris, and with plant pathogenic fungi including Botrytis cinerea, Colletotrichum acutatum, and Fusarium oxysporum. The yellow pigment by Chryseobacterium sp. RBR9 and the red pigment by of Methylobacterium sp. RI13 showed the antibacterial activities against Xanthomonas axonopodis and Xanthomonas campestris. The violet pigment by Collimonas sp. DEC-B5 showed the antibacterial activity as well as the antifungal activities against Botrytis cinerea and Fusarium oxysporum. Especially, the violet pigment inhibited the growth of Botrytis cinerea more than 65% at MIC $20{\mu}M$. Upon the HPLC analysis result for the isolation of pigment with antifungal activity, violacein (91.6%) and deoxyviolacein (8.4%) were isolated for the pigment by Collimonas sp. DEC-B5. The production amount of the pigment was increased more than 10 times higher when D-mannitol 1.5% and yeast extract 0.2% were added as the nitrogen source to SCB medium. This study suggests that produced violacein by Collimonas sp. DEC-B5 will be effective to control strawberry gray-mold rot fungi by its preventive activity.

• Journal of Mushroom
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• v.22 no.3
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• pp.87-91
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• 2024

Recently, active research in Korea and worldwide has begun to focus on gene function and cultivar development using gene editing tools. This research, in addition to studies on edible mushroom, aims to enhance the physical and biochemical characteristics of mushrooms for applications in materials and substance production. For these studies, efficient isolation of protoplasts from the target mushroom is critical. However, several commercial cell wall-lysing enzyme cocktails, including Novozyme234, Glucanex, and Lysing enzymes, have recently been discontinued. In this study, we aimed to identify alternative enzyme systems to replace the discontinued cell wall-lysing enzymes for stable isolation of protoplasts from Ganoderma lucidum. To select an optimal osmotic buffer, enzyme function in 0.6 and 1.2 M Sorbitol, Sucrose, Mannitol, and KCl was assessed. The effect of reaction time was also evaluated. Protoplast isolation efficiency of each alternative enzyme was tested using lysing enzymes from Trichoderma harzianum, Chimax-N, and Yatalase, either individually or in combination. This matrix of studies identified enzymes and optimal conditions that could replace the discontinued lysing enzymes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.1
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• pp.41-48
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• 2000

A bacterial strain, designated as JK-43, producing extracellular cyclodextrin glucanotransferase (CGTase)[EC 2.4.1.19] was isolated from kimchi. The CGTase from isolated strain JK-43 showed the transglucosylation activity from soluble starch to L-ascorbic acid(AA) compared to those obtained from other strains. A main product formed by this reaction was identified as $2-O-{\alpha}-glucopyranosyl$ L-ascorbic acid(AA-2G) by testing its susceptibility to ${\alpha}-glucosidase$ hydrolysis, the HPLC profiles, and through the elementary analysis. the ${\beta}-CD,\;{\gamma}-CD$, potato starch and corn starch were identified to be suitable glucosyl donor for transglucosylation reaction on AA by CGTase. Acceptor specificity on AA-2G production was examined by use of AA, Iso-AA and AA-2P. Transglucosylation was observed toward AA-2P as well as AA and Iso-AA. The microorganism isolated from kimchi was identified as a strain of Bacillus sp. JK-43 based on the morphological, cultural, biochemical characteristics and partial 16SrDNA sequence analysis. The maximal CGTase production was observed in a medium containing 1.0% soluble starch, 1.0% yeast extract, 1.0% $Na_2CO_3\;0.1%\;K_2HPO_4,\;and\;0.02%\;MgSO_4{\cdot}7H_2O$ with initial pH 7.0. The strain was cultured at $37^{\circ}C$ for 26 hrs with reciprocal shaking.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.5
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• pp.648-656
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• 2008

The objective of this study was to establish conditions for transfection of a foreign gene into somatic cells using cationic lipid reagents and to evaluate the effects of transfection on in vitro development of somatic cell nuclear transfer (SCNT) embryos. Green fluorescent protein (GFP) gene was used as a foreign gene and a non-transfected somatic cell was utilized as a control karyoplast. Monolayers of porcine cells were established and subsequently transfected with a GFP-expressing gene (pEGFP-N1) using three types of transfection reagents (LipofectAMINE PLUS, FuGENE 6 or ExGen500). Donor cells used for SCNT included transfected fetal or adult fibroblasts and oviduct epithelial cells, either serum-fed or serum-starved. Oocytes matured in vitro for 42 h were reconstructed with either transfected or non-transfected porcine somatic cells by electric fusion and activation using a single DC pulse of 1.8 kV/cm for $30{\mu}s$ in $Ca^{2+}$ and $Mg^{2+}-containing$ 0.26 M mannitol solution. Reconstructed oocytes were subsequently cultured in NCSU-23 medium for 168 h and the developmental competence and cell number in blastocyst were compared. There were no significant differences (P>0.05) in fusion, cleavage rates or development to the blastocyst stage between non-transfected, transfected, serum-fed and serum-starved cells. However, the rates of GFP-expressing blastocysts were higher in the FuGENE 6 group (71.4%) among transfection reagents and in the fetal fibroblasts group (70.4%) for donor cells. These results indicate that fetal fibroblasts transfected with FuGENE 6 can be used as donor cells for porcine SCNT and that GFP gene can be safely used as a marker of foreign genes in porcine transgenesis.

• Mycobiology
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• v.40 no.1
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• pp.59-65
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• 2012

Antagonistic microorganisms against $Rhizoctonia$ $solani$ were isolated and their antifungal activities were investigated. Two hundred sixteen bacterial isolates were isolated from various soil samples and 19 isolates were found to antagonize the selected plant pathogenic fungi with varying degrees. Among them, isolate C9 was selected as an antagonistic microorganism with potential for use in further studies. Treatment with the selected isolate C9 resulted in significantly reduced incidence of stem-segment colonization by $R.$ $solani$ AG2-2(IV) in Zoysia grass and enhanced growth of grass. Through its biochemical, physiological, and 16S rDNA characteristics, the selected bacterium was identified as $Bacillus$ $subtilis$ subsp. $subtilis$. Mannitol (1%) and soytone (1%) were found to be the best carbon and nitrogen sources, respectively, for use in antibiotic production. An antibiotic compound, designated as DG4, was separated and purified from ethyl acetate extract of the culture broth of isolate C9. On the basis of spectral data, including proton nuclear magneric resonance ($^1H$ NMR), carbon nuclear magneric resonance ($^{13}C$ NMR), and mass analyses, its chemical structure was established as a stereoisomer of acetylbutanediol. Application of the ethyl acetate extract of isolate C9 to several plant pathogens resulted in dose-dependent inhibition. Treatment with the purified compound (an isomer of acetylbuanediol) resulted in significantly inhibited growth of tested pathogens. The cell free culture supernatant of isolate C9 showed a chitinase effect on chitin medium. Results from the present study demonstrated the significant potential of the purified compound from isolate C9 for use as a biocontrol agent as well as a plant growth promoter with the ability to trigger induced systemic resistance of plants.

• Journal of Embryo Transfer
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• v.8 no.2
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• pp.151-157
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• 1993

The present study was undertaken to determine the optimal condition for parthenogenetic activation of rabbit oocytes by electric stimulation in vitro in an attempt to develop nuclear transplantation techniques for cloning mammalian embryos and animals. Freshly ovulated oocytes were collected from superovulated rabbits from 13 to 26 hrs. after hCG injection. The cumulus-free oocytes were activated parthenogetically by repeated stimuli of square direct electric pulses in O.3M mannitol solution. After applying electric stimulations of different voltages, pulse durations and pulse times, all of the oocytes were cultured in TCM-199 with 10% FCS for 96 hours in a 5% $CO_2$ incubator, and their developmental potential in vitro was examined. The higher activation rate (68.9%) was achieved at the voltage of 2.0kv/cm, the pulse duration of 60 $\mu$sec and three pulse times and the activation rate of 100% was achieved at the pulse duration of 100 and 200 $\mu$sec, the voltage of 1.5kv/cm and three pulse times. Although the higher rates of activation of oocytes were achieved at 100 and 200 $\mu$sec, none of them developed to blastocyst in vitro. The oocytes collected 18~20 hours post hCG injection showed the highest rate of activation and development to blastocyst in vitro than the oocytes collected 13~15 or 25~26 hours post hCG injection. Therefore, it can be suggested that the application of electric stimulation of 2.0kv/cm, 60 $\mu$sec and three pulse times to the oocytes collected at 18~20 hours post hCG injection would be more beneficial for the parthenogenetic activation of oocytes in rabbits.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.1
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• pp.143-151
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• 2001

A marine bacterium producing carotenoid was isolated from the Yosu coastal area of South Korea, which was recorded as MCK-1. It was identified as Erythrobacter sp. Optimium conditions of marine carotenoid fermentation from Erythrobacter sp. were pH 6.0, a temperature of $25^{\circ}C$, 16 mM mannitol as a carbon source, 0.5% tryptone as a nitrogen source, 0.1 mM $Fe^{+2}$ ion as a mineral source and 1$\mu$M of cyanocobalamine as a growth factor in a jar-fermentor. Erythrobacter sp. was produced 351.27 mg/100mL of the marine carotenoid in these optimum conditions. This marine carotenoid was composed of 4 different conpounds, like as notoxanthin (61.4%), can thaxanthin (24.6%), fucoxanthin (8.2%), and zeaxanthin (5.8%). Physiological properties including antibacterial activity, cytotoxic effect, antioxidative effect and free radical scavenging activity were characterized with crude carotenoid. Carotenoid exhibited no antibacterial activity against E. coli and lactobacillus bulgaricus, but showed cytotoxic effect against cancer cells such as HepG2 (Hepatocellular carcinoma, human, ATCC HB-8065) and HeLa (Cervical carcinoma, human, ATCC CCL-2) cells. The impediment ratios for HepG2 and HeLa cell were 37.14% and 33.78%, respectively. This carotenoid expressed a strong antioxidative effect (77%) against CCL-13 5 $\mu\textrm{g}$/mL and 50 $\mu\textrm{g}$/mL crude carotenoid, respectively.

• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.127-131
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• 1997

Alkaline phosphatase (APase) was found to be inducible in Anabaena sp. strain CA Growth was less than control in presence of most amino acids except glycine and serine, but most amino acids enhanced APase activity. Highest APase activity was recorded in tyrosine supplemented culture followed by hydroxyproline, cystein, valine and glutamic acid. Threonine supplemented material showed lowest APase level (1.8 nmol/mg protein/min). Lactose, glucose, sodium pyruvate and succinate stimulated growth but not APase activity. APase activity was high in the presence of sucrose, mellibiose, mannitol, arabinose, maltose and sorbose, even though the growth in these supplements was less than in control. Organic phosphate sources supported good growth of the organism. Best growth occurred in presence of inorganic phosphate, adenosine diphosphate, fructose 1,6-diphosphate or ribulose 1,5-diphosphate, followed by other phosphorus sources tested. APase activity in presence of any of the organic phosphate sources was 3 to 5 fold low as compared to phosphate limited culture. Also, there was no APase activity in cultures grown on inorganic phosphate. These data indicate that most amino acids and a few carbohydrates (sucrose, mellibiose, arabinose and sorbose) are suitable for APase production. Lactose, glucose, pyruvate or succinate may be used as a carbon source during photoheterotrophic growth of the cyanobacterium. Glycine and serine are preferred nitrogen sources for its growth. Phosphate repressible APase activity has been found in Anabaena sp. strain CA.