• The Plant Pathology Journal
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• v.27 no.1
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• pp.59-67
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• 2011

A bacterial strain CH-67 which exhibits antagonism towards several plant pathogenic fungi such as Botrytis cinerea, Fulvia fulva, Rhizoctonia solani, Sclerotinia sclerotiorum, Colletotrichum sp. and Phytophthora sp. was isolated from forest soil by a chitin-baiting method. This strain was identified as Burkholderia cepacia complex (Bcc) and belonging to genomovar IX (Burkholderia pyrrocinia) by colony morphology, biochemical traits and molecular method like 16S rRNA and recA gene analysis. This strain was used to develop a bio-fungicide for the control of tomato leaf mold caused by Fulvia fulva. Various formulations of B. pyrrocinia CH-67 were prepared using fermentation cultures of the bacterium in rice oil medium. The result of pot experiments led to selection of the wettable powder formulation CH67-C containing modified starch as the best formulation for the control of tomato leaf mold. CH67-C, at 100-fold dilution, showed a control value of 85% against tomato leaf mold. Its disease control efficacy was not significantly different from that of the chemical fungicide triflumidazole. B. pyrrocinia CH-67 was also effective in controlling damping-off caused by Rhizoctonia solani PY-1 in crisphead lettuce and tomato plants. CH67-C formulation was recognized as a cell-free formulation since B. pyrrocinia CH-67 was all lethal during formulation process. This study provides an effective biocontrol formulation of biofungicide using B. pyrrocinia CH-67 to control tomato leaf mold and damping-off crisphead lettuce and tomato.

• Biomolecules & Therapeutics
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• v.2 no.4
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• pp.376-382
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• 1994

Repeated injections of low-doses of mercuric chloride in rats or mice induce polyclonal activation which includes the induction of anti-glomerular basement membrane (GBM) antibodies and circulating immune complex and it results in nephritis. Because this disease is autoimmune mediated disease resulted from immune dysfunction, immunomodulators are used to control the symptoms or to cure the disease. Irpex lacteus Fr. is a kind of new medicinal fungus. The polysaccharide fraction extracted from submerged fermentation of Irpex lacteus Fr. decreased the serum agglutinin, serolysin and IgM plaque forming cells in normal mice. The hitherto obtained clinical results suggested that it significantly improved the oligourea, edema, and hypertension in patients who have nephritis. To elucidate the action-mechanisms of Irpex lacteus Fr., we established the experimental model of HgCl$_2$induced polyclonal activation by intraperitoneal administrations of HgCl$_2$to mice. To assess the immunomodulating effect of Irpex lacteus fraction, we Investigated its effects on the mitogen induced proliferation and IgM PFC counts of splenic lymphocytes in mice during the treatment of HgCl$_2$. The Irpex lacteus polysaccharide reduced the abnormally increased mitogen induced Iymphocyte proliferation and IgM PFCs to almost normal levels. And the Irpex lacteus polysaccharides prevented the increasement of serum immunoglobulin level induced by HgCl$_2$. These data suggested that the Irpex lacteus polysaccharides might have the immunomodulating activity to prevent and /or improve the HgCl$_2$ induced autoimmune disease.

• Journal of Animal Science and Technology
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• v.65 no.2
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• pp.387-400
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• 2023

Ruminal protozoa, especially entodiniomorphs, engulf other members of the rumen microbiome in large numbers; and they release oligopeptides and amino acids, which can be fermented to ammonia and volatile fatty acids (VFAs) by amino acid-fermenting bacteria (AAFB). Studies using defaunated (protozoa-free) sheep have demonstrated that ruminal protozoa considerably increase intraruminal nitrogen recycling but decrease nitrogen utilization efficiency in ruminants. However, direct interactions between ruminal protozoa and AAFB have not been demonstrated because of their inability to establish axenic cultures of any ruminal protozoan. Thus, this study was performed to evaluate the interaction between Entodinium caudatum, which is the most predominant rumen ciliate species, and an AAFB consortium in terms of feed degradation and ammonia production along with the microbial population shift of select bacterial species (Prevotella ruminicola, Clostridium aminophilum, and Peptostreptococcus anaerobius). From an Ent. caudatum culture that had been maintained by daily feeding and transfers every 3 or 4 days, the bacteria and methanogens loosely associated with Ent. caudatum cells were removed by filtration and washing. An AAFB consortium was established by repeated transfers and enrichment with casamino acids as the sole substrate. The cultures of Ent. caudatum alone (Ec) and AAFB alone (AAFB) and the co-culture of Ent. caudatum and AAFB (Ec + AAFB) were set up in three replicates and incubated at 39℃ for 72 h. The digestibility of dry matter (DM) and fiber (NDF), VFA profiles, ammonia concentrations, pH, and microscopic counts of Ent. caudatum were compared among the three cultures. The co-culture of AAFB and Ent. caudatum enhanced DM degradation, VFA production, and Ent. caudatum cell counts; conversely, it decreased acetate: propionate ratio although the total bacterial abundance was similar between Ec and the Ec + AAFB co-culture after 24 h incubation. The ammonia production and relative abundance of C. aminophilum and P. anaerobius did not differ between AAFB alone and the Ec + AAFB co-culture. Our results indicate that Ent. caudatum and AAFB could have a mutualistic interaction that benefited each other, but their interactions were complex and might not increase ammoniagenesis. Further research should examine how such interactions affect the population dynamics of AAFB.

• Animal Bioscience
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• v.37 no.2_spc
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• pp.370-384
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• 2024

Rumen microbiota play a central role in the digestive process of ruminants. Their remarkable ability to break down complex plant fibers and proteins, converting them into essential organic compounds that provide animals with energy and nutrition. Research on rumen microbiota not only contributes to improving animal production performance and enhancing feed utilization efficiency but also holds the potential to reduce methane emissions and environmental impact. Nevertheless, studies on rumen microbiota face numerous challenges, including complexity, difficulties in cultivation, and obstacles in functional analysis. This review provides an overview of microbial species involved in the degradation of macromolecules, the fermentation processes, and methane production in the rumen, all based on cultivation methods. Additionally, the review introduces the applications, advantages, and limitations of emerging omics technologies such as metagenomics, meta-transcriptomics, metaproteomics, and metabolomics, in investigating the functionality of rumen microbiota. Finally, the article offers a forward-looking perspective on the new horizons and technologies in the field of rumen microbiota functional research. These emerging technologies, with continuous refinement and mutual complementation, have deepened our understanding of rumen microbiota functionality, thereby enabling effective manipulation of the rumen microbial community.

• Applied Biological Chemistry
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• v.39 no.3
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• pp.195-199
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• 1996

We have cloned several E. coli sfs genes which stimulate mal gene expression with $crp^{{\ast}1}$). One the genes (pPVC2) was sequenced and potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. In order to investigate the regulation of the sfs1 gene by the cAMP-CRP complex, we have constructed the sfs-lacZ fusion gene in this research. The overall transcriptional stimulations of sfs1 gene in the presence cAMP were confirmed by ${\beta}-galactosidase$ activity and Western blot analysis of sfs1-lacZ fusion gene. Transcriptional regulation by cAMP-CRP was also confirmed by Northern blot analysis. End-labelled DNA of the DNA fragment in sfs1 regulation region were used for gel retardation assay to examine the CRP-DNA complex in the presence of cAMP. Results here indicate that CRP binding site in the regulatory region of sfs1 gene is positive regulator for the expression of sfs1 gene.

• Journal of Life Science
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• v.34 no.7
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• pp.465-475
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• 2024

Lactiplantibacillus plantarum K9 is a probiotic strain that can be utilized from various bioactive substances isolated from Protaetia brevitarsis seulensis larvae. In this study, a genetic analysis of L. plantarum K9 revealed the existence of a bacterial chromosome and three plasmids. The glycolysis pathway and pentose phosphate pathway were examined for their normal functioning via an analysis of the core metabolic pathways of L. plantarum K9. Since the key enzymes, fluctose-1,6-bisphospatase (EC: 3.1.3.11) and 6-phosphogluconate dehydratase (EC: 4.2.1.12)/2-keto-deoxy-6-phosphogluconate (KDPG) aldolase (EC: 4.2.1.55), of gluconeogenesis and the ED pathway were not identified from the L. plantarum K9 genome, we suggest that gluconeogenesis and the ED pathway are not performed in L. plantarum K9. Additionally, while some enzymes, related to fumarate and malate biosyntheses, involved in the TCA cycle were identified from L. plantarum K9, the enzymes associated with the remaining TCA cycle were absent, indicating that the TCA cycle cannot proceed. Meanwhile, based on our findings, we propose that the oxidative electron transport system performs class IIB-type (bd-type) electron transfer. In summary, we assert that L. plantarum K9 performs homolactic fermentation, executes gluconeogenesis and the pentose phosphate pathway, and carries out energy metabolism through the class IIB-type oxidative electron transport system. Therefore, we suggest that L. plantarum K9 has relatively high lactic acid production, and that it has excellent antibacterial activity, as a result, compared to other lactic acid bacterial strains. Moreover, we speculate that L. plantarum K9 has an oxidative electron transport capability, indicating that it is highly resistant to oxygen and suggesting that it has fine cultivation characteristics, which collectively make it highly suitable for use as a probiotic.

• KSBB Journal
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• v.15 no.5
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• pp.489-496
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• 2000

Several culture systems including batch, two-stage CSTR, semi-fed batch, and two-stage cyclic fed-batch were investigated for the efficient production of the Fab fraction of PDC-E2 specific human monoclonal antibody using high cell density recombinant E. coli. A two-phase batch system and a two-stage continuous system were examined to overcome plasmid instability problems, by separating the growth and the production stages. The cell density and productivity of the two-stage continuous culture was better than that of the two-phase batch fermentation. In the two-stage continuous culture system with DO-stat, the cell growth and the productivity were superior to those of the system without the DO control. Also, almost total plasmid stability was maintained in the two-stage continuous culture system. Modified M9 medium was selected as an optimum feeding medium for the fed-batch process, and the optimum C/N ratio determined to be 2:3. The optimum feeding rate was $0.6g/\ell/hr$ for a constant feeding strategy in semi-fed batch system. When the feeding medium was fed by pulsing, it was observed that more frequent pulsing resulted in improved cell growth. The linear feeding method was the most efficient of the various feeding methods tested. Finally, high cell density culture using a two-stage cyclic fed batch system with pH-stat was tried because the linear feeding method showed limitations in terms of obtaining high cell densities, and a cell density of $54 g/\ell$ was achieved. It was concluded that the two-stage cyclic fed batch system was the most efficient system for high cell density culture of the systems tested. However, productivity improvements were lower than expected due to the extremely high accumulations of acetate, although the low levels of residual glucose were maintained.

• Korean Journal of Environmental Agriculture
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• v.34 no.2
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• pp.91-97
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• 2015

BACKGROUND: Earthworms are essential detritus feeders that play a vital role in the process of decomposition of organic matter and soil metabolism. The complex process of partial breakdown of organic matter and mixing with mucous and gut microbial flora in the form of earthworm cast results in the reduction of the toxicity. This study focused on the change of cast amount and pollutant contents before and after the eating of the organic waste and upland soil with the two species of earthworm. METHODS AND RESULTS: The two species of earthworms were compared to the cast production. In the upland soil material, the daily amount of worm's cast was 1.42 g in E. andrei and 0.40 g in A. agrestis. In the organic waste material, the cast of E. andrei was 0.78~0.83 g and the cast of A. agrestis. have not been collected because all earthworms died after the treatment. The heavy metals treated in the upland soil were evaluated the impact of the worm excretion. With the E. andrei, the cast production was decreased 0.1~0.8 times in zinc, 0.2~0.5 times in copper, and 0.1~0.7 times in cadmium compared to the control treatment according to the levels of concentration. With A. agrestis, the cast amount was decreased 0.3~1.1 times in zinc, 0.2~0.3 times in copper, and 0.1~2.1 times in cadmium, respectively. The changes of pollutant contents before and after the eating of the organic wastes with E. andrei were studied. In the treatment of the Alcohol Fermentation Processing Sludge and the Fruit Juice Processing Sludge, heavy metal content of the cast was increased 0.7~53.3% compared to the sludge materials. PAHs contents were decreased 50.1% in the cast of the Alcohol Fermentation Processing Sludge and 36.6% in the cast of the Fruit Juice Processing Sludge, respectively. CONCLUSION: In conclusion, although the A. agrestis was bigger than E. andrei in size and weight, the cast amount of A. agrestis was small. The two species of earthworm was less excretion with high concentration of heavy metals. While the heavy metals such as zinc, copper, and cadmium were considerably accumulated in the cast, the total compounds, PAHs were fairly decomposed. There results would provide us for restoring contaminated soil and cleaning organic wastes.

• Microbiology and Biotechnology Letters
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• v.41 no.3
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• pp.327-334
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• 2013

For the development of hardy kiwi wine, we arranged for the post-maturity of hardy kiwi fruit, treated them with calcium carbonate and a pectinase enzyme complex, investigated the resulting physicochemical properties and conducted a sensory evaluation. The period determined for creating post-maturity in the hardy kiwi fruit was determined as 5 days storage at room temperature following maturity. During this time the yield of fruit juice was increased from 22.1% to 53.5% using 0.1% (v/v) cytolase PCL5 for 2 h at room temperature. 0.1% (w/v) calcium carbonate was also added during the process of aging, for the reduction of the sour taste. The fermentation trial of the hardy kiwi wine was prepared using water (25% or 50%), sugar ($24^{\circ}brix$), 0.1% (w/v) $CaCO_3$, 0.1% (v/v) cytolase PCL5, $K_2S_2O_5$ (200 ppm), and yeast ($1.5{\times}10^7$ cell/ml). Fermentation then occurred for 2 weeks at $20^{\circ}C$. The pH value, total acidity, alcohol, and reducing sugar content of the resulting hardy kiwi wines of 25% (v/w) and 50% (v/w) water, were in a range of pH 3.4-3.7, 1.12-1.21%, 14.3-14.4%, and 15-16 g/l, respectively. Citric acid and fructose constituted the major organic acids and the free sugar of the 25% and 50% hardy kiwi wine, respectively. Volatile flavor components, including 10 kinds of esters, 8 kinds of alcohols, 5 kinds of acids, 3 kinds of others and aldehydes, were determined by GC analysis. The results of sensory evaluation demonstrated that 50% hardy kiwi wine is more palatable than 25% hardy kiwi wine.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2005.06a
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• pp.154-164
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• 2005

Saccharomyces cerevisiae KNU5377 is a thermotolerant strain, which can ferment ethanol from wasted papers and starch at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. This strain showed alcohol fermentation ability to convert wasted papers 200 g (w/v) to ethanol 8.4% (v/v) at 40$^{\circ}C$, meaning that 8.4% ethanol is acceptable enough to ferment in the industrial economy. As well, all kinds of starch that are using in the industry were converted into ethanol at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. Hyperthermic cell killing kinetics and differential scanning calorimetry (DSC) revealed that exponentially growing cells of this yeast strain KNU5377 were more thermotolerant than those of S. cerevisiae ATCC24858 used as a control. This intrinsic thermotolernace did not result from the stability of entire cellular components but possibly from that of a particular target. Heat shock induced similar results in whole cell DSC profiles of both strains and the accumulation of trehalose in the cells of both strains, but the trehalose contents in the strain KNU5377 were 2.6 fold higher than that in the control strain. On the contrary to the trehalose level, the neutral trehalase activity in the KNU5377 cells was not changed after the heat shock. This result made a conclusion that though the trehalose may stabilize cellular components, the surplus of trehalose in KNU5377 strain was not essential for stabilization of whole cellular components. A constitutively thermotolerant yeast, S. cerevisiae KNU5377, was compared with a relatively thermosensitive control, S. cerevisiae ATCC24858, by assaying the fluidity and proton ATPase on the plasma membrane. Anisotropic values (r) of both strains were slightly increased by elevating the incubation temperatures from 25$^{\circ}C$ to 37$^{\circ}C$ when they were aerobically cultured for 12 hours in the YPD media, implying the membrane fluidity was decreased. While the temperature was elevated up to 40$^{\circ}C$, the fluidity was not changed in the KNU5377 cell, but rather increased in the control. This result implies that the plasma membrane of the KNU5377 cell can be characterized into the more stabilized state than control. Besides, heat shock decreased the fluidity in the control strain, but not in the KNU5377 strain. This means also there's a stabilization of the plasma membrane in the KNU5377 cell. Furthermore, the proton ATPase assay indicated the KNU5377 cell kept a relatively more stabilized glucose metabolism at high temperature than the control cell. Therefore, the results were concluded that the stabilization of plasma membrane and growth at high temperature for the KNU5377 cell. Genome wide transcription analysis showed that the heat shock responses were very complex and combinatory in the KNU5377 cell. Induced by the heat shock, a number of genes were related with the ubiquitin mediated proteolysis, metallothionein (prevent ROS production from copper), hsp27 (88-fold induced remarkably, preventing the protein aggregation and denaturation), oxidative stress response (to remove the hydrogen peroxide), and etc.