• The Plant Pathology Journal
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• 제35권2호
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• pp.91-99
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• 2019

Mitogen-activated protein kinase (MAPK) cascades in fungi are ubiquitously conserved signaling pathways that regulate stress responses, vegetative growth, pathogenicity, and many other developmental processes. Previously, we reported that the AbSte7 gene, which encodes a mitogen-activated protein kinase kinase (MAPKK) in Alternaria brassicicola, plays a central role in pathogenicity against host cabbage plants. In this research, we further characterized the role of AbSte7 in the pathogenicity of this fungus using ${\Delta}AbSte7$ mutants. Disruption of the AbSte7 gene of A. brassicicola reduced accumulation of metabolites toxic to the host plant in liquid culture media. The ${\Delta}AbSte7$ mutants could not efficiently detoxify cruciferous phytoalexin brassinin, possibly due to reduced expression of the brassinin hydrolase gene involved in detoxifying brassinin. Disruption of the AbSte7 gene also severely impaired fungal detoxification of reactive oxygen species. AbSte7 gene disruption reduced the enzymatic activity of cell walldegrading enzymes, including cellulase, ${\beta}$-glucosidase, pectin methylesterase, polymethyl-galacturonase, and polygalacturonic acid transeliminase, during host plant infection. Altogether, the data strongly suggest the MAPKK gene AbSte7 plays a pivotal role in A. brassicicola during host infection by regulating multiple steps, and thus increasing pathogenicity and inhibiting host defenses.

• The Plant Pathology Journal
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• 제37권4호
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• pp.356-364
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• 2021

This study was to investigate defense mechanisms on cassava induced by salicylic acid formulation (SA) against anthracnose disease. Our results indicated that the SA could reduce anthracnose severity in cassava plants up to 33.3% under the greenhouse condition. The 𝛽-1,3-glucanase and chitinase enzyme activities were significantly increased at 24 hours after inoculation (HAI) and decrease at 48 HAI after Colletotrichum gloeosporioides challenge inoculation, respectively, for cassava treated with SA formulation. Synchrotron radiation-based Fourier-transform infrared microspectroscopy spectra revealed changes of the C=H stretching vibration (3,000-2,800 cm^-1), pectin (1,740-1,700 cm^-1), amide I protein (1,700-1,600 cm^-1), amide II protein (1,600-1,500 cm^-1), lignin (1,515 cm^-1) as well as mainly C-O-C of polysaccharides (1,300-1,100 cm^-1) in the leaf epidermal and mesophyll tissues treated with SA formulations, compared to those treated with fungicide carbendazim and distilled water after the challenged inoculation with C. gloeosporioides. The results indicate that biochemical changes in cassava leaf treated with SA played an important role in the enhancement of structural and chemical defense mechanisms leading to reduced anthracnose severity.

• 대한약침학회지
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• 제25권4호
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• pp.317-325
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• 2022

Polymers are the major constructive material of pharmaceutical formulations that play a prime role in designing effective drug-delivery systems and releasing drugs at their sites of application. Polymers are composed of multiple repeating units of high molecular mass components with attendant properties. Most synthetic polymers are non-biocompatible, expensive, and extremely inclined to deliver adverse impacts. Meanwhile, edible polymers obtained from natural sources have gained remarkable recognition for their promising use in modern medicine. Moreover, polymers derived from natural sources are generally preferred due to certain of their unique features such as abundant availability, biocompatibility, nontoxicity, economical, safe, and effective functions that fit the purpose. Polysaccharides including starch, cellulose, hemicellulose, pectin, and mucilage are identified as a major class of naturally obtained molecules that have a substantial role as functional polymers. This review summarizes the potential role of polysaccharides derived from vegetable sources such as adhesives, anticaking agents, binders, disintegrants, emulsifiers, film-framing agents, and thickeners. This is simply an opportunity to abandon synthetic excipients that hurt our bodies and think back to nature from where we originate.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.204-204
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• 2022

Transformant construction using protoplasts requires less sample preparation time than particle bombardment and Agrobacterium-mediated transfection. There are two protoplast transfection methods: the PEG-mediated transfection method and the Lipofectamine transfection method. When Lipofectamine is mixed with DNA, Lipofectamine surrounds DNA like a cell membrane because of the positive charge of Lipofectamine. The Lipofectamine-DNA complex makes DNA insertion into cells easier. Fectin has similar functions to lipofectamine and is less expensive than lipofectamine. The 3D-fectin technology has been highlighted in animal cell transfection. Therefore, we performed PEG-mediated transfection, Lipofectamine transfection, and 3D-pectin transfection with a GFP construct. Protoplasts were isolated using the first leaf of "Bobwhite" after 4 hours of incubation in an isolation Buffer (cellulase + macerozyme). Protoplasts transformed by each method were cultured for 48 hours, and then GFP fluorescence expression was confirmed under confocal microscopy. GFP signals were detected in PEG-mediated transfection and Lipofectamine transfection. And the GFP signals were also detected in protoplasts to which 3D-fectin technology was applied, suggesting that 3D-fectin technology can be used for plant protoplast transfection.

• Journal of Applied Biological Chemistry
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• 제55권3호
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• pp.141-148
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• 2012

최근 주류 섭취 시 숙취와 관련이 있는 성분인 아세트알데히드, 메탄올 및 퓨젤유를 포함한 휘발성 유해성분에 대한 관심이 증가하고 있다. 또한, 이들 성분들은 과음 시 간질환 및 암을 유발할 수도 있는 것으로 알려져 있다. 아세트알데히드는 주류에서 발견되는 휘발성 성분이며 많은 식품에서 향미소재로 사용되고 있으나, 인체에 발암가능 물질로 분류되기도 한다. 특히, 알코올과 같이 섭취 시에는 1급 발암물질로 분류되고 있다. 메탄올은 알코올 발효 중 펙틴 분해 효소에 의해 팩틴의 demethoxylation 기작으로 생성된다. 이에 비해 퓨젤유는 알코올 발효의 부산물로 생성되며, 주류의 주요한 향미성분으로 알려져 있다. 본 연구에서는 주류에 포함되어 있는 휘발성 유해성분들이 건강에 미치는 영향, 이들의 최대허용함량 및 다양한 주류에서의 실제 함량, 그리고, 이들의 분석방법에 대해 고찰하였다.

• 동아시아식생활학회지
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• 제13권6호
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• pp.601-607
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• 2003

This study was attempted to investigate the physicochemical changes during soybean-leaf water kimchis fermentation by adding wheat flour in cooking water. Soybean-leaf water kimchis with five different levels of wheat flour in cooking water(0%;A, 5%;B 10%;c, 15%;D, 20%;E) were tested for rhological parameters, fine structural changes, sensory evaluations and chemical analysis. The composition of the soybean-leaf was moisture (80.9%), protein(6.8%), fat(0.6%), ash(2.1%) and alkalinity (+14.9). The amount of reducing sugars of sample A and other samples were 0.75% and 1.08∼1.4% in the initial fermentation stage, but decreased to 0.3 and 0.43∼0.50% in the later fermentation stage, respectively. The pH of sample A decreased from 5.17 to 4.72 during the initial fermentation. On sample B, C, D and E, pH's decreased rapidly during the initial fermentation, but they did not change much in the later fermentation stage. The pectin contents of all samples decreased during 2nd and 4th day of fermentation, then the change was slow. The amount of hemicellulose. cellulose and lignin in terms of the DNF and ADF were varied from 4 to 33%, but the contents of them did not greatly changed during the fermentation. The sensory evaluation showed that both B and C samples had the good score in sweety taste, roasted nutty taste, and the ease of swallowing measured as chewiness. From these results, the optimum soybean-leaf water kimchis can be prepared when 200g soybean-leaf, 1000$m\ell$ water, 15g garlic, 3% red pepper powder and 5 ∼0% wheat flour were fermented at 20$^{\circ}C$ for 2 days.

• 펄프종이기술
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• 제45권3호
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• pp.59-68
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• 2013

The cotton contains various impurities such as protein, wax, pectins, coloring matter, even though cotton has been a major source of pure cellulose. The purification processes have been commonly applied to obtain the pure cellulose. However the excessive purification treatments could lead to the damage in the cellulose structure which could result in the degradation of cellulose and the limited application of cotton cellulose. In this study, the changes in cellulose structural properties such as crystallinity and DP(degree of polymerization) by the various conditions of the purification processes were investigated. The less toxic agents such as hydrogen peroxide and sodium silicate were applied for the purification treatment in this study. The increase in the process times, the temperature and the applied amount of chemical agents resulted in the more purified cellulose. The DP of cotton cellulose was increased at the first weak conditions by the reduction of small molecules such as pectin, wax, and so on. Especially the 2 % addition amount of $H_2O_2$ with $Na_2SiO_3$ resulted in the higher value in the DP and the brightness compared to the 1.5 % addition amount of $H_2O_2$. However, the 4 % addition amount of $H_2O_2$ with $Na_2SiO_3$ showed the decreased value because of excessive treatment. In case of the changes in the crystallinity (Gjk), the highest value of the crystallinity was obtained by the 2% addition amount of $H_2O_2$ on the cotton cellulose, which showed similar with the change in the DP.

• 한국축산식품학회지
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• 제37권5호
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• pp.787-791
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• 2017

Panax ginseng marc is produced from fresh ginseng roots during processing and is generally treated as industrial waste. The primary aim of this study was to improve its utilization in the dairy industry as a potential high-value resource. Yogurt was prepared from 11% skim milk powder, 0.1% pectin, 10% sucrose, and ginseng marc ethanol extract (GME, 0.5% and 1.0%) in milk, and was inoculated with a 0.02% yogurt culture (Lactobacillus acidophilus, Bifido-bacterium longum, and Streptococcus thermophilus). After fermentation at $40^{\circ}C$ for 6-8 h, the physicochemical properties of samples were analyzed by the AOAC, Kjeldahl, and Soxhlet methods. Sensory evaluation was performed based on consumer acceptability scores with a 7-point scale, and antimicrobial effects were measured by the agar plate method. The moisture, crude protein, crude fat, and ash contents of yogurt supplemented with 1% GME were $85.06{\pm}0.06%$, $4.41{\pm}0.01%$, $4.30{\pm}0.05%$, and $0.81{\pm}0.03%$, respectively, with no significant changes noted from those of yogurt without GME (control), except for an increase in the crude fat content. The sensory scores of color, flavor, texture, overall taste, and overall acceptance of yogurt supplemented with below 1% GME did not differ significantly (p<0.05) to those of the control yogurt. In addition, the growths of Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes, Escherichia coli, and Enterobacter sakazakii were inhibited during fermentation and storage. These results suggest that GME could be used in dairy products as a supplement and in the food industry as an antimicrobial material.

• Journal of Microbiology and Biotechnology
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• 제27권4호
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• pp.791-807
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• 2017

The type II secretion system (T2SS), which transports selected periplasmic proteins across the outer membrane, has rarely been studied in nonpathogens or in organisms classified as Betaproteobacteria. Therefore, we studied Cupriavidus metallidurans (Cme), a facultative chemilithoautotroph. Gel analysis of extracellular proteins revealed no remarkable differences between the wild type and the T2SS mutants. However, enzyme assays revealed that native extracellular alkaline phosphatase is a T2SS substrate, because activity was 10-fold greater for the wild type than a T2SS mutant. In Cme engineered to produce three Ralstonia solanacearum (Rso) exoenzymes, at least 95% of their total activities were extracellular, but unexpectedly high percentages of these exoenzymes remained extracellular in T2SS mutants cultured in rich broth. These conditions appear to permit an alternative secretion process, because neither cell lysis nor periplasmic leakage was observed when Cme produced a Pectobacterium carotovorum exoenzyme, and wild-type Cme cultured in minimal medium secreted 98% of Rso polygalacturonase, but 92% of this exoenzyme remained intracellular in T2SS mutants. We concluded that Cme has a functional T2SS despite lacking any abundant native T2SS substrates. The efficient secretion of three foreign exoenzymes by Cme is remarkable, but so too is the indication of an alternative secretion process in rich culture conditions. When not transiting the T2SS, we suggest that Rso exoenzymes are probably selectively packaged into outer membrane vesicles. Phylogenetic analysis of T2SS proteins supports the existence of at least three T2SS subfamilies, and we propose that Cme, as a representative of the Betaproteobacteria, could become a new useful model system for studying T2SS substrate specificity.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.84.2-85
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• 2003

Magnaporthee grisea causes the devastating blast disease of rice. Entensive research has been conducted on infection mechanisms, particularly on appressorium formation and penetration, of this fungus during the last decade. However, the role(s) of cell-wall-degrading enzymes (CWDEs) on pathogenesis is not clearly demonstrated at molecular level. Many CWDES in plant pathogenic fungi including M. grisea are redundant; that is, there are multiple genes encoding enzymes with a similar or overlapping spectrum of activities. It is laborious to isolate all of the genes encoding related enzymes and to construct mutants lacking all 9f them. Thus, we considered alternative strategies to address the role of CWDEs in pathogenesis. Since expression of CWDE genes Is repressed by a simple sugar, as the first step, we cloned a Snfl (sucrose non-fermenting) gene (MgSnf1) from M. grisea. The predicted amino acid sequence showed a high identity with other Snf1 genes from various fungi. To elucidate molecular function of MgSnf1, a transformant lacking MgSnf1 was created by targeted gene replacement. En glucose, sucrose, and xylan the MgSnf1 mutant grew normally but in pectin and complex media, it grew slower than wild type. Expression of various CWDEs in MgSnf1 mutant was investigated and found that expression of some CWDEs is repressed. However, no significant difference was observed in conidial germination, appressorium formation, and pathogenicity in MgSnf1 mutant. However, MgSnf1 functionally complemented a yeast MgSnf1 mutant. These results suggest that MgSnf1 is involved in regulation of CWDEs and MgSnf1 is dispensable in pathogenicity of M. grisea.