• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.311-317
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• 2006

Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

• Food Science and Preservation
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• v.8 no.1
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• pp.66-73
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• 2001

This study was carried out to know whether $\beta$-galactosidase is directly important or not on fruit softening during the development of peach fruits compared to those in the stage stage. It was investigated that the flesh firmness, cell wall components, and the glycosidase activities of the peach fruits with a fast softening cultivar, 'Mibeakdo', a slow softening cultivar,'Yumyung'and a middle softening cultivar, 'Okubo$\beta$, at different developmental stages, on 13 May, 16 June, 16 July, and 5 August and on 28 August which harvested only 'Yumyung' fruits. In order to investigate the amounts of total sugar and non-cellulosic neutral sugar, the cell wall materials of each fruit were solubilized in distilled water, 0.05M CDTA, 0.05M Na$_2$CO$_3$, 4% KOH, and 24% KOH sequentially. During the fruit development, the fruit firmness of three cultivars decreased and the fruit firmness of 'Yumyung' was higher than that fo 'Mibeakdo' and 'Okubo' in the overall period. During the fruit development, the changes of total sugar amounts of each measured fractions were similar among peach cultivars. Arabinose and galactose were the predominant non-cellulosic neutral sugars in all the fractions including cell wall material of the three cultivars. There was an active relationship between the changes of flesh firmness in three cultivars and the mol % changes of rhamnose on 5 August which was the harvest date of 'Mibeakdo' and 'Okubo' fruits. The activity of soluble $\beta$-galactosidase was high at the early developmental stage and then dropped to a very low activity level in all cultivars. The activity of cell wall-bound $\beta$-galactosidase was high at the early developmental stage and then decreased continuously through the harvest date. In addition the changes of other glycosidase activities were similar among cultivars.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.377-384
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• 2009

Among 27 Bacillus sp. isolated from Meju, a traditional Korean soybean fermented food, a strain MJ-226 was selected due to its strong fibrinolytic activity, and it was identified to be Bacillus subtilis MJ-226 according to morphological and biochemical characterization and sugar utilization. The fibrinolytic enzyme of B. subtilis MJ-226 was maximally produced by cultivating in the Tryptic Soy Broth (TSB) for 24~26 h at $37^{\circ}C$, and the enzymes activity was promoted with adding glucose, fructose, peptone or yeast extract to TSB. The fibrinolytic enzyme was stable at the range of pH from 6.0 to 8.0, and between 35 and $40^{\circ}C$. Also, when the crude enzyme was exposed to various metal ions and chemical inhibitors for 12 h, the enzyme stability was maintained by $MnSO_4$, $CaCl_2$, KCl, and NaCl. However, the stability was destroyed by treatment with $CuSO_4$, $MgSO_4$, $ZnSO_4$, $FeSO_4$, and $BaCl_2$, and the enzyme was unstable in the presence of chemical inhibitors such as iodoacetic acid, leupeptin, phenylmethanesulphonyl fluoride (PMSF), sodium dodecyl sulfate (SDS), thiourea, trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA) and ethylenediaminetetraacetic acid (EDTA).

• Analytical Science and Technology
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• v.17 no.6
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• pp.454-463
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• 2004

The sorption behavior of some metal ions on XAD-16-CTA chelating resin was investigated by batch method. The sorption of chelating resin was highly selective for Hf(IV), Zr(IV) and Th(IV) at pH 3.0 ~ 6.0 and the maximum sorption capacity of Zr(IV) ion was 0.81 mmol/g. It was successfully applied to the separation of several rare metal ions from mixed metal solutions by using CDTA, EDTA, NTA and $NH_4F$ as masking agent. The elution order of metal ions obtained from breakthrough capacity and the overall capacity at pH 4.0 was Zr(IV)>Th(IV)>Hf(IV)>U(VI)>Cu(II)>In(III)>Pb(II). Desorption characteristics for metal ions was investigated with desorption agents such as HCl, $HNO_3$, $HClO_4$. 2 M HCl showed high desorption efficiency. Th(IV) ion can be successfully separated from mixed metal ions by using XAD-16-CTA cheating resin.

• Preventive Nutrition and Food Science
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• v.17 no.4
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• pp.293-298
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• 2012

In this study, we designed a method to manufacture elevated fermented green tea by using Monascus pilosus, which is known as a functional microbe, and observe its antioxidant abilities and quality characteristics. The water-soluble substance (WSS) content of the fermented tea by M. pilosus (FTM) was lower than that of the non-fermented tea (NFT), although the alcohol-insoluble substance (AIS) content of the FTM was higher than that of NTM. On the other hand, the fractionated distilled water-soluble polysaccharide (DWSP), CDTA-soluble polysaccharides (CDSP), sodium carbonate-soluble polysaccharide (SCSP) and KOH soluble hemicellulose (HC) obtained from the AIS of the FTM was markedly higher than that of NFT. In the antioxidant parameters, the electron donating ability of all fractions, except HC, extracted from FTM was higher than that of NFT, and iron chelating ability of all fractions, except CDSP, extracted from FTM was higher than that of NFT. Whereas the DWSP and SCSP obtained from the FTM were higher than that of NFT, the activity of the HC fraction from both NFT and the FTM could not be detected. In addition, the xanthin oxidase (XO) inhibitory activities of the DWSP, CDSP and the SCSP obtained from the NFT were significantly higher than that of FTM, the aldehyde oxidase (AO) inhibitory activities of the DWSP and SCSP extracted from the FTM were markedly higher than that of the NFT. Meanwhile, the acceptance of NFT and FTM had no significant difference, while the quality of aroma, taste and mouthfeel of the FTM was higher than that of NFT. These results suggest that the post-fermented tea by Monascus microorganisms may be responsible for functional components as well as contribute to the improvement of the tea quality.

• Journal of Periodontal and Implant Science
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• v.37 no.sup2
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• pp.339-351
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• 2007

Cytolethal distending toxin(CDT)은 세포 주기 중 G2에서 M 기로의 전환을 막아 세포의 증식을 억제할 수 있는 세균 단백 독소의 일종이다. 구강 미생물 중 유일하게 Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans)만이 이 CDT를 생성 할 수 있는 것으로 알려져 있다. A. actinomycetemcomitans는 localized aggressive periodontitis (LAP)의 원인균으로 여겨지며 비 운동성의 그람 음성 구간균이고 $37^{\circ}C$, 5% $CO_2$ 하에 성장이 왕성하다. A. actinomycetemcomitans의 CDT는 3개의 인접한 유전자인 cdtA, cdtB, cdtC에 의해 형성 되며 각각의 유전자에 대한 단백질의 기능은 아직 완전히 밝혀지지 않았다. 현재까지 연구에 의하면 cdtA는 CDT의 세포부착과 관련이 있는 것으로 여겨지며 이 유전자의 기능 이상 시 CDT의 독성 효과가 현저히 감소한다고 알려져 있다. 따라서 본 연구는 A. actinomycetemcomitans의 cdtA 유전자를 클로닝, 단백질 발현하여 향후 치주질환의 발병 과정에서 CdtA의 역할을 규명하고 질환의 예방 및 치료법에 도움을 주고자 하였다. A. actinomycetemcomitans Y4균주를 cdtA 유전자 클로닝을 위해 사용하였다. A. actinomycetemcomitans의 genomic DNA는 genomic DNA 추출 kit를 사용하여 분리하고 cdtA에 특이적인 primer를 이용하여 PCR을 통해 cdtA 유전자를 증폭하였다. 증폭된 cdtA 유전자를 T-vector에 클로닝 하였으며, 클로닝 된 cdtA 유전자는 단백질 발현을 위해 pRSET Avector에 서브클로닝 한 후 발현 균주인 BL21(DE3)를 이용하여 발현시켰다. 발현 후 Ni-NTA AP conjugate를 이용한 Western blot을 통해 pRSET-CDTA를 확인하였다.

• Journal of Bio-Environment Control
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• v.15 no.1
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• pp.91-99
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• 2006

This study was conducted to determine the influence of postharvest dipping treatment with aminoethoxyvinylglycine (AVG) on ethylene production and composition of non-cellulosic neutral sugars in cell walls of 'Tsugaru' apple fruits during storage. Fruits were harvested on August 20, soaked in AVG 50 and 75 $mg L^{-1}$ solution for 5 minutes, and stored in cold storage chamber at $0{\pm}1^{\circ}C$ for 60 days. Fruit quality factor, ethylene productions, and cell wall component changes were investigated at 20 days interval. As a result, the fruit firmness and acid content were much higher in AVG treated fruits than those of untreated one during 60 days of cold storage. Ethylene production of AVG treated fruits was reduced to the level of 1/10 compared with untreated one. As to the change of non-cellulosic neutral sugars in the cell walls of 'Tsu- garu' fruits, the major sugar was arabinose and galactose in water, CDTA and $Na_2CO_3$ soluble fractions. The content of arabinose and galactose in untreated fruits increased as the softening of fruits was in progress, but the fruits treated with AVG showed a little change during storage, so it is predicted that these two cell wall compositional sugars were not solubilized by the treatment of AVG. Accordingly, the marketability of 'Tsu- garu' fruits could remarkably increase when soaking the fruits in AVG solution after harvest.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 2003.10a
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• pp.134.1-134
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• 2003

The purpose of this study was to investigate the release of p-hydroxybenzoic acid and other compounds from cell wall materials(CWM) and their cellulose fraction from carrot with chemical and enzymatic hydrolysis. To investigate this effect on cell wall chemistry of carrot, alcohol insoluble residue(AIR) of CWM were prepared and were extracted sequentially with water, imidazole, CDTA(-1, -2), Na$_2$CO$_3$(-1, -2), KOH(0.5, 1.0 and 4M), to leave a residue. These were analysed for their carbohydrate and phenolic acids composition. Arabinose and galactose were the main noncellulosic sugars. Phenolics esterified to cell walls in carrot were found to consist primarily of p-hydroxybenzoic acid with minor contribution from vanillin, ferulic acid and p-hydroxybenzaldehyde. p-Hydroxybenzoic acid was quite strongly bound to the cell wall. The contents of p-hydroxybenzoic acid in 0.5M KOH, Na$_2$CO$_3$-2, IM KOH, and ${\alpha}$-cellulose were 2,097, 1,360, 1,140, and 717 $\mu\textrm{g}$/g AIR from CWM, respectively. Alkali labile unknown aromatic compound(C$\sub$7/H$\sub$10/O$_2$) was found in ${\alpha}$ -cellulose hydrolyzate digested with driselase and cellulase. This compound was also found in hydrolyzate of 2 M trifluoroacetic acid at 120$^{\circ}C$ for 2 hours. Driselase treatment solubilized only 46.6 $\mu\textrm{g}$/g of the p-hydroxybenzoic acid from carrot AIR. These results indicate that p-hydroxybenzoic acid was associated with neutral polysaccharides, long chain galactose and branched arabinan from graded alcohol precipitation.

• Food Science and Preservation
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• v.30 no.4
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• pp.573-588
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• 2023

The purpose of this study was to monitor changes in the quality of ginseng and predict its shelf-life. As the storage period of ginseng increased, some quality indicators, such as water-soluble pectin (WSP), CDTA-soluble pectin (CSP), cellulose, weight loss, and microbial growth increased, while others (Na₂CO₃-soluble pectin/NSP, hemicellulose, starch, and firmness) decreased. Principal component analysis (PCA) was performed using the quality attribute data and the principal component 1 (PC1) scores extracted from the PCA results were applied to the multivariate analysis. The reaction rate at different temperatures and the temperature dependence of the reaction rate were determined using kinetic and Arrhenius models, respectively. Among the kinetic models, zeroth-order models with cellulose and a PC1 score provided an adequate fit for reaction rate estimation. Hence, the prediction model was constructed by applying the cellulose and PC1 scores to the zeroth-order kinetic and Arrhenius models. The prediction model with PC1 score showed higher R² values (0.877-0.919) than those of cellulose (0.797-0.863), indicating that multivariate analysis using PC1 score is more accurate for the shelf-life prediction of ginseng. The predicted shelf-life using the multivariate accelerated shelf-life test at 5, 20, and 35℃ was 40, 16, and 7 days, respectively.

• Analytical Science and Technology
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• v.9 no.3
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• pp.279-291
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• 1996

The new chelating resins, XAD-2, 4, 16-TAC and XAD-2, 4, 16-TAO were synthesized by Amberlite XAD-2, XAD-4, and XAD-16 macroreticular resins with 2-(2-thiazolylazo)-p-cresol(TAC) and 4-(2-thiazolylazo)orcinol(TAO) as functional groups and were characterized by elemental analysis and FT-IR spectrometry. It was found that the content of functional group in chelating resin was 0.60mmol/g in XAD-16-TAC and 0.68mmol/g in XAD-16-TAO respectively. The chelating resins were stable in acidic and alkaline solution and can be reused over 10 times. The sorption behavior of some metalions to two chelating resins was investigated by batch method, which included batch equilibrium, effect of pH, coexisting ions and masking agent. For the optimum condition of sorption, the time required for equilibrium was about 1 hour and optimum pH was 5. In the presence of anions such as ${SO_4}^{2-}$ and $CH_3COO^-$, the sorption of U(VI) ion was slightly reduced but other anions such as $Cl^-$ and $NO{_3}^-$ revealed no interference effect. Also, sorption capacity of U(VI) ion was decreased by addition of $CO{_3}^{2-}$ ion because of complex formation of $[UO_2(CO_3)_3]^{4-}$, but alkali metals and alkali earth metals including Na(I), K(I), Mg(II), and Ca(II) were not affected for the sorption extent. Masking agent, NTA showed better separation efficiency of U(VI) ion from coexisting metal ions such as Th(IV), Zr(IV), Hf(IV), Cu(II), Cd(II), Pb(II), Ni(II), Zn(II) and Mn(II) than EDTA, CDTA.