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Biochemical Characterization of an Extracellular Xylanase from Aestuariibacter sp. PX-1 Newly Isolated from the Coastal Seawater of Jeju Island in Korea (대한민국 제주도 연안 해수에서 새롭게 분리한 Aestuariibacter sp. PX-1이 생산하는 자일라네이즈의 생화학적 특성)

  • Kim, Jong-Hee
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.215-222
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    • 2020
  • The marine microorganism PX-1, which can hydrolyze xylan, was isolated from coastal sea water of Jeju Island, Korea. Based on the 16S rRNA gene sequence and chemotaxonomy analysis, PX-1 was identified as a species of the genus Aestuariibacter and named Aestuariibacter sp PX-1. From the culture broth of PX-1, an extracellular xylanase was purified to homogeneity through ammonium sulfate precipitation and subsequent adsorption chromatography using insoluble xylan. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography estimated the molecular weight of the purified putative xylanase (XylA) as approximately 64 kDa. XylA showed xylanase activity toward beechwood xylan, with a maximum enzymatic activity at pH 6.0 and 45℃. Through thin-layer chromatographic analysis of the xylan hydrolysate produced by XylA, it was confirmed that XylA is an endo-type xylanase that decomposes xylan into xylose and xyloligosaccharides of various lengths. The Km and Vmax values of XylA for beechwood xylan were 27.78 mM and 78.13 μM/min, respectively.

Effects of Loess on the Mycellial Pellet Formation of Phosphate-solubilizing Fungus, Aspergillus sp. PS-104 in the Submerged Culture (Aspergillus sp. PS-104의 액침배양증 mycellial pellet 형성에 미치는 황토의 영향)

  • Kang, Sun-Chul;Koo, Bon-Sung;Tae, Un-Hee
    • Korean Journal of Environmental Agriculture
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    • v.21 no.1
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    • pp.17-23
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    • 2002
  • In order to investigate effects if loess on the mycellial pellet formation a phosphate-solubilizing fungus. Aspergillus sp. PS-104 was cultured in potato dextrose broth containing loess. The strain formed an amorphous pellet or loose aggregates agitated at a low speed (50 rpm) while spherical and regular pellets at a high speed (150 rpm) The higher concentration of loess was added, the smaller size of a pellet was formed during the submerged culture of the strain. As shown in results, being cultured in the PDB medium supplemented with 1.0% loess the pellet size was maximally reduced to a fourth compared to the control. Evaluating the addition effect of several components of loess such as $SiO_2$, $Fe_2O_3$, $Al_2O_3$, $CaCO_3$, $CaSO_4$ and $MgCO_3$ on the reduction of mycellial pellet size the higher concentration was supplied, the smaller size of pellet was formed except $Al_2O_3$. And the smallest pellet size was recorded at the concentration of 1.0% (W/V) magnesium carbonate.

Characteristics and Action Pattern of ${\alpha}-galactosidase$ from Scopulariopsis brevicaulis in Korean Traditional Meju (한국 재래 간장에서 분리한 Scopulariopsis brevicaulis가 생성하는 ${\alpha}-galactosidase$의 특성 및 작용양상)

  • Choi, Kwang-Soo;Lee, Seon-Ho;Hong, Seung-Pyo;Lee, Hee-Duck;Bae, Du-Kyung;Choi, Cheong
    • Applied Biological Chemistry
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    • v.41 no.7
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    • pp.489-495
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    • 1998
  • The optimum culture condition of Scopulariopsis brevicaulis for the production of ${\alpha}$-galactosidase was as follows: Tryptone 1.5%, $NH_4NO_3$ 0.2%, Raffinose 2.5%, $KH_2PO_4$ 0.5%, yeast extract 0.5%, pH 7.0, $27^{\circ}C$. The optimum pH and temperature for the enzyme activity of ${\alpha}$-galactosidase producing Scopulariopsis brevicaulis were pH 7.0 and $27^{\circ}C$, respectively. The enzyme was relatively stable at $pH\;6.0{\sim}8.0$ and at temperature below $40^{\circ}C$. The activity of the enzyme was inhibited by $Ag^{2+},\;Hg^{2+},\;Cu^{2+}$, p-chloromercuribenzoic acid and Iodine. These results would indicate the presence of -SH groups in the catalytic site of the enzyme. Km value was 1.9 mM for $p-nitrophenyl-{\alpha}-D-galactopyranoside$ and Vmax value was $9.66{\times}10^2\;{\mu}M/min$. Sugar constituents of culture broth were identified by HPLC that the enzyme liberated sucrose, glucose and fructose from raffinose and raffinose was significantly decreased.

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Purification and Properties of the Polyvinyl alcohol oxidase from Xanthomonas campestris J2Y (폴리비닐 알콜 분해균 Xanthomonas campestris J2Y의 Polyvinyl alcohol oxidase 정제 및 성질)

  • Kwoen, Dae-Jun;Jo, Youl-Lae
    • Applied Biological Chemistry
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    • v.39 no.5
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    • pp.349-354
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    • 1996
  • The Polyvinyl alcohol(PVA) oxidase involved in PVA degradation by microorganism has been purified to homogeneity from culture broth of Xanthomonas campestris J2Y grown in a minimal medium containing PVA as a sole carbon source. The enzyme was purified by DEAE-cellulose chromatograpy and Sephadex G-150 gel filtration. The purified PVA oxidase was electrophoretically homogeneous both in the absence and presence of SDS. The molecular weight of the enzyme was estimated to be about 55,000 daltons by SDS-polyacrylamide gel electrophoresis and Sephadex G-150 gel filtration. The native enzyme existed as a monomer. The optimal pH and temperature was shown to be pH 7 and $37^{\circ}C$ respectively. The activity of enzyme was stable below $55^{\circ}C$ and between pH range of $5{\sim}11$. The enzyme activity was significantly inhibited by metal compounds such as $Ag^{2+},\;Hg^{2+}$. While, metal ions such as $Mn^{2+},\;and\;Cu^{2+}$ stimulated the reaction. Km value of the enzyme for PVA was $7.04{\times}10^{-2}mmol/{\ell}$.

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Studies on the Glucose Isomerizing Enzyme -Part II. Some Properties of the Glucose Isomerizing Enzyme of K-17 strain- (포도당(葡萄糖) 이성화(異性化) 효소(酵素)에 관(關)한 연구(硏究) -제2보(第二報); 분리선정(分離選定)된 균주효소(菌株酵素)의 성질(性質)에 대(對)해서-)

  • Seu, J.H.;Kim, C.K.;Ki, W.K.;Rhee, I.K.;Kwon, T.J.;Woo, D.L.
    • Applied Biological Chemistry
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    • v.11
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    • pp.49-53
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    • 1969
  • Some properties of glucose isomerizing enzyme which produced by the strain K-17 in xylose containing nutrient broth medium were investigated. The optimal pH for enzyme reaction was indicated about 7.2 and optimal temperature was about $75^{\circ}C$. The same optimal temprature was indicated by both cell free extract and acetone dried cells using as enzyme. The glucose isomerizing enzyme from strain K-17 was not inhibited by the high concentration of substrate even in a suturated glucose solution, but most enzyme was inactivated by the heat treatment at $80^{\circ}C$. The maximum fructose forming ratio from glucose was about 50 percents.

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Characterization of Oil-Degradation Biosurfactant Produced by Bacillus sp. TBM40-3 (Bacillus sp. TBM40-3에 의해 생성된 Biosurfactant의 유류분해 특성)

  • Kim, Sun-Hee;Lee, Sang-Cheol;Yoo, Ju-Soon;Joo, Woo-Hong;Chung, Soo-Yeol;Choi, Yong-Lark
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.170-175
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    • 2004
  • In this paper we studied about Bacillus sp. TBM40-3 producing biosurfactants. The strains were isolated from Taeback Mountain soil and identified as Bacillus sp. by l6S rDNA nucleotides sequence analysis. The TBM40-3 was gram-positive and rod-shaped as observed by field emission scanning microscopy. After the cultivation TBM40-3 in LB broth for 90 h and the surface tension of supernatant was decreased to 29 mN/m. Emulsification activity and stability of crude biosurfactant was measured by using water-immiscible hydrocarbons and oil as substrate. Maximum emulsification activity and stability was obtained from soybean oil. Also, we confirmed that the TBM40-3 producing biosurfactant had an effect on crude oil while showing a superior effect as compared to chemically synthesized surfactants (SDS, Span85, Tween40, Triton X-100). As a result, the Bacillus sp. TBM40-3 producing biosurfactant had potent properties as an emulsifying agent and an emulsion stabilizing agent.

EVALUATION OF ENTEROCOCCUS FAECALIS REMOVAL EFFICACY OF THE ENDOVAC® AND ENDOACTIVATOR® INTRACANAL IRRIGATION METHODS (EndoVac®과 EndoActivator®를 이용한 근관세척법의 Enterococcus faecalis 제거 효율 평가)

  • Song, Seung-Gon;Park, Se-Hee;Cho, Kyung-Mo;Kim, Jin-Woo
    • Restorative Dentistry and Endodontics
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    • v.34 no.5
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    • pp.390-396
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    • 2009
  • The aim of this study was to evaluate endodontic irrigation methods with $EndoVac^{(R)}$ and $EndoActivator^{(R)}$ in the elimination of Enterococcus faecalis from the root canals. Extracted 70 human single-rooted teeth were used. The canals were instrumented by a crown-down technique with .04 taper ProFile to ISO size 40. After the teeth were autoclaved, the canals were inoculated with E. faecalis and incubated for 48 h. The teeth were randomly divided into three experimental groups of 20 teeth each according to canal irrigation methods and two control groups as follows: group 1 - $EndoVac^{(R)}$; group 2 - $EndoActivator^{(R)}$; group 3-Conventional needle irrigation method. After canal irrigation using 2.5% NaOCl. first samples (S1) were taken using sterile paper point. And the canals were filled with sterile brain heart infusion (BHI) broth and incubated for 24 h, then second samples (S2) were taken. The samples were cultured on BHI agar plate to determine the numbers of colony forming units (CFU). In first sampling (S1), only one canal of conventional method among the all experimental groups was positive cultured. In second sampling (S2), $EndoVac^{(R)}$ group showed the least positive culture numbers of E. faecalis. There was statistically significant difference between the $EndoVac^{(R)}$ and conventional needle irrigation methods in the mean value of Log CFU. According to the results of this study, $EndoVac^{(R)}$ showed better efficacy than conventional needle irrigation method in the elimination of E. faecalis from the root canal.

Antifungal Activity of Benzoic Acid from Bacillus subtilis GDYA-1 against Fungal Phytopathogens (Bacillus subtilis GDYA-1로부터 분리한 benzoic acid의 식물병원성 곰팡이에 대한 항균활성)

  • Yoon, Mi-Young;Seo, Kook-Hwa;Lee, Sang-Heon;Choi, Gyung-Ja;Jang, Kyoung-Soo;Choi, Yong-Ho;Cha, Byeong-Jin;Kim, Jin-Cheol
    • Research in Plant Disease
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    • v.18 no.2
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    • pp.109-116
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    • 2012
  • A bacterial strain antagonistic to some fungal phytopathogens was isolated from the stem of a Persimmon tree in Yeongam, Korea. This bacterium was identified as Bacillus subtilis by 16S rRNA gene sequencing and designated as B. subtilis GDYA-1. In in vivo experiment, the fermentation broth exhibited antifungal activities against Magnaporthe oryzae on rice plants, Phytophthora infestans on tomato plants, and Puccinia recondita on wheat plants. We isolated one antifungal compound and its chemical structure was determined by mass and $^1H$-NMR spectral data. The antifungal substance was identified as benzoic acid. It inhibited mycelial growth of M. oryzae, Rhizoctonia solani, Sclerotinia sclerotiorum, and P. capsici with minimum inhibition concentration (MIC) values, ranging from 62.5 to 125 ${\mu}g/ml$. Moreover, the substance effectively suppressed Phytophthora blight of red pepper caused by P. capsici in a pot experiment. To the author's knowledge, this is the first report on the antifungal activity of benzoic acid against phytopathogenic fungi. Benzoic acid and B. subtilis GDYA-1 may contribute to environmental-friendly protect crops from phytopathogenic fungi.

Application of a PCR Method for the Detection of Mycoplasma in Veterinary Live Viral Vaccines (동물용 생 바이러스 백신에서 Mycoplasma 검출을 위한 PCR 기법 적용)

  • Jeon Woo-Jin;Kim Byoung-Han;Jung Byeong-Yeal;An Dong-Jun;Yi Chul-Hyun;Jang Hwan;Chung Gab-Soo
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.269-274
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    • 2005
  • We evaluated the PCR assay and two commercialized PCR kits for the detection of mycoplasma in veterinary via live vaccines. The PCR assay could specifically detect all the tested Mycoplasma spp. and Acholeplasma spp., whereas two commercialized PCR kits did not. Also, the specificity of the PCR assay showed that 4 reference strains and 7 field isolates belonging to avian mycoplasma species could be all detected. The sensitivity of the PCR assay was determined using pure cultured Mycoplasma spp. and Acholeplasma spp. with a range of 1 to 100 colony forming units/ml in 9 CFR Mycoplasma broth. To test the availability of the PCR assay for veterinary live viral vaccines, A. laidlawii was artificially inoculated into the swine transmissible gastroenteritis-rota virus combined vaccine and canine parvovirus vaccine, respectively and the sensitivity of the PCR assay was similar with the result of cultured samples. In this study, the PCR assays could be used as rapid and sensitive methods for the detection of mycoplasma in veterinary live viral vaccines.

Isolation and Phylogenetic Characterization of Chitinase Producing Oligotrophic Bacteria (Chitinase생산 저영양세균의 분리 및 계통분류학적 특성)

  • Kim Soo-Jin;Kim Min-Young;Koo Bon-Sung;Yoon San-Hong;Yeo Yun-Soo;Park In-Cheol;Kim Yoon-Ji;Lee Jong-Wha;Whang Kyung-Sook
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.293-299
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    • 2005
  • Many isolates from soil of Korean ginseng rhizosphere did not show remarkable growth on full strength of the conventional nutrient broth (NB medium) but grew on its 100-fold dilution (DNB medium). Six hundred-forty strains were isolated as oligotrophic bacteria. In the course of screening for new bioactive compounds from oligotrophic bacteria from soil, 8 strains which had appeared to form of clear zone on a medium containing colloidal chitin as a sole carbon source were selected for further studies. Strain CR42 hydrolyzed a fluorogenic analogue of chitin, 4-methylumbelliferyl-D-glucosaminide (MUF-NAG) . Mo st of the culture supernatant of these isolates hydrolyzed 4-methylumbelliferyl-D-N,N'-diacetylchitobioside (MUF-diNAG). The isolates were heterogeneous and categorized to gamma- and beta-proteobacteria, Bacillaceae, Actinobactepia, and Bacteroides by 16S rRNA analysis. Two strains, WR164 and CR18, had a 16S rRNA sequence of $95-96\%$ identical to uncultured bacteria. It was observed that CR2 and CR75 could inhibit the growth of Colletotrichum gloeosporioides with hyphal extention-inhibition assay on PDA plate supplemented with $1\%$ colloidal chitin.