• Title/Summary/Keyword: biochemical characterization

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Isolation and Characterization of Bacillus Species Possessing Antifungal Activity against Ginseng Root Rot Pathogens (인삼 뿌리썩음병에 길항력이 있는 Bacillus 균의 분리 동정 및 특성 조사)

  • Kim, Byung-Yong;Ahn, Jae-Hyung;Weon, Hang-Yeon;Song, Jaekyeong;Kim, Sung-Il;Kim, Wan-Gyu
    • The Korean Journal of Pesticide Science
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    • v.16 no.4
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    • pp.357-363
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    • 2012
  • Ginseng (Panax ginseng C. A. Meyer) is an economically important crop in Korea. While the consumption of the crop is gradually increasing, the yield is decreasing due to the injury of continuous cultivation or infection of soil-borne fungal pathogens such as Cylindrocarpon destructans, Fusarium solani, Rhizoctonia solani and Sclerotinia nivalis. In order to find promising biocontrol agents, we have isolated 439 soil bacteria from ginseng cultivated soil and tested their antifungal activities against ginseng rot pathogens. Among them, 3 strains were finally selected and tested for the elucidation of their genetic and biochemical properties. They were identified as Bacillus amyloliquefaciens using phylogenetic analysis based on 16S rRNA gene sequences. Moreover, all selected strains showed positive reaction for PCR detection targeting biosynthetic gene sequences of iturin A and surfactin. The results provided promising evidences that the bacterial strains isolated from ginseng cultivated soil can be novel biocontrol agents for ginseng cultivaion.

Genetic characterization of Shigella flexneri isolated from the diarrheic patients in Seoul region (서울지역 설사환자로 부터 분리된 Shigella flexneri의 성상과 유전적 특성)

  • Seung, Hyun-Jung;Kim, Moo-Sang;Oh, Young-Hee;Choi, Byung-Hyun;Chae, Hee-Sun;Chu, Jiaqi;Jun, Moo-Hyung
    • Korean Journal of Veterinary Research
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    • v.46 no.4
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    • pp.337-345
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    • 2006
  • The shigellae are common etiological agents of bacillary dysentery in humans and primates. During four years from 2002 to 2005, 22 strains of Shigella spp. were isolated from the diarrheic patients in Seoul region. All of them were identified as S. flexneri by biochemical tests and serotyping. The prevalence of serotypes were variable by year, but the major serotypes were 2a and 3a. In an antimicrobial susceptibility test, all of the isolates were resistant to streptomycin and tetracycline, and susceptible to amikacin, kanamycin, cefoxitin, and gentamicin. All of the isolates showed the multi-resistant patterns over 3 drugs. By analysis of the plasmid profile the isolates were classified into 7 groups (P1~P7). Serotypes 2a and 2b were distributed to P1, P2, P3, and P4. Serotype 3a was differentiated to P5 and serotype 3b, to P6 and serotype 4a, to P7. PCR results showed that all isolates were positive for two virulence genes, ipaH and ial, but none of the strains had stx gene. The set1A and set1B genes were detected from 12 isolates (54.5%) that belonged to serotype 2a and 2b. The sen gene was detected from 19 isolates (86.4%). The 22 isolates showed 12 to 17 DNA fragments in the sizes ranging from 20.5 kb to 1135 kb, resulting in 13 patterns by the PFGE with Not I digestion. The PFGE patterns of the isolates showed the close relation with the serotypes, but no relations with year of isolation and antimicrobial resistance.

Characterization of Microorganisms in Eoyukjang (어육장의 미생물학적 특성)

  • Oh, Eu-Jin;Oh, Mi-Hwa;Lee, Jong-Mee;Cho, Mi-Sook;Oh, Sang-Suk
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.656-660
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    • 2008
  • Fermented soybean foods are an important component of the Korean diet. Eoyukjang is a type of traditional fermented soybean source. Microbial analysis of eoyukjang was conducted during the fermentation period in this study. Microorganisms isolated from eoyukjang were identified by biochemical tests and 16S rDNA sequencing. 17 different microorganisms, including bacteria, yeast, and fungi were detected in eoyukjang during the fermentation period. Even though Aspergillus participated in the early stage of fermentation of eoyukjang, Bacillus species and Saccharomyces cerevisiae were the major microzymes in eoyukjang throughout the maturation period. Eoyukjang is generally consumed after the boiling of the final sample. Therefore, the final sample of eoyukjang was boiled and analyzed. Our results showed that no vegetative microorganisms survived under the boiling conditions for eoyukjang. Fermented soybean products in the domestic market were also assessed for comparison with the results from eoyukjang. The total cell number of kanjang (soy sauce) samples was between 0 to 42 CFU/mL. The isolated microorganisms were identified as Bacillus species. All Bacillus isolates were not found to harbor the three enterotoxin-producing and emetic toxin-producing genes.

Characterization of proteases of Toxoplasma gondii (Toxoplasma gondii에서 단백질 분해 효소의 특징)

  • Choe, Won-Yeong;Nam, Ho-U;Yun, Ji-Hye
    • Parasites, Hosts and Diseases
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    • v.27 no.3
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    • pp.161-170
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    • 1989
  • The proteases of Toxoplasma gcndii were purified partially and characterisrd for some biochemical properties including various chromatographic patterns, major catalytic classes, and conditions to promote the activity of these enzymes. When Toxoplasma extract was incubated with 3H-casein at various pH, peak hydrolysis of casein was observed at pH 6.0 and at pH 8.5. Proteasfs working at pH 6.0 and at pH 8.5 were purified partially by conventional methods of chromatographies of DE52 anion rxchange, Sephadex G-200 gel permeation, and hydroxylapatite chromatography. Partially purified enzymes were tested by site-specific inhibitors and promotorf. The protease working at pH 6.0 was inactivated by iodoacetamide with LDso of 10-5 M and promoted by dithiothreitol, while the protease working at pH 8.5 was inhibited by phenylmethylsulfonyl fluoride with LD50 of 10-5 M and was Promoted by ATP (excess ATP beyond 2 mM inhibited the activity reversely). The protease of pH 8.5 had the activity of ATPase which might exert the energy to its action. Therefore the former was referred to as a cysteinyl acid protease and the latter, ATP-dependent neutral serine protease.

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Synthesis and Characterization of Interfacial Properties of a Cationic Surfactant Having Three Hydroxyl Groups (세 개의 히드록실기를 가진 양이온 계면활성제 합성 및 계면 특성에 관한 연구)

  • Lee, Byung Min;Kim, Ji-Hyun;Kim, Sung Soo;Lim, Jong Choo
    • Applied Chemistry for Engineering
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    • v.23 no.5
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    • pp.433-439
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    • 2012
  • In this study, a cationic surfactant BHMAS (N,N-bis-(3'-n-dodecyloxy-2'-hydroxypropyl)-N-methyl-2-hydroxyethylammonium methyl sulfate) having two lauryl and three hydroxyl groups was synthesized by the reaction of n-dodecyl glycidyl ether and 2-aminoethanol followed by the quarternization with dimethyl sulfate. The structure of the product was elucidated by $^{1}H-NMR$ and FT-IR. The CMC (critical micelle concentration) and surface tension of BHMAS at CMC condition were found to be $9.12\;{\times}\;10^{-4}$ mol/L and 28.71 mN/m respectively. Dynamic surface tension measurements using a maximum bubble pressure tensiometer indicated that a relatively long time was required to saturate the interface between air and aqueous surfactant solution. The interfacial tension measured between 1 wt% surfactant solution and n-decane reached an equilibrium value of 0.045 mN/m in 5 min. The adsorption capacity of the synthesized surfactant was observed to be excellent, which suggests that the surfactant can be used as a softening agent during a laundry process.

Purification and Characterization of Lipase from Acinetobacter sp. B2 Isolated from Oil­contaminated Soil (유류오염지역에서 분리한 Acinetobacter sp. B2로부터의 Lipase 정제 및 특성)

  • Son Seung Hwa;Park Kyeong Ryang
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.320-327
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    • 2004
  • Three hundreds thirty two bacterial colonies which were able to degrade crude oil were isolated from soil sam­ples that were contaminated with oil in Daejeon area. Among them, one bacterial strain was selected for this study based on its higher oil degrading ability, and this selected bacterial strain was identified as Acinetobactor sp. B2 through physiological-biochemical tests and analysis of its 16S rRNA sequence. Acinetobactor sp. B2 was able to utilize various carbohydrates but did not utilize trehalose and mannitol as a sole carbon source. Acinetobactor sp. B2 showed a weak resistance to antibiotics such as kanamycin, streptomycin, tetracycline and spectinomycin, but showed a high resistance up to mg/ml unit to heavy metals such as Ba, Li, Mn, AI, Cr and Pb. The optimal growth temperature of Acinetobactor sp. B2 was $30^{\circ}C.$ The lipase produced by Acinetobactor sp. B2 was purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Its molecular mass was about 60 kDa and condition for the optimal activity was observed at $40^{\circ}C$ and pH 10, respectively. The activation energy of lipase for the hydrolysis of p­nitrophenyl palmitate was 2.7 kcal/mol in the temperature range of 4 to $37^{\circ}C,$ and the enzyme was unstable at the temperature higher than $60^{\circ}C.$ The Michaelis constant $(K_m)\;and\;V_{max}$ for p-nitrophenyl palmitate were 21.8 uM and $270.3\;{\mu}M\;min^{-1}mg^{-1},$ respectively. This enzyme was strongly inhibited by 10 mM $Cd^{2+},\;Co^{2+},\;Fe^{2+},\;Hg^{2+},$ EDTA and 2-Mercaptoethalol.

Biochemical Characterization of an Extracellular Protease in Serratia proteamaculans Isolated from a Spider (무당거미에서 분리한 Serratia proteamaculans에서 분비되는 단백질분해효소의 생화학적 특성)

  • Lee Kieun;Kim Chul-Hee;Kwon Hyun-Jung;Kwak Jangyul;Shin Dong-Ha;Park Doo-Sang;Bae Kyung-Sook;Park Ho-Yong
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.269-274
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    • 2004
  • Serratia proteamaculans isolated from the midgut of a spider formed big halos around the bacterial colonies, indicating that the bacterial strain produces an extracellular protease. Activity staining of the extracellular pro­tein fractions using zymogram also demonstrated that the major protein with an estimated molecular mass of 52 kDa contained a high proteolytic activity. The protease was purified to near electrophoretic homogeneity from the culture supernatant after filtration and ion exchange and size exclusion chromatography. The purified enzyme had a relatively high proteolytic activity between pH 6.0 and 10.0 and at broad temperature range. The proteolytic activity of the enzyme was not inhibited by phenylmethylsulfonyl fluoride but strongly inhibited by 1, 10-phenanthroline and EDTA. The activity also was dependent on the presence of $Ca^{++}\;and\;Zn^{++}$ ions. These observations indicate that the enzyme is a metalloprotease.

Isolation of Bacillus sp. Producing Poly-$\gamma$-glutamic Acid with High Efficiency and Its Characterization (고효율 Poly-$\gamma$-Glutamic Acid생산 균주의 분리 및 생산 특성)

  • You Kyung-Ok;Oh You-Na;Kim Byung-Woo;Nam Soo-Wan;Jeon Sung-Jong;Kim Dong-Eun;Kim Young-Man;Kwon Hyun-Ju
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.200-206
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    • 2005
  • A bacterium with high productivity of poly-$\gamma$-glutamic acid (PGA) was isolated from the traditional Korean seasoning, ChungKookJang. The 16s ribosomal RNA sequence of isolated strain showed 97.6, 98.9 and $90.3{\%}$ of similarity to those of Bacillus sp. WL-3, Bacillus subtilis; ENV1 and B amy-loliquefaciens (T), respectively. Accordingly, this bacterium was identified as a Bacillus sp. However, some biochemical characteristics of this strain were different from those of B. subtilis: D-xylose fermentation and glycogen utility were negative. Maximum production of PGA was achieved when it was grown aerobically in a culture medium containing glutamic acid ($3{\%}$) and fructose ($4{\%}$) as carbon sources. The volumetric yield of PGA reached up to 27 g/l in the optimum culture medium. These results suggest that the present strain can be applicable for industrial purposes such as a prototype strain for food or cosmetics industry.

Isolation and Characterization of Oligotrophic Strains with High Enzyme Activity from Buckwheat Sokseongjang (메밀 속성장 유래 효소활성 우수 저영양성 균주 분리 및 특성)

  • Lee, Sung-Young;Kim, Ji-Yeun;Baek, Sung-Yeol;Yeo, Soo-Hwan;Koo, Bon-Sung;Park, Hye-Young;Choi, Hye-Sun
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.735-741
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    • 2011
  • Bealmijang is a short-term fermented regional product that is prepared with soybean and extra ingredients. In this study, starter strain candidates were screened from Bealmijang for fermented soybean paste products. Twenty one bacterial strains producing extracellular enzymes (amylase, cellulase, protease, xylanase and lipase) were isolated from Bealmijang, buckwheat sokseongjang. The isolates were assessed for fibrinolytic and antibacterial activities, and salt tolerance. Strain HJ18-4, identified as Bacillus subtilis (AB601598) by biochemical properties (89.6%) and 16S rDNA sequencing (100%), showed the highest enzymatic, fibrinolytic, and antibacterial activities among the isolates. Although the growth of HJ18-4 was inhibited by the increase of NaCl concentration, the growth still exceeded that of B. subtilis KACC 10114 at 5% and 10% NaCl. These results suggest that B. subtilis HJ18-4 is suitable as a starter for soybean paste manufacture.

Biological characterization of Tenacibaculum maritimum isolated from cultured olive flounder in Korea and sensitivity against native plant extracts (한국의 양식넙치에서 분리한 Tenacibaculum maritimum의 특성과 자생식물 추출물에 대한 감수성)

  • Jang, Yeoung-Hwan;Jeong, Joon-Bum;Yeo, In-Kyu;Kim, Ki-Young;Harikrishnan, Ramasamy;Heo, Moon-Soo
    • Journal of fish pathology
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    • v.22 no.1
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    • pp.53-65
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    • 2009
  • Tenacibaculum maritimum (formerly Flexibacter maritimus) is the aetiological agent of an ulcerative and necrotic disease commonly called tenacibaculosis in marine fish. Tenacibaculosis is an economically important disease in a great variety species in Jeju Island cultured fish and leading to this pathogen initially affected by skin, mouth, fins, tail causing severe necrotic and ulcerative lesions on the body surface. In the present study, A-7 strain was isolated from Paralichthys olivaceus showing symptoms of tenacibaculosis and identified as T. maritimum by morphological, biochemical and molecular biological analysis. T. maritimum A-7 is experimentally infected through immersion route in Paralichthys olivaceus which the disease outbreaks in land-based fish tanks of Jeju Island. Up to data a number of treatments proposed for the tenacibaculosis outbreaks are based on the immersion administration of drugs in tank. Oxytetracycline is the most widely used disinfectants in fish farms. However, most of fish farms manager and consumers have expressed concern as bioaccumulation in tissue and its environmental. In addition, this antimicrobial compounds is expensive in fish farmers. The overcome of this problem is desired the application of natural plant derived products. To obtain as 70% EtOH extract antimicrobial compounds against tenacibaculosis from 35 species of Jeju Island native plants were screened for antimicrobial activity against T. maritimum. In the present study were identified most of the plant extracts were better antimicrobial activity against T. maritimum.