• Title/Summary/Keyword: Morphological Characterization

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Isolation from Chungkookjang and Characterization of a Bacterium Producing an Extracellular Protease of High Specific Activity (청국장으로부터 고 비활성 세포외 Protease 생산 세균의 분리 및 동정)

  • Park, Hee-Jin;Park, Heui-Dong
    • Food Science and Preservation
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    • v.17 no.3
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    • pp.410-417
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    • 2010
  • Several extracellular protease-producing bacteria were isolated from Chungkookjang, a traditional Korean food of fermented soybeans, on skim milk agar plates. Among these bacteria, strain D14 exhibited the highest production (15.2 U/mL) and specific activity (40.0 U/mg protein) of extracellular protease activity as assessed on growth in a protease induction medium composed of 1% (w/v) soluble starch, 1.5% (w/v) skim milk, 0.5% (w/v) yeast extract, and 2% (w/v) NaCl. The bacterium was identified as Bacillus subtilis based on morphological and physiological characteristics and 16S rDNA sequence. A BLAST search of 16S rDNA sequences revealed that the isolate was most closely related to Bacillus subtilis subsp. subtilis strain NCIB 3610. The 16S rDNA sequence homology was 99.9%. Our isolate produced the highest level of protease when grown in a protease induction medium containing 1% (w/v) sorbitol and 0.5% (w/v) yeast extract. Fructose and glucose reduced enzyme production to 12.7% and 35.9%, respectively, of the level seen when the strain was grown in medium containing soluble starch. Soytone also reduced enzyme production to 61.4% of the level noted when the strain was grown in medium containing yeast extract.

Microbial Diversity of the Trichloroethylene Contaminated Groundwater Treatment System and Characterization of Pseudomonas sp. DHC8 (Trichloroethylene으로 오염된 지하수 제거공정의 미생물 다양성 및 분리균주 Pseudomonas sp. DHC8의 특성)

  • Nam, Ji-Hyun;Shin, Ji-Hye;Kwon, Kiwook;Bae, Wookeun;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.49 no.4
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    • pp.336-342
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    • 2013
  • Trichloroethylene (TCE) is a widely used substance in commercial and industrial applications, yet it must be removed from the contaminated soil and groundwater environment due to its toxic and carcinogenic nature. We investigated bacterial community structure, dominant bacterial strain, and removal efficiency in a TCE contaminated groundwater treatment system using immobilized carrier. The microbial diversity was determined by the nucleotide sequences of 16S rRNA gene library. The major bacterial population of the contaminated groundwater treatment system was belonging to BTEX degradation bacteria. The bacterial community consisted mainly of one genus of Pseudomonas (Pseudomonas putida group). The domination of Pseudomonas putida group may be caused by high concentration of toluene and TCE. Furthermore, we isolated a toluene and TCE degrading bacterium, named Pseudomonas sp. DHC8, from the immobilized carrier in bioreactor which was designed to remove TCE from the contaminated ground water. Based on the results of morphological and physiological characteristics, and 16S rRNA gene sequence analysis, strain DHC8 was identified as a member of Pseudomonas putida group. When TCE (0.83 mg/L) and toluene (60.61 mg/L) were degraded by this strain, removal efficiencies were 72.3% and 100% for 12.5 h, respectively. Toluene removal rate was 2.89 ${\mu}mol/g$-DCW/h and TCE removal rate was 0.02 ${\mu}mol/g$-DCW/h. These findings will be helpful for maintaining maximum TCE removal efficiency of a reactor for bioremediation of TCE.

Characterization of Cellulolytic and Xylanolytic Enzymes of Bacillus licheniformis JK7 Isolated from the Rumen of a Native Korean Goat

  • Seo, J.K.;Park, T.S.;Kwon, I.H.;Piao, M.Y.;Lee, C.H.;Ha, Jong K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.1
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    • pp.50-58
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    • 2013
  • A facultative bacterium producing cellulolytic and hemicellulolytic enzymes was isolated from the rumen of a native Korean goat. The bacterium was identified as a Bacillus licheniformis on the basis of biochemical and morphological characteristics and 16S rDNA sequences, and has been designated Bacillus licheniformis JK7. Endoglucanase activities were higher than those of ${\beta}$-glucosidase and xylanase at all temperatures. Xylanase had the lowest activity among the three enzymes examined. The optimum temperature for the enzymes of Bacillus licheniformis JK7 was $70^{\circ}C$ for endoglucanase (0.75 U/ml) and $50^{\circ}C$ for ${\beta}$-glucosidase and xylanase (0.63 U/ml, 0.44 U/ml, respectively). All three enzymes were stable at a temperature range of 20 to $50^{\circ}C$. At $50^{\circ}C$, endoglucanse, ${\beta}$-glucosidase, and xylanase had 90.29, 94.80, and 88.69% residual activity, respectively. The optimal pH for the three enzymes was 5.0, at which their activity was 1.46, 1.10, and 1.08 U/ml, respectively. The activity of all three enzymes was stable in the pH range of 3.0 to 6.0. Endoglucanase activity was increased 113% by $K^+$, while $K^+$, $Zn^+$, and tween 20 enhanced ${\beta}$-glucosidase activity. Xylanase showed considerable activity even in presence of selected chemical additives, with the exception of $Mn^{2+}$ and $Cu^{2+}$. The broad range of optimum temperatures (20 to $40^{\circ}C$) and the stability under acidic pH (4 to 6) suggest that the cellulolytic enzymes of Bacillus licheniformis JK7 may be good candidates for use in the biofuel industry.

Characterization of Low-phosphorus Tolerance in an Anthocyanin-deficient Lycopersicon esculentum by tissue culture (조직 배양을 이용한 안토시아닌 결핍 돌연변이 토마토의 저인 내성 평가)

  • Lee, Dong-Hee;Bae, Gong-Young;Park, So-Hong
    • Korean Journal of Environmental Agriculture
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    • v.18 no.2
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    • pp.174-178
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    • 1999
  • An anthocyanin-deficient tomato (Lycopersicon esculentum Mill.) strain, H957, shows an unusual tolerance to low phosphorus (P). To investigate whether the tolerance originates from a tissue/cellular strength, plant tissue culture procedure was employed which facilitate to characterize the tolerance independent of morphological features. The tolerance was analyzed by comparing H957 against H883, its maternal wild type, while each explant was co-cultured on minimal P media. Comparisons were made in fresh weight, dry weight, callus and shoot formation, mineral contents, and P utilization ratios at $0-400{\cdot}\bar{I}MP$, . Growth of the two strains was severely impaired at 0 and $12.5{\cdot}\bar{I}MP.\;At\;25-200{\cdot}\bar{I}MP$, however, H957 consistently showed a greater fresh and dry weight than H883. Shoot onset of H957 was less delayed than H883 compared to optimal P conditions. H957 tissue contains an overall lower P concentration than H883. These observations indicate that H957 may tolerate to low P by its tissue or cellular strength in P utilization side from its morphology.

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Development and Characterization of a Hydrolyzed Goat Milk Protein/Chitosan Oligosaccharide Nano-Delivery System (산양유 단백질 분해물/키토올리고당 나노 전달체 제조 및 물리화학적 특성연구)

  • Ha, Ho-Kyung;Kim, Jin Wook;Han, Kyoung-Sik;Yun, Sung Seob;Lee, Mee-Ryung;Lee, Won-Jae
    • Journal of Dairy Science and Biotechnology
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    • v.35 no.3
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    • pp.208-214
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    • 2017
  • The aims of this study were to manufacture a hydrolyzed goat milk protein (HGMP)/chitosan ologisaccharide (CSO) nano-delivery system (NDS) and to investigate the effects of production variables, such as sodium tripolyphosphate (TPP), HGMP, and CSO concentration levels, on the formation and physicochemical properties of the NDS. An HGMP/CSO NDS was produced using the ionic gelation method at pH 5.5. Transmission electron microscopy and a particle size analyzer were used to determine the morphological and physicochemical properties of NDSs, respectively. The size of the HGMP/CSO NDS decreased from 225 to 138 nm as HGMP and CSO concentration levels decreased. The NDS had a positive surface charge, with a zeta-potential value of +23 mV. The encapsulation efficiency (EE) of docosahexaenoic acid was enhanced as the HGMP concentration level increased. Additionally, increasing the concentration level of CSO resulted in an increase in the EE of resveratrol. The HGMP/CSO NDS exhibited good physical stability during freeze-drying. Thus, our findings showed that the HGMP/CSO NDS was successfully manufactured and that HGMP and CSO concentration levels were key factors affecting the physicochemical properties of the NDS.

Characterization of PAH-Degrading Bacteria from Soils of Reed Rhizosphere in Sunchon Bay Using PAH Consortia (순천만 갈대근권 토양으로부터 얻은 PAH 분해세균의 특성 분석)

  • Kim Sung-Hyun;Kang Sung-Mi;Oh Kye-Heon;Kim Seung-Il;Yoon Byoung-Jun;Kahng Hyung-Yeel
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.208-215
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    • 2005
  • This study was accomplished in order to collect fundamental data on microbial roles in recycling process of reed rhizosphere. Sunchon bay, which is considered as one of the marsh and mud environments severely affected by human activities such agriculture and fisheries, was selected as a model place. In our initial efforts, two bacterial consortia were obtained by enrichment culture using PAH mixtures containing anthracene, naphthalene, phenanthrene and pyrene as the sources of carbon and energy, and four pure bacteria capable of rapid degradation of PAH were isolated from them. Four strains designated as SCB1, SCB2, SCB6, and SCB7 revealed by morphological, physiological and molecular analyses were identified as Burkholderia anthina, Alcaligenes sp., Achromobacter xylosoxidans., and Pseudomonas putida, respectively with over $99{\%}$ confidence. Notably, Burkholderia anthina SCB1 and Alcaligenes sp. SCB2 were found to utilize anthracene and pyrene more quickly than naphthalene and phenanthrene, whereas Achromobacter xylosoxidans SCB6 and Pseudomonas putida SCB7 exhibited similar growth and degradation patterns except for pyrene. These facts suggest that the rhizosphere microorganisms capable of PAH degradation might be used to clean up the contamination sites with polycyclic aromatic hydrocarbons.

Isolation and Characterization of Bacillus subtilis CH-10 Secreting Cellulase from Cattle Manure (우분으로부터 Bacillus subtilis CH-10의 분리 및 균주가 분비하는 Cellulase의 특성에 관한 연구)

  • Kim, Tae-Il;Han, Jung-Dae;Jeon, Byoung-Soo;Ha, Sang-Woo;Yang, Chang-Bum;Kim, Min-Kyun
    • Korean Journal of Microbiology
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    • v.35 no.4
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    • pp.277-282
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    • 1999
  • A bacterium producing the extracellular cellulase was isolated from cattle feces and screened as cellulase activity was excellent upon congo red straining method and activity measurements. Isolate was identified as Bacillus subtilis CH-10 on the basis of morphological and biochemical properties as well as cellular fatty acids composition. The enzyme which the isolate secretes had the optimum initial pH and temperature for its induction was 7.5 and 50${\circ}C$, respectively. The maximum CMCase activity in crude enzyme solution was observed at pH 7.5 and 75${\circ}C$ and was stable for pH 7.5 to 9.0 to maintain 70% activity. When the isolate was cultured in CMC media at 37${\circ}C$ for 24 hrs, CMCase and FPase activity was 1.13 U/㎖and 0.16U/㎖, respectively whereas Avicelase and ${\beta}$-glucosidase activity was not detected. When crude supernatant was used for zymogram, three major bands, cel 1, cel 2 and cel 3, were detected approximately 39, 41 and 57 KDa, respectively on CMC-SDS-PAGE.

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Isolation and Characterization of Bacteria Capable of Degrading Bisphenol A (Bisphenol A 분해세균의 분리 및 특성)

  • 김희식;이영기;이완석;박찬선;윤병대;오희목
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.189-196
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    • 2001
  • Eighty-seven microbial strains capable of growing on bisphenol A (BPA) as a sole carbon source were isolated from soils, waste waters and sludges. Among them, three bacterial strains were finally selected as potential decomposers through measuring BPA-degradation efficiency by HPLC analysis. Two of these bacterial strains were identified as Serratia marcescens 1901 and S. marcescens 1902, and another was Pseudomonas putida 1401 by 16S rDNA partial sequences and based on morphological and physiological properties. They showed higher cell growth and BPA degradation in PAV (PAS medium containing vitamin mixtures) than in PAS medium. The degradation efficiencies of these bacterial strains were within a range of 20-40% in the PAV containing 500 mg/1 or 100 mg/l of BPA fur 3 days. S. marcescens 1901 showed higher degradation efficiency at 100 mg/1 of BPA than those of other selected strains, while S. marcescens 1902 and P. putida 1401 degraded a high concentration of BPA (500 mg/l) with a degradation efficiency of 40% for 3 days. The BPA degradation using a mixed culture of three selected strains showed the similar level of dog-radation efficiency with that using a pure culture.

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Isolation and Characterization of a Novel Polysaccharide Producing Bacillus polymyxa A49 KCTC 4648P

  • Ahn, Sung-Gu;Suh, Hyun-Hyo;Lee, Chang-Ho;Moon, Seong-Hoon;Kim, Hee-Sik;Ahn, Keug-Hyun;Kwon, Gi-Seok;Oh, Hee-Mock;Yoon, Byung-Dae
    • Journal of Microbiology and Biotechnology
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    • v.8 no.2
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    • pp.171-177
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    • 1998
  • The strain A49, which produces a new type of extracellular polysaccharide was isolated from soil samples. From morphological, physiological and biochemical tests, the strain A49 was identified as a Bacillus polymyxa and named Bacillus polymyxa A49. Bacillus polymyxa A49 was found to produce a highly viscous extracellular polysaccharide when grown aerobically in a medium containing glucose as the sole source of carbon. The polysaccharide (A49 POL) showed a homogeneous pattern on gel permeation chromatography (GPC) and its molecular weight was estimated to be about 1.6 mega dalton (mDa). The FT-IR spectrum of A49-POL revealed typical characteristics of polysaccharides. As a result of investigations with HPLC and carbozole assay, A49-POL was found to consist of L-fucose, D-galactose, D-glucose, D-mannose, and D-glucuronic acid, with the molar ratio of these sugars being approximately 1:2:7:50:12. Rheological analysis of A49 POL revealed that it is pseudoplastic and has a higher apparent viscosity at dilute concentrations than does xanthan gum. The consistancy factor of A49 POL was found to be higher, and the flow index of A49 POL lower, than xanthan gum. Its apparent viscosity was comparatively unstable at various temperatures. the A49 POL showed the highest apparent viscosity at pH 3. When salts were added to A49 POL solution, the solution was compatible with up to 10% KCl, 35% NaCl, 55% $CaCl_2$, 55% $MgCl_2$, 55% $K_2HPO_4$, and 110% $Ca({NO_3})_2$, respectively.

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Characterization of Endochitosanases-Producing Bacillus cereus P16

  • Jo, Yu-Young;Jo, Kyu-Jong;Jin, Yu-Lan;Jung, Woo-Jin;Kuk, Ju-Hee;Kim, Kil-Yong;Kim, Tae-Hwan;Park, Ro-Dong
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.960-968
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    • 2003
  • A bacterial isolate showing a strong endochitosanase activity was isolated from soil and then characterized. The isolate was identified and designated as Bacillus cereus P16, based on morphological and biochemical properties, assimilation tests, cellular fatty acids pattern, along with 16S rRNA gene sequence. The optimized medium for producing extracellular chitosanase in a batch culture contained 1% tryptone, 0.5% chitosan, and 1% NaCl (pH 7.0). Powder chitosan and tryptone served the best as carbon and nitrogen sources, respectively, for the chitosanase production. Chitosanase activity was the highest when culture was completed at $37^{\circ}C$ among various temperatures ($20-42^{\circ}C$) tested in a shaking incubator (200 rpm). The levels of chitosanase activity in the culture fluid were 2.0 U/ml and 3.8 U/ml, respectively, when incubated in a flask for 60 h and in a jar fermenter for 24 h. The culture supernatant showed a strong liquefying activity on the soluble chitosan. The viscosity of 1% chitosan solution, that was incubated with the culture supernatant, was rapidly decreased, suggesting the secretion of endochitosanolytic enzymes by P16. The culture fluid revealed six endo-type chitosanase isozymes, two major (38 and 45 kD), and four minor (54, 65, 82, and 96 kD) forms by staining profile. The crude enzymes were very stable, and full activity was maintained for 4 weeks at $4^{\circ}C\;or\;-20^{\circ}C$ in the culture supernatant, suggesting a highly desirable stability rate for making an industrial application of the crude enzymes. The supernatant also cleaved the insoluble chitosan powder, but the hydrolysis rate was much lower. The enzymic degradation products of chitosan contained $(GlcN)_n$ (n=2-8). The concentration of chitosan in the reaction mixture of the crude enzyme affected the chitooligosaccharides composition of the hydrolysis products. When the higher concentration of chitosan was used, the higher degree of polymerized chitooligosaccharides were produced. By comparison with other commercial chitosanase preparations, P16 was indeed found to be a valuable enzyme source for industrial production of chitooligosaccharides from chitosan.