• Title/Summary/Keyword: Soil enzyme activity

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Field Gas-Sparging Tests for In Situ Aerobic Cometabolism of Trichloroethylene(TCE)

  • Kim Young;Istok Jonathan D.;Semprini Lewis;Oa Sung-Wook
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2006.04a
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    • pp.54-56
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    • 2006
  • Single-well-gas-sparging tests were developed and evaluated for assessing the feasibility of in-situ aerobic cometabolism of trichloroethylene (TCE), using propane as a growth substrate. To evaluate transport characteristics of dissolved solutes [sulfur hexafluoride (SF6) or bromide (non-reactive tracers), propane (a growth substrate), ethylene, propylene (nontoxic surrogates to probe for CAH transformation activity), and DO], push-pull transport tests were performed. Mass balance showed about 90% of the injected bromide and about 80% of the injected SF6 were recovered, and the recoveries of other solutes were comparable with bromide and slightly higher than SF6. A series of Gas-Sparging Biostimulation tests were performed by sparging propane/oxygen/argon/SF6 gas mixtures, and temporal ground water samples were obtained from the injection well under natural gradient 'drift' conditions. The decreased time for propane depletion and the longer time to deplete SF6 as a conservative tracer indicate the progress of biostimulation. Gas-Sparging Activity tests were performed. .Propane utilization, DO consumption, and ethylene and propylene cometabolism were well demonstrated. The stimulated propane-utilizers cometabolized ethylene and propylene to produce ethylene oxide and propylene oxide, as cometabolic by-products, respectively. Gas-Sparging Acetylene Blocking tests were performed by sparging gas mixtures including acetylene to demonstrate the involvement of monooxygenase enzymes. Gas substrate degradation was essentially completely Inhibited in the presence of acetylene, and no production of the corresponding oxides was also observed. The Gas-Sparging tests supports the evidences that the successive stimulation of propane-oxidizing microorganisms, cometabolic transformation of ethylene and propylene by the enzyme responsible for methane and propane degradation.

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Dynamics of Soil Microflora and Soil Enzymes around the Fairy-rings of Tricholoma matsutake (송이 균환(菌環) 주변(周邊)의 토양미생물(土壤微生物)과 토양효소(土壤酵素)의 동태(動態))

  • Hur, Tae Chul;Park, Hyun
    • Journal of Korean Society of Forest Science
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    • v.90 no.6
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    • pp.767-773
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    • 2001
  • We investigated the impacts of the fairy-ring of Tricholoma matsutake on the dynamics of soil microflora and soil enzyme activities by grouping the soils around the fairy-ring of T. matsutake into four regions. The regions were grouped as 'zone of decayed mycorrhizae', 'zone of mycorrhizae for fruiting', 'zone of physiologically active mycorrhizae' and 'zone free from mycorrhizal infection'. Soil fungi and actinomycetes were quite little at the soils around the fairy-ring of T. matsutake compared to those of general forest soils, and there were significant differences among the four regions. The soils with the mycelial cluster of T. matsutake showed about one third of microbial population compared to those in the zone free from mycorrhizal infection, which indicated that T. matsutake took a dominant position within the fairy-ring of the fungus. We could manifest that T. matsutake showed a distinctive characteristics of mycorrhizal fungus since the activities of dehydrogenase were significantly different between the zone of physiologically active mycorrhizae and the zone free from mycorrhizal infection. The dehydrogenase activity was the highest at the early season of fruiting around the fairy-ring of T. matsutake, while the acid-phosphatase activity increased from March to June followed by a slight decrease on August and peaked on October. This phenomenon made us infer that the vitality of T. matsutake be sustained after fruiting.

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Comparison of specific activity and cytopathic effects of purified 33 kDa serine proteinase from Acanthamoeba strains with different degree of virulence

  • Kim, Won-Tae;Kong, Hyun-Hee;Ha, Young-Ran;Hong, Yeon-Chul;Jeong, Hae-Jin;Yu, Hak-Sun;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.44 no.4 s.140
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    • pp.321-330
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    • 2006
  • The pathogenic mechanism of granulomatous amebic encephalitis (GAE) and amebic keratitis (AK) by Acanthamoeba has yet to be clarified. Pretense has been recognized to play an important role in the pathogenesis of GAE and AK. In the present study, we have compared specific activity and cytopathic effects (CPE) of purified 33 kDa serine proteinases from Acanthamoeba strains with different degree of virulence (A. healyi OC-3A, A. lugdunensis KA/E2, and A. castelianii Neff). Trophozoites of the 3 strains revealed different degrees of CPE on human corneal epithelial (HCE) cells. The effect was remarkably reduced by adding phenylmethylsulfonylfluoride (PMSF), a serine proteinase inhibitor. This result indicated that PMSF-susceptible proteinase is the main component causing cytopathy to HCE cells by Acanthamoeba. The purified 33 kDa serine proteinase showed strong activity toward HCE cells and extracellular matrix proteins. The purified proteinase from OC-3A, the most virulent strain, demonstrated the highest enzyme activity compared to KA/E2, an ocular isolate, and Neff, a soil isolate. Polyclonal antibodies against the purified 33 kDa serine proteinase inhibit almost completely the proteolytic activity of culture supernatant of Acanthamoeba. In line with these results, the 33 kDa serine proteinase is suggested to play an important role in pathogenesis and to be the main component of virulence factor of Acanthamoeba.

Biocontrol of Damping-Off(Rhizoctonia solani) in Cucumber by Trichoderma asperellum T-5 (Trichoderma asperellum T-5를 이용한 오이 모잘록병(Rhizoctonia solani)의 생물학적 제어)

  • Ryu, Ji-Yeon;Jin, Rong-De;Kim, Yong-Woong;Lee, Hyang-Burm;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.4
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    • pp.185-194
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    • 2006
  • A fungal strain of Trichoderma having strong chitinolytic activity was isolated from field soil enriched with crabshell for several years. Based on 5.8S rRNA, partial 18S, 28S rRNA genes, ITS1, ITS2 sequence analysis and morphological characteristics, the fungus was identified as Trichoderma asperellum and named as Trichoderma asperellum T-5 (TaT-5). The fungus released lytic enzymes such as chitinase and ${\beta}$-1, 3-glucanse, and produced six antifungal substances in chitin broth medium. To demonstrate the protective effect of TaT-5 against damping-off in cucumber plant caused by Rhizoctonia solani, TaT-5 culture broth (TA), chitin medium (CM) and distilled water (DW) were applied to each pot at 10 days after sowing, respectively. Then, the homogenized hyphae of R. solani were infected to each pot at 1 week after TaT-5 inoculation. During experimental period, fresh weight of shoot and root in cucumber plant more increased at TA treatment compared to other treatments. PR-proteins (${\beta}$-1, 3-glucanase and chitinase) activities in cucumber leaves markedly increased at CM and DW treatments, but the activity slightly increased and then decreased at TA treatment at 3 days after infection of R. solani. The activity of PR-proteins activities in cucumber roots at all treatments decreased with time where the degree of decrement was more alleviated at TA treatment than CM and DW. These results suggest that the lytic enzymes (chitinase and ${\beta}$-1, 3-glucanse) and antifungal substances produced by TaT-5 can reduce the pathogenic attack by R. solani in cucumber plants.

Identification and Biochemical Characterization of a New Xylan-degrading Streptomyces atrovirens Subspecies WJ-2 Isolated from Soil of Jeju Island in Korea (제주도 토양으로부터 자일란 분해 Streptomyces atrovirens subspecies WJ-2 동정 및 효소의 생화학적 특성 규명)

  • Kim, Da Som;Bae, Chang Hwan;Yeo, Joo Hong;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.512-521
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    • 2016
  • A bacterial strain was isolated from a soil sample collected on Jeju Island, Korea. The strain, designated WJ-2, exhibited a high xylanase activity, whereas cellulase activity was not detected. The 16S rRNA gene sequence of WJ-2 was highly similar to type strains of the genus Streptomyces. A neighbor-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain WJ-2 is phylogenetically related to Streptomyces atrovirens. Furthermore, DNA-DNA hybridization analysis confirmed that strain WJ-2 is a novel subspecies of Streptomyces atrovirens. The genomic DNA G+C content was 73.98 mol% and the major fatty acid present was anteiso-C15:0 (36.19%). The growth and xylanase production of strain WJ-2 were significantly enhanced by using soytone and xylan as nitrogen and carbon sources, respectively. Crude enzyme preparations from the culture broth of strain WJ-2 exhibited maximal total xylanase activities at pH 7.0 and $55^{\circ}C$. Thin-layer chromatography analysis revealed that the crude enzyme degrades beechwood xylan to yield xylobiose and xylotriose as the principal hydrolyzed end products.

Soil properties of barrier island habitats in the Nakdong river estuary (낙동강 하구 주요 사주 서식지 토양 특성)

  • Yi, Yong Min;Yeo, Un Sang;Sung, Kijune
    • Journal of Wetlands Research
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    • v.16 no.3
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    • pp.355-362
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    • 2014
  • Changes of soil properties due to sedimentation and erosion in the river estuary may lead changes in environmental factors that affect plant growth and distribution, Then habitats in the river estuary that provide various ecological functions can also be influenced. Topsoil samples were analyzed in order to understand the soil properties of important barrier islands and habitat types in the Nakdong river estuary. The samples were obtained from Phragmites communis and Scirpus planiculmis habitats, the tidal flats in the southern area of Eulsukdo, and in Mangeummerydeung, Baekhapdeung, and Doyodeung. Analyses results showed that bulk density, pH, organic matter content and total nitrogen concentration which were directly or indirectly affected by vegetation showed significant difference (p<0.05) with habitat types but no differences in water content and oxidation reduction potential which could be affected by soil texture and showed significant difference among barrier islands. Results suggested that soil properties on barrier islands in the Nakdong river estuary were influenced first by geomorphic changes due to sedimentation and erosion, and then by the presence or type of vegetation. A range of physical and chemical properties were analyzed; soil water content and bulk density (physical properties), and organic content and pH (chemical properties) were correlated with seven other soil properties, at a level of significance higher than 90%. These aspects played an important role in determining overall soil properties in the studied area.

Changes of Biological and Chemical Properties during Composting of Livestock Manure with Isolated Native Microbe (토착미생물별 가축분 퇴비화 과정중 생물화학적 특성 변화)

  • Han, Hyo-Shim;Lee, Kyung-Dong
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.6
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    • pp.1126-1135
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    • 2012
  • In order to produce high-quality fermenting composts, bacteria strains with high activities of extracellular enzymes (cellulase, chitinase, amylase, protease and lipase) were isolated from the soils in 6 provinces of Korea, and characterized by 16S rRNA gene sequence analysis and properties. The selected 7 stains inoculated to livestock manure for 2' fermenting time, and experimental treatment divided into 3 groups, B1, B2 and B3, according to microbial activity and enzyme type. Our results showed that microbe applications (B1, B2 and B3) can increase (p<0.05) both rhizomes (17-38%) and enzyme activities (50-81%) in compost after fermenting time, respectively, compared to non-microbe treatment (control). The microbe application also decreased significantly (p<0.05) the $NH_3$ and $H_2S$ gas contents 13.4 and 27.3% compared with control, and the Propionic acid and Butyric acid gas contents 14.5 and 19.6%, respectively, as compared to the control. The microbial degradation rate (%) of pesticides and heavy metals increased significantly (p<0.05) after fermenting time, respectively, as compared to the control. Especially, microbe applications were more effective in total rhizomes yields and bioactivities than non-microbe treatment. Thus the results of this study could help in development of potential bioinoculants and composting techniques that maybe suitable for crop production, and protectable for earth environment under various conditions.

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.

Isolation, Identification and Enzymatic Activity of Halotolerant and Halophilic Fungi from the Great Sebkha of Oran in Northwestern of Algeria

  • Chamekh, Rajaa;Deniel, Franck;Donot, Christelle;Jany, Jean-Luc;Nodet, Patrice;Belabid, Lakhder
    • Mycobiology
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    • v.47 no.2
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    • pp.230-241
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    • 2019
  • The Great Sebkha of Oran is a closed depression located in northwestern of Algeria. Despite the ranking of this sebkha among the wetlands of global importance by Ramsar Convention in 2002, no studies on the fungal community in this area have been carried out. In our study, samples were collected from two different regions. The first region is characterized by halophilic vegetation and cereal crops and the second by a total absence of vegetation. The isolated strains were identified morphologically then by molecular analysis. The biotechnological interest of the strains was evaluated by testing their ability to grow at different concentration of NaCl and to produce extracellular enzymes (i.e., lipase, amylase, protease, and cellulase) on solid medium. The results showed that the soil of sebkha is alkaline, with the exception of the soil of cereal crops that is neutral, and extremely saline. In this work, the species Gymnoascus halophilus, Trichoderma gamsii, the two phytopathogenic fungi, Fusarium brachygibbosum and Penicillium allii, and the teleomorphic form of P. longicatenatum observed for the first time in this species, were isolated for the first time in Algeria. The halotolerance test revealed that the majority of the isolated are halotolerant. Wallemia sp. and two strains of G. halophilus are the only obligate halophilic strains. All strains are capable to secrete at least one of the four tested enzymes. The most interesting species presenting the highest enzymatic index were Aspergillus sp. strain A4, Chaetomium sp. strain H1, P. vinaceum, G. halophilus, Wallemia sp. and Ustilago cynodontis.

Characterization of the \beta-Galactosidase Produced by Streptomyces sp. YB-10 (\beta-Galactosidase를 생산하는 Streptomyces sp. YB-10의 분리 및 효소 특성)

  • 윤기홍;이경섭;김창진
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.151-156
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    • 2003
  • A strain YB-10 was isolated from soil as a producer of the extracellular $\beta$-D-galactosidase, which catalyzes the hydrolysis of lactose. The strain YB-10 was identified as Streptomyces sp. on the basis of its cultural, morphological and physiological properties. After treating culture supernatant of the isolate with ammonium sulfate, the precipitated protein was used as a crude $\beta$-galactosidase for analyzing its reaction properties with para-nitrophenyl-$\beta$-D-galactosidase(pNP-$\beta$Gal) as a substrate. The $\beta$-galactosidase showed its maximal activity at pH 6.0 and 6$0^{\circ}C$. The enzyme was also active on lactose. The hydrolyzing activity of $\beta$-galactosldase for pNP-$\beta$Gal and lactose was decreased by galactose. Its hydrolyzing activity far lactose was also decreased by glucose, but the activity for pNP-$\beta$Gal was increased to 1.8-folds by glucose.