• 제목/요약/키워드: Extracellular enzyme activities

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Methylumbelliferyl 형광기질을 이용한 평판배지상의 미생물 체외 세포효소측정방법 (Microbial Extracellular Enzyme Detection on Agar Plates by Means of Fluorogenic Methylumbelliferyl-Substrates)

  • 김상진
    • 미생물학회지
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    • 제28권3호
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    • pp.229-235
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    • 1990
  • 평판배지상 세균 colony의 체외 세포 효소활성을 직접 측정할 수 있는 신속하고 정확한 방법에 대하여 기술하였다. 일반적으로 세균의 효소 특성을 살피기 위해서는 단백질, 전분, chitin, tween-80 등과 같은 고분자 물질을 첨가한 선택배지를 사용하고 있으나 그 방법상 여러 가지 문제점이 있다 그러므로 본 연구에서는 형광물질의 일종인 Methvlumbell liferyl(MUF) 기질이 일반적으보 사용되고 있는 천연 고분자 물질고 유사한가를 순수분리세균 균주를 이용하여 실험으로 검증하였다. MUF 기질 분해원리에 기초를 둔 기술한 새로운 방법은 순수 분리 균주는 물론 colony 계수에 사용되는 평판배지상에서도 세균의 체외세포 효소 특성을 정량적으로 측정 가능하게 한다. 본 새로운 방법을 이용하여 담수 생태계와 해양 퇴적토내 종속영양세균의 체외 효소 활성을 측정하여 고찰하였다.

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PURIFICATION AND PROPERTIES OF EXTRACELLULAR NUCLEASE(S) FROM RUMEN CONTENTS OF BUBALUS BUBALIS

  • Sinha, P.R.;Dutta, S.M.
    • Asian-Australasian Journal of Animal Sciences
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    • 제3권2호
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    • pp.115-120
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    • 1990
  • Extracellular nuclease(s) in buffalo rumen fluid were purified from strained rumen fluid by a procedure involving Seitz filtration, acetone fractionation and gel filtration on Sephadex G-100. The enzyme resolved into two peaks exhibiting both DNase and RNase activities. The molecular weight of enzyme corresponding to peaks I and II were approximately 30,000 and 12,000 respectively. The properties of enzymes from the two peaks, however, were same. Optimum temperature for both DNase and RNase activities was at $50^{\circ}C$. Whereas DNase activity was stable upto $60^{\circ}C$, RNase activity was stable only up to $50^{\circ}C$. DNase activity recorded two pH optima, one at pH 5.5 and the other at pH 7.0. RNase activity recorded a broad pH optimum between pH 6.0-8.0. pH stability of the enzyme coincided with pH optima for both the activities. DNase activity was stimulated by $Mg^{2+}$ and $Mn^{2+}$ and inhibited by $Fe^{2+}$, $Zn^{2+}$, $Hg^{2+}$ and $Ag^+$. RNase activity was also stimulated by $Mg^{2+}$ and $Mn^{2+}$ and inhibited by $Cu^{2+}$, $Fe^{2+}$, $Zn^{2+}$, $Hg^{2+}$ and $Ag^+$. Reducing agents stimulated both the activities.

Molecular Cloning, Purification, and Characterization of an Extracellular Nuclease from Aeromonas hydrophila ATCC14715

  • Nam, In-Young;Myung, Hee-Joon;Joh, Ki-Seong
    • Journal of Microbiology and Biotechnology
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    • 제14권1호
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    • pp.178-181
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    • 2004
  • A gene encoding an extracellular nuclease was cloned from Aeromonas hydrophila strain ATCC14715. The gene was overexpressed and the enzyme was purified by fusing to maltose binding protein. It was shown that the protein possessed DNase activity on both single-stranded and double-stranded DNAs. It exhibited both endo- and exonuclease activities. It was also shown that the protein had an RNase activity. Possible roles of this extracellular enzyme in the A. hydrophila life cycle are discussed.

침엽수 혼합목분에서 배양조건을 달리한 Fomitopsis palustris 의 균체외효소 활성 변화 (Effect of Various Culture Conditions on the Activity of Extracellular Enzymes from Fomitopsis palustris, Brown Rot Fungi)

  • 이영민;최두열;김현정;윤정준;김영숙
    • Journal of Forest and Environmental Science
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    • 제24권1호
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    • pp.53-59
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    • 2008
  • 목질 바이오매스(wood biomass)를 효율적으로 분해하는 최적 배양조건을 확립하기 위하여 국내 침엽수 4수종(소나무, 낙엽송, 잣나무, 리기다소나무)의 목질바이오매스를 혼합하여 목분의 적정입도, 배양기간, 기질첨가농도별 균주의 균체외효소 활성을 측정하였다. 연구 결과, 혼합목분을 기질로 했을때 Fomitopsis palustris가 분비하는 균체외효소 활성은 효소 종류에 따라 기질농도, 목분입도, 및 배양기간등의 배양조건에 따라 다른 효소활성 값을 나타냈다. 침엽수 혼합목분을 기질로 했을 때 대부분의 cellulase 분비를 유도하는 최적 조건은 목분 입도, 80~100 mesh, 기질 농도는 7.5%가 적정한 것으로 밝혀졌다. 또한 최적의 목분입도 및 투입농도 조건에서 배양기간은 4~8주로 적절한 것으로 구명되었다.

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Inactive extracellular superoxide dismutase disrupts secretion and function of active extracellular superoxide dismutase

  • Jeon, Byeong-Wook;Kim, Byung-Hak;Lee, Yun-Sang;Kim, Sung-Sub;Yoon, Jong-Bok;Kim, Tae-Yoon
    • BMB Reports
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    • 제44권1호
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    • pp.40-45
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    • 2011
  • Extracellular superoxide dismutase (EC-SOD) is an antioxidant enzyme that protects cells and tissues from extracellular damage by eliminating superoxide anion radicals produced during metabolism. Two different forms of EC-SOD exist, and their different enzyme activities are a result of different disulfide bond patterns. Although only two folding variants have been discovered so far, five folding variants are theoretically possible. Therefore, we constructed five different mutant EC-SOD expression vectors by substituting cysteine residues with serine residues and evaluated their expression levels and enzyme activities. The mutant EC-SODs were expressed at lower levels than that of wild-type EC-SOD, and all of the mutants exhibited inhibited extracellular secretion, except for C195S ECSOD. Finally, we demonstrated that co-expression of wild-type EC-SOD and any one of the mutant EC-SODs resulted in reduced secretion of wild-type EC-SOD. We speculate that mutant EC-SOD causes malfunctions in systems such as antioxidant systems and sensitizes tissues to ROS-mediated diseases.

남극 King Geroge Island 토양의 종속영양 세균 분포상과 효소 활성도 (Distribution of Heterotrophic Bacterial Flora in Soil on the King George Island (Antarctica) and Their Enzyme Activities)

  • 김상진;이승복
    • 미생물학회지
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    • 제28권3호
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    • pp.199-203
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    • 1990
  • 남극 토양생태계 세균의 분포와 세균의 생화학적 특성을 알아보기 위해 King George섬의 세종기지(한국)와 Te-niente Jubany기지(아르헨티나) 주변의 토양 17점을 남극 하계 기간(1989년 2월) 동안 채취하여 종속영양 세균의 수오 채외 효소활성도를 측정하였다. 종속영양 세균수는 시료채취지역과 배양온도가 다른 조건에서 자란 세균의 경우에도 그 평균값이 비숫한 경향을 보이고 있으나 protease는 $4^{\circ}C$에서 자란 세균의 경우 92%로서 특히 높게 나타났다. 배양온도에 따라 매우 변화가 켰으며 산술평균값은 배양온도 $37^{\circ}C$, $25^{\circ}C$, $4^{\circ}C$에서 각각 $2.5\times 10^{4}$, $2.7\times 10^{7}$, $6.9\times 10^{5}$ CFU/$cm^{3}$soil로 나타났다

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Accumulation of Selenium and Changes in the Activity of Inulinase and Catalase in the Cells of Kluyveromyces marxianus on Pulsed Electric Field Treatment

  • Pankiewicz, Urszula;Jamroz, Jerzy
    • Journal of Microbiology and Biotechnology
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    • 제20권7호
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    • pp.1101-1106
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    • 2010
  • Pulsed electric field (PEF) of 1Hz, 1.5 kV, and 1ms increased the activities of catalase and inulinase over the whole range of applied Se concentrations compared with the non-treated cultures. A significant effect of selenium concentration (in the range of 5-14 ${\mu}g/ml$) on both intra- and extracellular enzyme activities was noted. At a Se concentration of 10 ${\mu}g/ml$, the activities of intra- and extracellular inulinases and extracellular catalase in the PEF-treated cultures reached the maximum of 71 U/g d.m., 46 U/g d.m., and approx. 8 U/ml, respectively. The maximum activity of intracellular catalase of approx. 6 U/ml (with and without PEF) was recorded at 5 ${\mu}g$ Se/ml. Further increasing of selenium concentration caused a decrease in the activity of the enzymes.

Differences in Biogeochemical Properties and Microbial Activities in Stream Segments with Changes in Land-use Type

  • Kim, Jinhyun;Jang, Inyoung;Lee, Hyunjin;Kang, Hojeong
    • Ecology and Resilient Infrastructure
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    • 제2권3호
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    • pp.247-254
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    • 2015
  • Changes in land-use type can affect soil and water properties in stream ecosystems. This study examined the effects of different land-use types on biogeochemical properties and microbial activities of a stream. We collected water and sediment samples in a stream at three different sites surrounded by varying land-use types; a forest, a radish field and a rice paddy. Nitrogen contents, such as nitrate, nitrite and total nitrogen in the stream water body, showed significant differences among the sampling sites. The highest nitrogen values were recorded at the site surrounded by cropland, as fertilizer runoff impacted the stream. Soil organic matter content in the sediment showed significant differences among sites, with the highest content exhibited at the forest mouth site. These differences might be due to the organic matter in surrounding terrestrial ecosystems. Microbial activities determined by extracellular enzyme activities showed similar values throughout all sites in the water body; however, the activities in the sediments exhibited the highest values near the forest site and mirrored the soil organic matter content values. From these results, we conclude that different land-use types are important factors affecting water and sediment properties in stream ecosystems.

고효율 효소를 분비하는 균주의 선발 및 신문고지의 효소탈묵 특성(제4보) -고지탈묵용 Fungal Cellulase와 Xylanase의 생산- (Screening of Microorganisms Secreted High Efficient Enzymes and Properties of Enzymatic Deinking for Old Newsprint(IV) -Culture conditions of fungal extracellular enzyme production for biological deinking system-)

  • 박성철;강진하;이양수
    • 펄프종이기술
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    • 제37권1호
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    • pp.38-46
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    • 2005
  • This study was focused on the optimum culture condition in CMCase, FPase and xylanase activities of two fungal strains that secret extracellular enzymes for using enzymatic deinking agent to old newsprint. The results of this study were as follows. When Fusarium pallidoroseum was grown on the medium, containing of rice bran+xylan $2.0\%,\;peptone\;0.6\%,\;KH_2PO_4\;0.075\%\;and\;MnSO_4\;0.06\%\;with\;pH\;9.0,\;at\;29^{\circ}C$ for 6 days, the quantitative degree of extracellular enzyme production was the highest. Optimum culture condition for Aspergillus niger was pH 5.0, $27^{\circ}C$ incubating temperature and 7 days incubation period on liquid medium, containing of CMC+xylan $2.5\%,\;yeast\;extract\;0.4\%,\;K_3PO_4\;0.05\%\;and\;CaCl_2+FeSO_4\;0.08\%$. Aspergillus niger was fairly higher FPase and xylanase activities than Trichoderma reesei ATCC 28217.