• Title/Summary/Keyword: 토양 효소 활성

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Ketoprofen ethyl ester에 대해 높은 광학 선택성을 갖는 (R)- 과 (S)-stereospecfic esterase들의 클로닝과 서열분석 및 발현

  • Kim, Ji-Yeon;Choe, Gi-Seop;Kim, Geun-Jung;Yu, Yeon-U
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.625-628
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    • 2001
  • To isolate novel strains that hydrolyzed the rac-ketoprofen ethyl ester to ketoprofen in the stereospecific manner, we screened broad ecological niches and soil samples in which the activity was expected to be found. From thousands of strains, we isolated a Pseudomonas sp. S34 producing a (S)-stereospecific esterase, and a thermostable esterase with (R)-form selectivity was also 。 btained from Bacillus stearothermophilus JYl44. To further analyse the gene structure and to induce a high level expression, two genes from each strain were cloned and sequenced. BLAST search results with the esterase gene from 534 revealed that both of gene (80-84 %) and amino acid sequences (89- 95 %) were highly conserved in the related esterases from Pseudomonas strains (fluorescens and aeruginosa). The thermostable esterase from JY144, however, revealed a relative low homology (45-52 %) to other esterase and/or lipase from related strains. Obviously, a complete conversion with pure enantiomer (R - or S) were readily achieved by recombinant clones expressing either (R)- or (S)- stereospecific esterase.

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Characteristics of $\beta$-Galactosidase with High Transgalactosylation Activity Produced by Penicillium sp. KFCC 10888. (Penicillium sp. KFCC 10888이 생산하는 갈락토스 전이활성이 우수한 $\beta$-Galactosidase의 특성)

  • 인만진;채희정
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.40-44
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    • 1998
  • A Penicillium strain which produces $eta$-galactosidase with high transgalactosylation activity, was isolated from soil and registered as Penicillium sp, KFCC 10888. When $eta$-galactosidase from Penicillium sp. KFCC 10855 reacted with 40% lactose, transgalactosylation ratio reached up to 70% at the 73% conversion of initial lactose. The biosynthesis of the enzyme in Penicillium sp. KFCC 10888 was not induced by lactose. The soybean meal was an effective component of the culture medium. The optimum pH and temperature for transgalactosylation were 4.0 and 55$^{\circ}C$, respectively. The production of galactooligosaccharides was in proportion to the initial lactose concentration. When the enzyme reacted with 40% lactose (pH 4.0) at 55$^{\circ}C$, the concentration of galactooligosaccharides increased up to 40% of total solid concentration.

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Soil Microbial Community Analysis using Soil Enzyme Activities in Red Pepper Field Treated Microbial Agents (토양효소활성을 이용한 미생물제제 처리 고추경작지의 토양미생물군집 분석)

  • Kim, Yo-Hwan;Lim, Jong-Hui;An, Chang-Hwan;Jung, Byung-Kwon;Kim, Sang-Dal
    • Journal of Applied Biological Chemistry
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    • v.55 no.1
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    • pp.47-53
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    • 2012
  • Increasing concerns over green farming technology, plant growth promoting rhizobacterium (PGRP) having growth promoting as well as plant disease suppressing properties was recently preferred to use for biological control of plant pathogens infecting plant. We measured the influence of the selected microbial consortium agents-a mixture of PGPR strains-, commercial bio-fungicide, and chemical pesticides on soil microbial community in red pepper field. The activities of soil enzyme such as dehydrogenase, urease, phosphatase, ${\beta}$-glucosidase, and cellulase were analyzed to investigate that of soil microbial community. We also measured plant length, main stem, stem diameter, number of branches and yields of red-pepper in order to observe the red pepper growth promotion. The results of measuring enzyme activities were dehydrogenase 3.5584 ${\mu}g$ TPF $g^{-1}h^{-1}$, urease 15.8689 ${\mu}g$ $NH_4{^-}N$ $g^{-1}h^{-1}$, phosphatase 0.5692 ${\mu}g$ PNP $g^{-1}h^{-1}$, ${\beta}$-glucosidase 2.4785 ${\mu}g$ PNP $g^{-1}h^{-1}$, and cellulase 86.1597 ${\mu}g$ glucose $g^{-1}h^{-1}$ in the soil treated with the microbial consortium agents, so it came out to be very active in the soil. Observing the growth of red-peppers, the main-stem length and the stem diameter were 6.1% and 8.1% higher in the soil treated with the selected microbial consortium agent than the chemical pesticides. After harvesting, yields were 7.3% higher in the soil treated with selected microbial consortium agents than the chemical pesticides. These results showed that microbial consortium agents contribute to increasing soil microbial diversity, growth promoting, and yield of red pepper.

호산성 방선균과 내중성 방선균

  • 강희일
    • The Microorganisms and Industry
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    • v.15 no.2
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    • pp.2-7
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    • 1989
  • Waksman과 Heinrich(1943)에 의해 Streptomyces속이 기술되고 actinomycin, streptomycin등의 많은 항생물질이 발견되면서 방선균(actinomycetes)은 산업적으로 중요한 미생물이 되어 왔으며, 근래에는 항생물질 뿐만 아니라 효소, 효소저해제, 면역조절물질 등 소위 생리활성물질(bioactive compound)의 생산균으로서 주목을 받고 있다. 그러나 지금까지 대부분의 스크리닝이나 연구의 대상은 주로 중성 pH에서 잘 자라는 호중성 방선균(neutrophilic actionmycetes)이었지만 산성이나 알칼리성 토양, 고온, 고열, 고압 등과 같은 특수환경에서 분리된 균주나 희귀 방선균(rare actinomyces)에 대한 관심도 점점 높아지고 있다. 산성 토양에서 분리된 방선균에 관한 연구는 Jensen(1928)의 연구로부터 시작하였지만 Lonsdale(1985)이 침엽수림, 산성탄광 등에서 분리된 방선균을 수리분류할 때까지 활발하게 이루어지지 못하였다. 본 고에서는 호산성 방선균위 분포, 생리적 특성, 수리분류학적 연구, 화학적 분류(chemotaxonomy), 산업적 응용가능성 등을 살펴보고자 한다.

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Domain Function and Relevant Enzyme Activity of Cycloinulooligosaccharide Fructanotransferase from Paenibacillus polymyxa (Paenibacillus polymyxa Cycloinulooligosaccharide Fructanotransferase의 효소 활성에 미치는 각 Domain의 역할)

  • You Dong-Ju;Park Jung-Ha;You Kyung-Ok;Nam Soo-Wan;Kim Kwang-Hyeon;Kim Byung-Woo;Kwon Hyun-Ju
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.278-287
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    • 2006
  • Cycloinulooligosaccharide fructanotransferase (CFTase) converts inulin into cycloinulooligosaccharides (cyclofructan, CF) of ${\beta}-(2{\to}1)$-linked D-fructofuranose as well as hydrolysis of cyclofructan. Sequences analysis indicated that CFTase was divided into five distinct regions containing three repeated sequences (R1, R3, and R4) at the N-terminus and C-terminus. Each domain function was investigated by comparison of wild type CFTase enzyme (CFT148) and deletion mutant proteins (CFT108: R1 and R3 deletion; CFT130: R4 deletion; and CFT88: R1, R3, and R4 deletion) of CFTase. The CFT108 mutant had both CFTase and CF hydrolyzing activity as CFT148 did. CFTase activities and CF hydrolysing activities were disappeared in CFT130 and CFT88 mutants. These results indicated that the C-terminal R4 region of P. polymyxa CFTase is necessary for cyclization and hydrolyzing activity.

Characterization of Alternaria alternata ${\alpha}-Amylase$ (Alternaria alternata ${\alpha}-Amylase$의 특성에 관한 연구)

  • Chung, Sang-Jin;Hwang, Baik
    • The Korean Journal of Mycology
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    • v.24 no.1 s.76
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    • pp.8-16
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    • 1996
  • The ${\alpha}-amylase$ of Alternaria alternata was purified through ammonium sulfate precipitation, dialysis and Sephadex G-100 column chromatography. One single band was obtained in SDS-polyacrylamide gel electrophoresis. The optimum pH for enzyme activity was 5.0 and the enzyme activity was maintained at $3.6{\sim}7.0$pH range. The optimum temperature for ${\alpha}-amylase$ activity was $40^{\circ}C$ and 71% of the activity was still maintained until 30 min after heating at $80^{\circ}C$. The ${\alpha}-amylase$ was slightly activated by $Mn^{2+},\;Zn^{2+}\;and\;Sn^{2+}$, but inhibited by $Ba^{2+},\;Pb^{2+},\;Co^{2+}\;and\;Ag^{1+}$. The $Hg^{2+}\;and\;Ag^{2+}$ slightly inhibited the activity of the enzyme at concentrations of $10^{-3}\;and\;10^{-4}M$. The Michaelis constant $(K_m)$ to soluble starch was $6.50{\times}10^{-2}M$ and inhibition constant $(K_i)$ by the 1mM EDTA was $8.0{\times}10^{-2}M$. The inhibition of this enzyme by EDTA was competitive one.

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Pseudomonas species에 의한 inulase 생산 기초연구

  • 이태경;김형수;양한철
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.527.2-527
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    • 1986
  • 토양시료로부터 높은 활성의 extracellular inulase를 생산하는 균주를 분리하여 분리균의 형태적 내지는 생리적 성질을 조사한 결과 Pseudomonas species로 동정되었다. 또한 Pseudomonas species로 동정된 본 분리균은 initial pH가 7.0인 1% inulin, 1.5% cornsteepliquor 0.8% (NH$_4$)$_2$ HPO$_4$ 및 0.003% FeSO$_4$ 7$H_2O$의 조성을 지닌 배지, 배양온도 45$^{\circ}C$ 및 배양시간 72시간 등의 배양조건에서 효소활성이 배양액 $m\ell$ 당 6.1 units로서 가장 높은 효소생산량을 나타내었다.

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Pantoea spp.에서 분리한 호냉성 ${\beta}-Galactosidase$의 생화학적 특성 및 우유 내 유당분해 활성

  • Choe, Jae-Won;Lee, Seung-Bae;Choe, Seok-Ho
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.10a
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    • pp.384-387
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    • 2004
  • 겨울철 토양에서 ${\beta}-Galactosidase$를 생산하는 균주를 분리하였으며 동정한 결과 그람 음성 간균이고 Pantoea spp. 로 확인되었다. Pantoea spp. 균주의 세포 추출물로부터 DEAE-Sephacel chromatography와 affinity chromatography를 이용하여 ${\beta}-Galactosidase$를 분리하였다. Pantoea spp. 의 ${\beta}-Galactosidase$의 반응 최적 온도는 $45^{\circ}C$이이고 최적 pH는 $5.5{\sim}7.5$이고 열안정성을 조사한 결과 $45^{\circ}C$이상의 온도에서 불활성 되는 것으로 나타났고 E. coli에서 분리된 효소보다 저온에서의 활력이 좋았지만 상업적인 효소인 Kluyveromyces lactis (Validase) 보다는 낮았다.

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Cellulose Degradation and Extracellulat Enzymatic Activity of the Mud Flat in Sunchon Bay (순천만 갯벌 토양의 섬유소 분해능 및 체외효소 활성)

  • 백근식;최지혁;성치남
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.130-135
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    • 2000
  • Decomposition rate of organic matiter in the mud flat of Sunchon Bay was estimated. Physicochemical parameters, cellulose degradation rate. distribution of heterotrophic bacteria, and extracellular enzymatic activities were measured from August 1997 to July 1998. Soil temperatures, water contents, concentration of $PO_4$-P and organic matter were -1-~$30^{\circ}C$, 42.1-53.1%, 0.0779-0.1961 mgig and 1.99-7.64%, respectively. Decomposition rate of cellulose film ranged from 7.7 to 100%imonth, high in summer and low in winter. The number of heterotrophic bacteria ranged from $0.87{\times}10^6 to 3.6{\times}10^7 $CUFsIg dq soil. Enzymatic activities of phosphatase, $\alpha$-D-gluEosidase, $\beta$-D-glucosidase and cellobiohydrolase, which were measured as decomposition rate of methylumbelliferyl(MLiF)-substrate, were 152.23-1779.80 nMIhr, 2.67-202.18 nM/hr, 5.03-258.26 M h r and 3.42-63.07 nM/hr, respectively Cellulose degradaaon rate and extracellular extracellular enzymatic activities were conelated with each other, and showed high correlation coefticiency with soil temperature.

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Studies on the Isolation and Identification of Xylanase and Mannanase Producing Aspergillus niger (Xylanase와 Mannanase를 생산하는 Aspergillus niger의 분리와 동정에 관한 연구)

  • Kim, Byoung-Suk;Cho, Jin-Kook;Song, Jin-Ook;Lee, Hak-Kyo;Hwang, Seong-Gu
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.427-432
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    • 2009
  • This study was undertaken to screen a high xylanase and mannanase producing microbes. In the first experiment, screening was undertaken against 50 samples of microorganisms having xylanase and mannanase activities from soil and fallen leaves. The screening process has focused on picking out fungi having high xylanase and mannanase activities under the solid-state fermentation. The xylanase and mannanase activities of 6 screened microbes were 0.9~1.6 unit/mL and 0.2~0.4 unit/mL, respectively, under the submerged fermentation condition. However, under the solid-state fermentation, xylanase and mannanase activities were 103.7~220.0 unit/g and 20.1~40.3 unit/g, respectively. Finally one microbe (E-3) was selected and its xylanase and mannanase activities were 197.3 unit/g and 39.9 unit/g, respectively. The morphological and molecular biological classification of E-3 showed 99% homology with the Aspergillus niger.