• 제목/요약/키워드: hydrolytic activity

검색결과 193건 처리시간 0.025초

토양에서 분리한 Aeromonas sp 로 부터 \beta-mannanase 유전자의 클로닝 (Cloning of \beta-mananase gene from Aeromonas sp. in E. coli)

  • 박봉환;강대경;김하근
    • 한국미생물·생명공학회지
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    • 제29권4호
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    • pp.201-205
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    • 2001
  • 자연계로부터 locust bean gum이 들어 있는 선택배지에서 투명환을 형성하는 mannan 분해능을 갖는 미생물을 스크링 하였고 이를 Aeromonas sp. 로 동정하였다. Aero monas sp. 로부터 염색체 DNA를 분리하여 EcoRI으로 절단하여 대장균에 형질전환한 후 대장균 콜로니 주위에서 locust bean gum을 분해하여 투명환을 형성하는 대장균을 찾아냈다. 형질전환된 대장균으로부터 플라스미드를 분리하여 동일한 제한효소를 처리하여 확인한 결과 10 kd의 Aeromonas sp. 염섹체 DNA에 $\beta$-mannanase 유전자 존재하였다. 대장균에서 발현된 $\beta$-mannanase 의최적 반응 pH와 최적 반응온도는 각각 6.0과 $50^{\circ}C$로서 모균인 Aeromonas sp. 와 동일하였다.

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5-Fluorouracil 유도체 합성 및 항암작용 (Synthesis and Antitumor Activity of $N^1$-derivatives of 5-Fluorouracil)

  • 이희주;신혜순;진현숙;김지현;김종국
    • 약학회지
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    • 제37권1호
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    • pp.89-94
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    • 1993
  • In order to find out a proper connecting bridge between 5-fluorouracil(5-FU) and a macromolecule such as a polypeptide, potentially hydrolytic N$^{1}$-derivartives of 5-FU have been systhesized and evaluated for their biological activity. When tested with in vitro leukemic L$_{1210}$ cells all the obtained derivartives exhibited slightly higher antitumor activity than the parent 5FU. Among them the N$^{1}$ -carbamoyl analogue 2 and N$^{1}$-acetamido analogue 6b showed 50% inhibition of the L$_{1210}$ cell growth at the concentrations of 5.01$\times$10$^{-8}$M and 1.03$\times$10$^{-7}$M, respectively. When tested against sarcoma 180 tumor cells inoculated into mice, the compounds 2 and 6b exhibited, respectively, 62% and 54% inhibition of the solid tumor growth at the 5-time doses of 100 mg/kg/day. Both compounds, N$^{1}$-carbamoyl analogue 2 and N$^{1}$-acetamido analogue 6b, realeased the parent 5-FU when incubated in the L$_{1210}$ cell cultural media for 5 hrs.

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토양효소활성 측정법을 이용한 화약류 오염토양 독성평가 (The Toxicity Assessment of Explosives Contaminated Soil using Soil Microbial Activity Tests)

  • 김문경;정재웅;남경필
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제20권6호
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    • pp.37-45
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    • 2015
  • This study was conducted to determine the toxic effect of TNT and RDX on indigenous soil microbes by measuring enzymatic activity. Denitrification activity, dehydrogenase activity, phosphatase activity, and fluorescein diacetate hydrolytic activity were determined for military firing range, field, and paddy soils exposed to TNT, and RDX from 0 to 1,000 mg/kg and 0 to 4,000 mg/kg, respectively, for 2, 4, and 8 weeks. Soil microbial enzymatic activities decreased with higher TNT and RDX concentration and longer exposure time. Microbial enzymatic activities of firing range soil were higher than field and paddy soils, indicating that indigenous microbes in firing range might have been adapted to TNT and RDX due to pre-exposure of the explosives. In addition, the toxicity of TNT and RDX decreased with higher organic matter because TNT and RDX tend to absorb to soil organic matter. No Observable Effect Concentration (NOEC) values of each microbial enzymatic activity were derived by the geometric mean of NOECs from exposure times (2, 4, and 8 weeks) and soil types (firing range, field, paddy soil). The derived NOECs ranged from 45.3 to 55.2 mg/kg for TNT and 286 to 309 mg/kg for RDX.

Analysis of the Involvement of Chitin-Binding Domain of ChiCW in Antifungal Activity, and Engineering a Novel Chimeric Chitinase with High Enzyme and Antifungal Activities

  • Huang, Chien-Jui;Guo, Shu-Huei;Chung, Shu-Chun;Lin, Yu-Ju;Chen, Chao-Ying
    • Journal of Microbiology and Biotechnology
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    • 제19권10호
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    • pp.1169-1175
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    • 2009
  • An antifungal chitinase, ChiCW, produced by Bacillus cereus 28-9 is effective against conidial germination of Botrytis elliptica, the causal agent of lily leaf blight. ChiCW as a modular enzyme consists of a signal peptide, a catalytic domain, a fibronectin type-III-like domain, and a chitin-binding domain. When two C-terminal domains of ChiCW were truncated, $ChiCW{\Delta}FC$ (lacking the chitin-binding domain and fibronectin type III-like domain) lost its antifungal activity. Since $ChiCW{\Delta}C$ (lacking the chitin-binding domain) could not be expressed in Escherichia coli as $ChiCW{\Delta}FC$ did, a different strategy based on protein engineering technology was designed to investigate the involvement of the chitin-binding domain of ChiCW ($ChBD_{ChiCW}$) in antifungal activity in this study. Because ChiA1 of Bacillus circulans WL-12 is a modular enzyme with a higher hydrolytic activity than ChiCW but not inhibitory to conidial germination of Bo. elliptica and the similar domain composition of ChiA1 and ChiCW, the C-terminal truncated derivatives of ChiA1 were generated and used to construct chimeric chitinases with $ChBD_{ChiCW}$. When the chitin-binding domain of ChiA1 was replaced with $ChBD_{ChiCW}$, the chimeric chitinase named ChiAAAW exhibited both high enzyme activity and antifungal activity. The results indicate that $ChBD_{ChiCW}$ may play an important role in the antifungal activity of ChiCW.

염화갈륨, 인듐 및 탈륨 함유 이분자형 키랄 살렌 촉매에 의한 라세믹 알킬 글리시딜레이트 유도체의 비대칭 가수분해반응 (Hydrolytic Kinetic Resolution of Racemic Alkyl-glycidyl Derivatives by using Dimeric Chiral Salen Catalyst Containing Ga, In and TlCl3)

  • 신창교;카테카 라울;김건중
    • 공업화학
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    • 제18권3호
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    • pp.218-226
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    • 2007
  • 키랄성 말단기의 에폭사이드는 키랄중간체나 여러 출발물질로서 다양하게 이용되기 때문에 입체선택적인 합성방법은 학술적으로나 산업적인 관점에서 대단히 흥미롭다. 본 연구에서는 염화갈륨, 인듐 및 탈륨을 함유한 2분자형의 키랄성 코발트 살렌 촉매를 새로이 합성하고 그 특성을 평가하였다. 촉매가 2분자형을 갖는지의 직접적인 증거를 질량분석과 EXAF분석을 통하여 제시하였다. 합성한 촉매는 여러 종류의 에테르 및 에스테르 그룹을 함유한 에폭사이드 유도체의 가수분해의 속도차에 의한 비대칭 고리열림반응에 적용하여 그 활성과 선택성을 조사하였다. 합성이 용이한 2분자구조의 살렌착체 촉매는 물을 친핵체로 하는 라세믹 에폭사이드의 고리 열림을 통하여 99%ee 이상을 나타낼 정도의 매우 높은 광학선택성을 보였다. 2분자형의 살렌 촉매는 단분자형의 촉매에 비하여 촉매량을 적게 첨가하여도 현저히 향상된 촉매활성을 나타내었다. 본 연구에서 적용한 촉매시스템은 키랄 에폭사이드 및 1,2-디올 중간체의 제조에 매우 효과적이었다

Biological Control of Bacterial Fruit Blotch of Watermelon Pathogen (Acidovorax citrulli) with Rhizosphere Associated Bacteria

  • Adhikari, Mahesh;Yadav, Dil Raj;Kim, Sang Woo;Um, Young Hyun;Kim, Hyun Seung;Lee, Seong Chan;Song, Jeong Young;Kim, Hong Gi;Lee, Youn Su
    • The Plant Pathology Journal
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    • 제33권2호
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    • pp.170-183
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    • 2017
  • Bacterial fruit blotch (BFB), which is caused by Acidovorax citrulli, is a serious threat to watermelon growers around the world. The present study was conducted to screen effective rhizobacterial isolates against 35 different A. citrulli isolates and determine their efficacy on BFB and growth parameters of watermelon. Two rhizobacterial isolates viz. Paenibacillus polymyxa (SN-22), Sinomonas atrocyanea (NSB-27) showed high inhibitory activity in the preliminary screening and were further evaluated for their effect on BFB and growth parameters of three different watermelon varieties under greenhouse conditions. The greenhouse experiment result revealed that SN-22 and NSB-27 significantly reduced BFB and had significant stimulatory effect on total chlorophyll content, plant height, total fresh weight and total dry weight compared to uninoculated plants across the tested three watermelon varieties. Analysis of the 16S ribosomal RNA (rRNA) sequences revealed that strains SN-22 belong to P. polymyxa and NSB-27 to S. atrocyanea with the bootstrap value of 99% and 98%, respectively. The isolates SN-22 and NSB-27 were tested for antagonistic and PGP traits. The result showed that the tested isolates produced siderophore, hydrolytic enzymes (protease and cellulose), chitinase, starch hydrolytic enzymes and they showed phosphate as well as zinc solubilizing capacity. This is the first report of P. polymyxa (SN-22) and S. atrocyanea (NSB-27) as biocontrol-plant growth promoting rhizobacteria on watermelon.

Characterization of the pcbD Gene Encoding 2-Hydroxy-6-Ox0-6-Phenylgexa-2,4-Dienoate Hydrolase from Pseudomonas sp. P20

  • Lim, Jong-Chul;Lee, Jeong-Rai;Lim, Jai-Yun;Min, Kyung-Rak;Kim, Chi-Kyung;Ki, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • 제10권2호
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    • pp.258-263
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    • 2000
  • 2-Hydroxy-6-oxo-6phenylhexa-2,4-dienoate (HOPDA) hydrolase catalyzes the hydrolytic cleavage of HOPDA to bemzpate and 2-hydroxypenta-2, 4-dienoate (HPD) during microbial catabolism of biphenyl and polychlorinated biphenyls. A HOPDA hydrolase gene (pcbD) was isolated from the genomic library of Pseudomonas sp. P20 and designated as pCNUO1201; a 7.5-kb XbaI DNA fragment from Pseudomonas sp. P20 was inserted into the pBluescript SK(+) XbaI site. E. coli HB101 harboring pCNU1201 exhibited HOPDA hydrolase activity. The open reading frame (ORF) corresponding to the pcbD gene consisted of 855 base pairs with an ATG initiation codon and a TGA termination codon. The ORF was preceded by a rebosome-binding sequence of 5'-TGGAGC-3' and its G+C content was 55 mol%. The pcbD gene of Pseudomonas sp. P20 was located immedeately downstream of the pcbC gene encoding 2,3- dihydroxybiphenyl 1,2-dioxygenase, and approximately 4-kb upstream of the pcbE gene encoding HPD hydratase. The pcbK gene was able to encode a polypeptide with a molecular weight of 31,732 containing 284 amino acid residues. The deduced amino acid sequence of the HOPDA hydrolase of Pseudomonas sp. P20 exhibited high identity (62%) with those of the HOPDA hydrolases of P. putida KF715, P. pseudoalcaligenes KF707, and Burkholderia cepacia LB400, and also significant homology with those of other hydrolytic enzymes including esterase, transferase, and peptidase.

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BF3 함유 이분자형 키랄 살렌 촉매에 의한 고광학순도의 에피클로로히드린 합성 (Synthesis of Optically pure Epichlorohydrine using Dimeric Chiral Salen Catalyst Containing BF3)

  • 이광연;카테카라울;김건중
    • 공업화학
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    • 제18권4호
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    • pp.330-336
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    • 2007
  • 본 연구에서는 $BF_3$를 함유한 키랄성 코발트 살렌 촉매를 합성하고 비대칭촉매특성을 평가하였다. 촉매의 구조를 결정하기 위하여 NMR, UV 및 ESCA분석을 수행하였다. 합성한 촉매는 여러 종류의 에폭사이드 유도체의 가수분해 속도차에 의한 비대칭 고리열림반응에 적용하여 그 활성과 선택성을 조사하였다. 살렌과 $BF_3$의 비를 다르게 하여 합성한 촉매는 전혀 다른 촉매 활성을 나타내었으며, 그 비를 2 : 1로 하여 합성한 2분자구조의 살렌착체 촉매는 물을 친핵체로 하는 라세믹 에폭사이드의 고리 열림을 통하여 99 %ee 이상의 매우 높은 광학선택성을 보였다. 특히 2분자형의 살렌 촉매는 단분자형의 촉매에 비하여 촉매량을 적게 첨가하여도 현저히 향상된 촉매활성을 나타내었고 얻어진 생성물의 분리과정에서 라세믹화를 유발시키지 않았다. 본 연구에서 적용한 촉매시스템은 키랄 에폭사이드 및 1,2-디올 중간체의 제조에 매우 효과적이었다.

Serratia marcescens S3-R1이 생산한 효소에 의한 유청단백질 가수분해물의 특성과 면역조절 활성 (Whey protein hydrolytic properties and its immunomodulation activity by produced enzyme from Serratia marcescens S3-R1)

  • 유재민;렌친핸드;정석근;배형철;남명수
    • 농업과학연구
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    • 제40권3호
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    • pp.221-226
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    • 2013
  • Degrees of hydrolysis by alkaline protease produced from Serratia marcescens S3-R1 is 3.95-6.30% of whey proteins during 5, 15, 30, 60, 90, 120,180, 240 min incubation at $40^{\circ}C$. Proteolytic pattern of the whey proteins showed that various low molecular weight peptides were generated during the incubation periods. The biological function of in Raw 264.7 cells treated with whey protein hydrolytic peptides, anti-inflammatory effect showed exhibit in the expression of pro-inflammatory cytokines such as TNF-${\alpha}$, IL-6, COX-2 and iNOS by PCR analysis. COX-2 and iNOS gene expression inhibited in Raw 264.7 cells on whey protein hydrolysates below 3,000 dalton. The protease from Serratia marcescens S3-R1 showed a potential in production of low molecular weight whey protein hydrolysates which could be used for industrial application.

전이금속염 함유 키랄 살렌 촉매에 의한 광학선택적 에폭사이드의 합성 (Enantioselective Epoxide Synthesis on the Chiral Salen Catalyst having a Transitional Metal Salt)

  • 곽소봉;키테라 라올;김건중
    • Korean Chemical Engineering Research
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    • 제46권4호
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    • pp.769-776
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    • 2008
  • 키랄성 말단기의 에폭사이드는 키랄중간체나 여러 출발물질로서 다양하게 이용되기 때문에 그 선택적인 합성법은 매우 유용하다. 본 연구에서는 염화코발트(II), 염화철(III) 및 질산아연(II)을 각각 함유한 키랄 코발트 살렌 촉매를 새로이 합성하고 그 특성을 평가하였다. 질량분석과 EXAF분석을 통하여 형성된 촉매 착체의 구조를 평가하였다. 합성한 촉매는 방향족 에폭사이드인 스타이렌 옥사이드와 페닐글리시딜 에테르의 속도차에 의한 비대칭 가수분해적 고리열림반응과 글리시틸부틸레이트의 합성반응에 적용하여 그 활성과 선택성을 조사하였다. 합성이 용이한 전이금속염함유 살렌착체 촉매는 물을 친핵체로 하는 라세믹 에폭사이드의 고리 열림을 통하여 99%ee 이상을 나타낼 정도의 매우 높은 광학선택성을 보였으며, 적은 양의 첨가로도 높은 활성을 보였다. 본 연구에서 적용한 촉매씨스템은 키랄 에폭사이드 및 1,2-디올 중간체의 제조에 매우 효과적이었다.