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Purification and Characterization of Two Alkaline Proteases Produced by Pseudomonas sp. BK7

  • 이은구;박은희;현형환
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.667-667
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    • 2000
  • Pseudomonas sp. BK7, an alkalophile, displayed the highest growth and protease activity when grown in a fermenter which was controlled at a pH level of 9.0, and the enzyme production was significantly enganced by the increase of agitation speed. Two formas of alkaline proteases (BK7-1 and BK7-2) were fractionated and purified to near homogeneity. Protease BK7-1 was purified through CM-Sepharose CL-6B and Sephadex G-75 column chromatographies, and Protease BK7-2 was purified through CM-Sepharose CL-6B and Sephadex G-75 column chromatographies, and Protease BK7-2 was purified through CM-Sepharose CL-6B, DEAE-Sepharose, and Sephadex G-75 column chromatographies. The molecular weights of proteases BK7-1 and BK7-2 determined by gel filtration chromatography were 20,700 and 40,800, respectively. The $K_m$ value, isoelectric point, and optimum pH of protease BK7-1 were 2.55 mg/ml, 11.0 and 11.0, respectively, whereas those of protease BK7-2 were 1.57 mg/ml, 7.2, and 10.0, respectively. Both protease were practically stable in the pH range of 5-11. The optimum temperatures for the activities of both protease BK7-1 and BK7-2 were 50℃ and 45℃, respectively. About 56% of the original protease BK7-2 activity remained after being treated at 50℃ for 30 min but protease BK7-1 was rapidly inactivated at above 25℃. Both proteases were completely inhibited by phenylmethane sulfonyl fluoride, a serine protease inhibitor. Protease BK7-2 was stable against EDTA, EGTA, STP, and detergents such as SDS and LAS, whereas protease BK7-1 was found to be unstable.

Isolation and Characterization of Soil Strains Producing Glutaryl-7-Aminocephalosporanic Acid Acylase

  • Knang, Yong-Ho;Yoo, Ryong-Hoon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.2 no.2
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    • pp.105-108
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    • 1997
  • A search was undertaken to screen microorganisms that produce an enzyme capable of deacylating glutary1-7-amincephalosporanic acid to 7-aminocephalosporanic acid in soil samples. The screening was carried out by preparing enrichment cultures containing glutary-7ACA and cephalosporin C as selective carbon sources. A non-${\beta}$-lactam model compound,, glutary-p-nitroanilide, was synthesized as a substrate suitable for the rapid screening of microorganisms isolated from the enrichment cultures. Two isolates exhibiting acylase activity, designated BY7.4 and BY8.1, were identified as strains of Pseudomonas species. Pseudomonas BY8.1 showed higher acylase activity toward G1-7ACA than Pseudomonas BY7.4. Environmental conditions for the optimal acylase activity of Pseudomonas BY8.1 were shown to be pH9 and 30$^{\circ}C$.

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Fabrication of the in-plane Aligned a-Axis Oriented $YBa_2Cu_3O_{7-x}$ Thin Films (평면배향된 a-축 수직 $YBa_2Cu_3O_{7-x}$ 고온초전도 박막의 제작)

  • 성건용;서정대
    • Journal of the Korean Ceramic Society
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    • v.33 no.3
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    • pp.313-320
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    • 1996
  • We have fabricated an in-plane aligned a-axis oriented YBa2Cu3O7-x (a-YBCO) thin film on a LaSrGaO4(100) substrate with a PrBa2Cu3O7-x(PBCO) template layer by two step plused laser deposition using 308 nm XeCl excimer laser. A YBCO layer and PBCO layer grown at low temperatures were used as template layers. We have investigated the effect of the deposition temperature of template layers on the superconducting and struc-tural properties of in-plane aligned a-YBCO thin films. An optimal deposition temperature of the PBCO template layers was 630. In-plane aligned a-YBCO thin films showed an anisotropy ratio in resistivity of 11.5 and a zero resistance temperature of 88 K.

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Synthesis of Theophylline Derivatives (Theophylline 유도체의 합성)

  • Suh, Jung-Jin;Lee, Bong-Yong
    • YAKHAK HOEJI
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    • v.33 no.1
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    • pp.10-14
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    • 1989
  • As potential xanthine bronchodilators related to Doxofylline (1b), 2-(7-theophyllineethyl)-1,3-dioxolane (3), 7-(3,3-dimethoxypropyl) theophylline(5), 2-methyl, 2-(7-theophyllinemethyl)-1,3- dioxolane (8a), 2-phenyl, 2-(7-theophyllinemethyl)-1,3-dioxolane (8b) and 2-(7-theophyllinemethyl)-1,3-benzodioxolane(14) were synthesized from theophylline (1a).

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A Case of Partial Inverted Duplication of Chromosome 7q (염색체 7번 장완의 부분적 역위 중복 1례)

  • Kim, Hyun Ji;Lee, So Young;Kim, Mi Young;Baek, Chung Sun;Kim, Won Duck;Lee, Dong Seok;Kim, Doo Kwun;Choi, Sung Min
    • Clinical and Experimental Pediatrics
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    • v.46 no.5
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    • pp.510-513
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    • 2003
  • Duplication of chromosome 7q has been reported as either partial or complete. Partial 7q duplication was first described by Carpentier in 1972. Pure partial duplication of the long arm of chromosome 7 is extremely rare and only 16 cases with a pure partial duplication of different 7q segment have been described in the literature. Pure partial duplication of the long arm of chromosome 7 is characterized by growth and developmental retardation, muscular hypotonia, distinct craniofacial dysmorphic features, a short neck and skeletal abnormalities. A 3 month-old male was referred to our department of Pediatrics because of dyspnea, hypotonia and delayed development. He shows growth and developmental delay, hypertelorism, a depressed nasal bridge, low set ears, a short neck and muscular hypotonia. Karyotype revealed 46, XY, dup(7)(q36q33) by GTC-banding. We report a case of a partial inverted duplication of chromosome 7q.

Let-7c Inhibits NSCLC Cell Proliferation by Targeting HOXA1

  • Zhan, Min;Qu, Qiang;Wang, Guo;Liu, Ying-Zi;Tan, Sheng-Lan;Lou, Xiao-Ya;Yu, Jing;Zhou, Hong-Hao
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.387-392
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    • 2013
  • Objective: The aim of the present study was to explore mechanisms by which let-7c suppresses NSCLC cell proliferation. Methods: The expression level of let-7c was quantified by qRT-PCR. A549 and H1299 cells were transfected with let-7c mimics to restore the expression of let-7c. The effects of let-7c were then assessed by cell proliferation, colony formation and cell cycle assay. Mouse experiments were used to confirm the effect of let-7c on tumorigenicity in vivo. Luciferase reporter assays and Western blotting were performed to identify target genes for let-7c. Results: HOXA1 was identified as a novel target of let-7c. MTS, colony formation and flow cytometry assays demonstrated that forced expression of let-7c inhibited NSCLC cell proliferation by inducing G1 arrest in vitro, consistent with inhibitory effects induced by knockdown of HOXA1. Mouse experiments demonstrated that let-7c expression suppressed tumorigenesis. Furthermore, we found that let-7c could regulate the expression of HOXA1 downstream effectors CCND1, CDC25A and CDK2. Conclusions: Collectively, these results demonstrate let-7c inhibits NSCLC cell proliferation and tumorigenesis by partial direct targeting of the HOXA1 pathway, which suggests that restoration of let-7c expression may thus offer a potential therapeutic intervention strategy for NSCLC.

Studies on Biological Activity of Wood Extractives(VI) - Flavonoids in heartwood of Prunus sargentii - (수목추출물의 생리활성에 관한 연구(VI) - 산벚나무 심재의 Flavonoids -)

  • Lee, Hak-Ju;Lee, Sung-Suk;Choi, Don-Ha;Kato, Atsushi
    • Journal of the Korean Wood Science and Technology
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    • v.29 no.2
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    • pp.133-139
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    • 2001
  • The structures of six flavonoids isolated from heartwood of Prunus sargentii(Rosaceae) were analyzed by Mass and NMR spectrometry. These flavonoids were grouped into dihydroflavonol, flavanone, and flavanone glycoside, and identified as follows : 3,3',4',5,7-pentahydroxyflavanone(taxifolin) as a dihydroflavonol, 5-hydroxy-7-methoxyflavanone(pinostrobin), 4',5,7-trihydroxyflavanone(naringenin), 3',4',5,7-tetrahydroxyflavanone(eriodictyol), 5,7-dihydroxyflavanone(pinoccmbrin) as a flavanone and 7-hydroxyflavanone 5-O-${\beta}$-D-glucopyranoside(verecundin) as a flavanone glycoside.

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Room temperature growth of Mg on the Si(111)-7$\times$7 surface studied using STM and LEED

  • Lee, Dohyun;Kim, Sehun;Koo, Ja-Yong;Lee, Geunseop
    • Proceedings of the Korean Vacuum Society Conference
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    • 2000.02a
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    • pp.150-150
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    • 2000
  • The adsorption geometry and the electronic property of Mg grown at room temperature on the Si(111)-7$\times$7 surface with various coverages have been studied by scanning tunneling microscopy (STM) and low energy electron diffraction (LEED). At low Mg coverage, the Mg atoms preferentially adsorb at the center adatom sites of the faulted half of the Si(111)-7$\times$7 surface. The adsorbed Mg atom acts as nucleophile with respect to Si atoms thus forms a stable ionic bond with the substrate Si atoms. Above 1 Ml, the 7$\times$7 surface starts to be disrupted and an amorphous Mg overlayer is formed. The LEED shows either $\delta$7$\times$7 or 1$\times$1 pattern at this coverage. When more Mg atoms were exposed, a flat and broad {{{{ { 2} over {3 } }}}}{{{{ SQRT { 3} }}}}$\times${{{{ { 2} over {3 } }}}}{{{{ SQRT { 3} }}}}R30$^{\circ}$region evolves. A flat silicide is formed at first and multi-level Mg islands having hexagonal step edges develop with increasing coverage. The scanning tunneling spectroscopy (STS) confirms the electronic properties of these Mg films on the si(111) 7$\times$7 surface at various coverages.

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Selective Homocysteine Assay with Cucurbit[7]uril by pH Regulation

  • Bae, Won-Bin;Kim, Hee-Joon;Jhee, Kwang-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.32 no.4
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    • pp.514-521
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    • 2022
  • We report the effect of pH on the supramolecular complexation of two biothiols, viz., homocysteine (Hcy) and cysteine (Cys), with cucurbit[7]uril (CB[7]). Under basic pH conditions, Cys did not complex with CB[7], whereas Hcy efficiently complexed with CB[7], as confirmed by 1H NMR spectroscopy and Ellman's reagent (5,5'-dithio-bis(2-nitrobenzoic acid), DTNB) assay. 1H NMR and Raman spectroscopic studies revealed that, in the absence of CB[7], Hcy auto-oxidized slowly (~36 h) to homocystine (HSSH) under basic pH conditions. However, the rate of Hcy oxidation increased by up to 150 fold in the presence of CB[7], as suggested by the DTNB assay. Thus, supramolecular complexation under basic pH conditions led to the formation of a HSSH-CB[7] complex, and not Hcy-CB[7]. The results indicate that Hcy is rapidly oxidized to HSSH under the catalysis of CB[7], which acts as a reaction chamber, in basic pH conditions. Our studies suggest that Hcy concentration, a risk factor for cardiovascular disease, can be selectively and more easily quantified by supramolecular complexation with CB [7].

Rice 7-Hydroxymethyl Chlorophyll a Reductase Is Involved in the Promotion of Chlorophyll Degradation and Modulates Cell Death Signaling

  • Piao, Weilan;Han, Su-Hyun;Sakuraba, Yasuhito;Paek, Nam-Chon
    • Molecules and Cells
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    • v.40 no.10
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    • pp.773-786
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    • 2017
  • The loss of green coloration via chlorophyll (Chl) degradation typically occurs during leaf senescence. To date, many Chl catabolic enzymes have been identified and shown to interact with light harvesting complex II to form a Chl degradation complex in senescing chloroplasts; this complex might metabolically channel phototoxic Chl catabolic intermediates to prevent oxidative damage to cells. The Chl catabolic enzyme 7-hydroxymethyl Chl a reductase (HCAR) converts 7-hydroxymethyl Chl a (7-HMC a) to Chl a. The rice (Oryza sativa) genome contains a single HCAR homolog (OsHCAR), but its exact role remains unknown. Here, we show that an oshcar knockout mutant exhibits persistent green leaves during both dark-induced and natural senescence, and accumulates 7-HMC a and pheophorbide a (Pheo a) in green leaf blades. Interestingly, both rice and Arabidopsis hcar mutants exhibit severe cell death at the vegetative stage; this cell death largely occurs in a light intensity-dependent manner. In addition, 7-HMC a treatment led to the generation of singlet oxygen ($^1O_2$) in Arabidopsis and rice protoplasts in the light. Under herbicide-induced oxidative stress conditions, leaf necrosis was more severe in hcar plants than in wild type, and HCAR-overexpressing plants were more tolerant to reactive oxygen species than wild type. Therefore, in addition to functioning in the conversion of 7-HMC a to Chl a in senescent leaves, HCAR may play a critical role in protecting plants from high light-induced damage by preventing the accumulation of 7-HMC a and Pheo a in developing and mature leaves at the vegetative stage.