• Title/Summary/Keyword: {\alpha}$

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Biochemical Characterization of α-Galactosidase-Producing Thermophilic Bacillus coagulans KM-1 (α-Galactosidase를 생산하는 고온성 Bacillus coagulans KM-1 균주의 생화학적 특성)

  • Nam, Ki Ho;Jang, Mi Soon;Park, Hee Yeon;Koneva, Elena
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.5
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    • pp.516-521
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    • 2014
  • A bacterium producing ${\alpha}$-galactosidase (${\alpha}$-$\small{D}$-galactoside galactohydrolase, EC 3.2.1.22) was isolated. The isolate, KM-1 was identified as Bacillus coagulans based on its 16S rRNA sequence, morphology, and biochemical properties. ${\alpha}$-Galactosidase activity was detected the culture supernatant of B. coagulans KM-1. The bacterium showed the maximum activity for hydrolyzing para-nitrophenyl-${\alpha}$-$\small{D}$-galactopyranoside (pNP-${\alpha}Gal$) at pH 6.0 and $50^{\circ}C$. It hydrolyzed oligomeric substrates such as melibiose, raffinose, and stachyose liberating a galactose residue, indicating that the B. coagulans KM-1 ${\alpha}$-galactosidase hydrolyzed ${\alpha}$-1,6 linkage. The results suggest that the decreased stachyose and raffinose contents in fermented soybean meal are due to the ${\alpha}$-galactosidase activity.

Cloning and Characterization of ${\alpha}-Glucosidase$ Gene from Thermophilic Bacillus sp. DG0303

  • Lee, Yong-Eok
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.244-250
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    • 2000
  • An ${\alpha}-glucosidase$ gene (aglA) from thermophilic Bacillus sp. DG0303 was cloned, sequenced, and expressed in Escherichia coli. The aglA was localized to the 2.1-kb PvuI-XmnI region within the 5.9-kb DNA insert of the gybrid plasmid pAG1. The gene consisted of an open reading frame of 1,686 bp with an unusual GTG initiation codon and TGA termination codon. The amino acid sequence deduced from the nucleotide sequence predicted a protein of 562 amino acid residues with a M, of 66,551 dalton. A comparative amino acid sequence analysis revealed that DG0303 ${\alpha}-glucosidase$ is related to bacillary oligo-1, 6-glucosidases. The Bacillus sp. DG0303 ${\alpha}-glucosidase$ showed a high sequence identity (36-59%) to the B. flavocaldarius, B. cereus, and B. thermoglucosidasius oligo-1, 6-glucosidases. The number of prolines in theses four ${\alpha}-glucosidases. was observed to increase with increasing thermostability of these enzymes. The cloned ${\alpha}-glucosidase was purified from E. coli $DH5{\alpha}$ bearing pAG1 and characterized. The recombinant enzyme was identical with the native enzyme in its optimum pH and in its molecular mass, estimated by sodium dodecy1 sulfate-polyacrylamide gel electrophoresis. The temperature optimum of the cloned ${\alpha}-glucosidase$ was lower than that of the native enzyme.

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Purification and Application of Earthworm /alpha-Galactosidase by Affinity Chromatography (Galactomannan 이용에 관한 연구 : Affinity Chromatography에 의한 지렁이 유래 /alpha-Galactosidase의 정제 및 응용법)

  • 박귀근;정규훈;소림수행
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.298-303
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    • 1999
  • An $\alpha$-D-galactosidase ($\alpha$-D-galactoside galactohydrolase, EC 3. 2. 1. 22) from earthworm was purified by affinity chromatography using N-$\varepsilon$-aminocaproyl-$\alpha$-D-galactopyranosylamine coupled to sepharose and its properties were examined. The specific activity of the purified enzyme, tested with p-nitrophenyl-$\alpha$-D-galactopyranoside as substrate, was 314 units/mg protein, representing an 122-fold purification of the original crude extract. The final preparation obtained from by Sephadex G-25 chromatography showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 48,000 by SDS-polyacrylamide gel electrophoresis. The purified galactosidase was showed maximum activity at pH 4.5 and 4$0^{\circ}C$, and was stable in the pH and temperature ranges from 4.0 to 5.5 and 30 to 5$0^{\circ}C$, respectively. The enzyme activity was inhibited by Zn2+, Hg2+ and Co2+. When the purified $\alpha$-galactosidase treated to guar gum for 6 hour, gel-promoting property was increased. It was clear that enzymatic elimination of galactose from guar gum by purified $\alpha$-galactosidase would lead to a significant increase in gelation ability.

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The Effect of Betulinic Acid on $TNF-{\alpha}-induced$ MCP-1 Expression in HL-60 Cells (HL-60 세포에서 $TNF-{\alpha}$에 의한 MCP-1 발현에 미치는 Betulinic Acid의 효과)

  • Kim, Kyung-Chan;Lee, Chu-Hee
    • YAKHAK HOEJI
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    • v.52 no.1
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    • pp.37-42
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    • 2008
  • Betulinic acid, a naturally occurring pentacyclic triterpenoid, is found in abundance in the outer bark of white birch (Betula alba). In this study, we investigated if betulinic acid affects cytokine expression from activated macrophage cells. ELISA result showed that stimulation of HL-60 cells with proinflammatory cytokine such as $TNF-{\alpha}$ resulted in MCP-1 release into culture medium. In addition, transcriptional upregulation of MCP-1 in response to $TNF-{\alpha}$ was observed by RT-PCR analysis. However, incubation of HL-60 cells with betulinic acid prior to $TNF-{\alpha}$ treatment abrogated MCP-1 expression in transcription and translational level. Consistent with a number of studies which reported requirement of ERK activation for $TNF-{\alpha}$ expression, Western blot analysis showed that $TNF-{\alpha}-induced$ ERK activation was suppressed by pretreatment of HL-60 cells with betulinic acid. Taken together, our data indicate that betulinic acid exerts its anti-inflammatory effect through inhibition of $TNF-{\alpha}-induced$ ERK activation which is required for the subsequent MCP-1 release.

Photodegradation of Endosulfan alpha, beta, and sulfate in Aqueous Solution by UV Irradiation Only (자외선 조사에 의한 수용액 중에 함유되어 있는 엔도설판 alpha, beta, sulfate의 광분해)

  • Ryoo, Keon-Sang;Kim, Mi-Hyang;Lee, Hwa-Sung;Hwang, Hyun-Gick
    • Journal of Environmental Science International
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    • v.15 no.11
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    • pp.1061-1067
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    • 2006
  • Photodegradation of endosulfan alpha, beta, and sulfate known as the most toxic substance among organochlorine pesticides by UV irradiation was studied at experimental conditions such as different pH aqueous solution and reaction time. The initial concentration of endosulfan alpha, beta, and sulfate in aqueous solution was 500 ppb, respectively. The experiment of photodegradation was conducted in a quartz reactor equipped with a low pressure mercury lamp (100 W, 240 nm). The samples were withdrawn from the photo reactor at intervals of 0, 10 min, 30 min, 1 hr, 2 hr, and 4 hr. Endosulfan sulfate was never hydrolyzed and photodegraded in wide range of pH. At pH 5 and reaction time (240 min), endosulfan alpha was photodegraded up to 67%. Both endosulfan alpha and beta were started to photodegrade at pH 6.5 with the lapse of time, resulting in approximately 99.9% and 87.2% of photodegradation efficiency, respectively. Furthermore, at pH 9, endosulfan alpha and beta was partially hydrolyzed and photodegraded to 99.5% at 120 min of reaction time. During the photolysis, any photo-products of endosulfan alpha, beta, and sulfate were not observed.

Molecular Cloning of Estrogen Receptor $\alpha$ in the Masu Salmon, Oncorhynchus masou

  • Sohn, Young Chang
    • Journal of Aquaculture
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    • v.17 no.1
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    • pp.62-68
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    • 2004
  • A cDNA encoding the masu salmon, Oncorhynchus masou, estrogen receptor $\alpha$ (msER$\alpha$) was cloned from the pituitary gland by polymerase chain reaction (PCR). This cDNA contains an open reading frame encoding 513 amino acid residues, and the calculated molecular weight of this protein is about 56,430 Dalton. The amino acid sequences of the DNA binding and ligand binding domains of msER$\alpha$ showed high homology to those of other fish species (84-100%). Reverse transcription PCR analysis showed that the mRNA level of msER$\alpha$ in the pituitary was slightly higher in estradiol-17$\beta$(E2) injected masu salmon than that of control fish. To test the biological activity of msER$\alpha$, the cDNA was ligated to a mammalian expression vector and transfected into a gonadotrope-derived cell line, L$\beta$T2, with a reporter plasmid including estrogen responsive element. Expression of the reporter protein, luciferase, was E2 and msER$\alpha$-dependent. The masu salmon ER$\alpha$ is structurally conserved among teleost species and functions as a transcriptional activator in the pituitary cells.

The effects of buffer layer using $\alpha$-septithiophene on the organic light emitting diode (유기 전기 발광 소자에서 $\alpha$-septithiophene을 이용한 buffer layer의 영향)

  • Yi, Ki-Wook;Lim, Sung-Taek;Shin, Dong-Myung;Park, Jong-Wook;Park, Ho-Cheol
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2002.04b
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    • pp.53-56
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    • 2002
  • The effect of $\alpha$-septithiophene (${\alpha}-7T$) layers on the organic light emitting diode(OLED) was studied. The ${\alpha}-7T$ was used for a buffer layer in OLED. Hole injection was investigated and improved emission efficiency. The OLEDs structure can be described as indium tin oxide(ITO)/ buffer layer / hole transporting layer / emitting layer / electron transporting layer / LiF / Al. The hole transporting layer were composed of N,N-diphenyl-N,N-di(3-methylphenyl)-1,1-biphenyl-4,4-diamine(TPD), and N,N-di(naphthalene-1-ly)-N,N-diphenyl-benzidine( ${\alpha}$-NPD). The emitting layer, and electron transporting layer consist of tris(8-hydroxyquinolinato) aluminum($Alq_3$). All organic layer were deposited at a background pressure of less than $10^{-6}$ torr using ultra high vacuum (UHV) system. The ${\alpha}-7T$ layer can substitute the hole blocking layer, and improve hole injection properties.

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The Alpha Subunit of Go Interacts with Brain Specific High Mobility Group Box Containing Protein

  • Park, Jung-Sik;Ghil, Sung-Ho
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.405-411
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    • 2006
  • Heterotrimeric GTP binding proteins (G proteins) mediate signal transduction generated by neurotransmitter and hormones. Among G-proteins, Go is classified as a member of the Go/Gi family and the most abundant heterotrimeric G protein in brain. Most of the mechanistic analyses on the activation of Go indicated its action to be mediated by the $G{\beta}{\gamma}$ dimer because downstream effectors for its ${\alpha}$ subunit have not been clearly defined. To determine the downstream effectors of alpha subunits of Go ($Go{\alpha}$), we used yeast two-hybrid system to screen $Go{\alpha}$ interacting partners in cDNA library from the human brain. A brain specific high mobility group box containing protein (BHX), A possible transcription factor, was identified as a $Go{\alpha}$ interacting protein. We confirmed interaction between $Go{\alpha}$ and BHX employing in vitro affinity binding assay. Moreover, active form of $Go{\alpha}$ preferentially interacts with BHX than inactive farm. Our findings indicate that $Go{\alpha}$ could modulate gene expression via interaction with BHX during neuronal or brain development.

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Luteal Prostaglandin F2α: New Concepts of Prostaglandin F2α Secretion and Its Actions within the Bovine Corpus Luteuma - Review -

  • Okuda, K.;Skarzynski, D.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.3
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    • pp.390-400
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    • 2000
  • The corpus luteum (CL) is a temporary endocrine gland whose main function is to secrete progesterone to support pregnancy. On the other hand, the cyclic bovine CL has also been shown to be a site of prostaglandin $F_{2{\alpha}}$ ($PGF_{2{\alpha}}$) production. Although there is general agreement that endometrial $PGF_{2{\alpha}}$ is an essential luteolysin in cattle, luteal $PGF_{2{\alpha}}$ seems to play a luteotropic role as an autocrine and/or paracrine factor, especially for the development and maintenance of the CL. This supposition is based on evidence that some of the prerequisites for autocrine/paracrine mechanisms are present, including local production of $PGF_{2{\alpha}}$ and the existence of specific binding sites within the CL. The purpose of this paper is to review the regulation of luteal $PGF_{2{\alpha}}$ secretion, its action on CL as an autocrine and/or paracrine factor and the receptivity of bovine CL to. $PGF_{2{\alpha}}$.

The Formation of Hematite as Precursor for Magnetic Recording Media from Amorphous Ferric Hydroxide (비정질 수산화제이철로 부터 자기 기록 매체용 전구체인 헤마타이트 입자의 제조)

  • 변태봉;손진군;김태옥
    • Journal of the Korean Magnetics Society
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    • v.6 no.4
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    • pp.225-234
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    • 1996
  • Needle-like $\alpha-Fe_{2}O_{3}$ particles as precursor for magnetic recording media were prepared directly from amorphous ferric hydroxide in the aqueous solution by hydrothermal reaction. Ellipsoidal or rectangular $\alpha-Fe_{2}O_{3}$ particles were formed in the range of pH 10.75~11.75. The length and acicularity of $\alpha-Fe_{2}O_{3}$ particles were decreased gradually with increasing of citric acid concentration. The formation of needle-like $\alpha-Fe_{2}O_{3}$ particles was inhibited above citric acid concentration of $1.5{\times}10^{-4}\;mol$. We can synthesize $\alpha-Fe_{2}O_{3}$ particles with the most superior acicularity at $140^{\circ}C$ and can not expect a good needle-like particles above $220^{\circ}C$.

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