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Hanseniaspora thailandica BC9 β-Glucosidase for the Production of β-ᴅ-Hexyl Glucoside

  • Phongprathet, Sujittra;Vichitphan, Kanit;Han, Jaehong;Vichitphan, Sukanda;Sawaengkaew, Jutaporn
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.579-587
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    • 2018
  • For biotechnological production of high-valued ${\beta}-{\text\tiny{D}}$-hexyl glucoside, the catalytic properties of Hanseniaspora thailandica BC9 ${\beta}$-glucosidase purified from the periplasmic fraction were studied, and the transglycosylation activity for the production of ${\beta}-{\text\tiny{D}}$-hexyl glucoside was optimized. The constitutive BC9 ${\beta}$-glucosidase exhibited maximum specific activity at pH 6.0 and $40^{\circ}C$, and the activity of BC9 ${\beta}$-glucosidase was not significantly inhibited by various metal ions. BC9 ${\beta}$-glucosidase did not show a significant activity of cellobiose hydrolysis, but the activity was rather enhanced in the presence of sucrose and medium-chain alcohols. BC9 ${\beta}$-glucosidase exhibited enhanced production of ${\beta}-{\text\tiny{D}}$-hexyl glucoside in the presence of DMSO, and 62% of ${\beta}-{\text\tiny{D}}$-hexyl glucoside conversion was recorded in 4 h in the presence of 5% 1-hexanol and 15% DMSO.

Synthesis and Secretion of the Endo-$\beta$-l,4-Glucanase from Bacillus subtilis in Industrial Yeast Strain (산업용 효모에서 Bacillus subtilis Endo-$\beta$-1,4-Glucanase의 생합성 및 분비)

  • 박용준;이영호;백운화;강현삼
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.348-355
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    • 1991
  • DNA segment encoding $\beta$-1, 4-glucanase of Bacillus subtilis was fused in frame to mouse $\alpha$-amylase signal sequence behind the alcohol dehydrogenase isoenzyme I gene (ADHI) promoter of the yeast expression vector pMS12. To enhance the expression level of the $\beta$glucanase gene in yeast, transcription terminator sequence iso-1-cytochrome c gene (CYCI) was inserted into the recombinant plasmid. The transformants harbouring such recombinant plasmids secreted $\beta$-glucanase into the culture medium. The expresstion level of the $\beta$-glucanase gene was increased about 2-fold caused by inserting the terminator. The amount of the secreted $\beta$-glucanase in culture medium was approximately 60% of the total quantity synthesized.

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Effect of Dietary Level of $\beta$-Carotene on Antioxidants Contents and Hepatic Morphology in Rats ($\beta$-Carotene 의 수준별 공급에 따른 흰주의 항산화 물질과 간조직의 형태적 변화)

  • 최은미;박정룡;서정숙
    • Journal of the East Asian Society of Dietary Life
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    • v.3 no.2
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    • pp.51-61
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    • 1993
  • This study was designed to investigate the effect of dietary level of $\beta$-carotene on level of antioxidant nutrients of rat tissues. Male Sprague Dawley rats at the age of 30days were fed on diets containing different levels of $\beta$-carotene(0, 10, 120, 1200, 12000mg per kg diet). Body weight gain of rats fed with 12000mg $\beta$-carotene diet was significantly decreased, but liver and heart weights were not significantly different among groups, The content of total glutathione tended to decrease significantly in 12000mg $\beta$-carotene diet group when compared to $\beta$-carotene restriction group(BC O). However, total vitamin C content of liver showed the tendency to increase by $\beta$ -carotene supply up to 1200mg. But this tendency was not found in plasma, The content of zinc in liver and plasma was significantly decreased by $\beta$-carotene restriction. Alkaline phosphatase activity was significantly higher in 12000mg diet group. In case of $\beta$-carotene restriction group, fibroblasts were proliferated in portal endothelium, and the vacuolar size was enlarged more than the nuclear, In 12000mg diet group, hepatic vacuoles were extended, but their size was regular and the lysis of hepatocytes was observed. Also, fibroblasts were proliferated in portal endothelium and the regular vacuolar size was extended.

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Identification of Mutations in Protein Kinase CKIIβ Subunit That Affect Its Binding to Ribosomal Protein L41 and Homodimerization

  • Ahn, Bong-Hyun;Lee, Ji-Hoon;Bae, Young-Seuk
    • BMB Reports
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    • v.36 no.4
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    • pp.344-348
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    • 2003
  • Protein kinase CKII is composed of two catalytic ($\alpha$ or $\alpha$') subunits and two regulatory ($\beta$) subunits. The $CKII{\beta}$ subunit is thought to mediate the tetramer formation and interact with other target proteins. However, its physiological function remains obscure. In this study, point mutants of $CKII{\beta}$ that are defective for the L41 binding were isolated by using the reverse two-hybrid system. A sequence analysis of the point mutants revealed that Asp-26, Met-52, and Met-78 of $CKII{\beta}$ are critical for L41 binding; Asn-67 (and/or Lys-139) and Met-52 are important for $CKII{\beta}$ homodimerization. Two point mutants, R75 and R83, of $CKII{\beta}$ interacted with L5, topoisomerase $II{\beta}$, and CKBBP1/SAG, but not with the wild-type $CKII{\beta}$. This indicates that $CKII{\beta}$ homodimerization is not a prerequisite for its binding to target proteins. These $CKII{\beta}$ point mutants may be useful in exploring the biochemical physiological functions of $CKII{\beta}$.

Production of Sialytrisaccharides Using $\beta$-Galactosidase and trans-Sialidase in One Pot

  • Lee, Sun-Gu;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.3
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    • pp.215-218
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    • 2000
  • Sialytrisaccharides based on $\beta$-galactosyldisaccharides were synthesized using $\beta$-galactosidase and trans-sialidase in one pot. Using $\beta$-galactosidase from Bacillus Ciculans and trans-sialidase from Trypanosoma cruzi simulaneously, 6mM sialyltrisaccharides composed of about 95% NeuAc$\alpha$(2,3)Gal$\beta$(1,4)GlcNAc and 5% NeuAc$\alpha$(2,3)Gal$\beta$(1,6)GlcNAc were produced from a reaction mixture containing 25mM o-nitropheny1-$\beta$-D-galsctolneuraminic acid. One beauty of this reaction was that a secondary hydrolysis of the disaccharide intermediate occurring between the activated galactopyranoside and N-acetylgucosamine was prevented. Using $\beta$-galactosidase from Escherichia cloi and the same trans-sialidase, 15mM sialyltrisaccharides composed of about 90% NeuAc$\alpha$(2,3)Gal$\beta$(1,6)GlcNac and 10% NeuAc$\alpha$(2,3)Gal$\beta$(1,4)GlcNAc were produced from a reaction misture containing 400nM galactose, 800nM N-acetylglucosylation rection between galactose and N-actylgucosamine was diminant since the disaccharide intermediate mainly resulted sreulted in the silylated product.

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Synergistic Effect of Lipopolysaccharide and Interferon-$\beta$ on the Expression of Chemokine Mig mRNA

  • Lee, Moon-Sook;Kim, Sung-Kwang;Kim, Hee-Sun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.813-818
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    • 2002
  • Expression of monokine induced by IFN-$\gamma$(Mig) mRNA is well-known to strictly depend on Interferon-$\gamma$(IFN-$\gamma$). Lipopolysaccharide (LPS) alone Is weakly effective on Mig mRNA expression in mouse Peritoneal macrophages. This study was undertaken to investigate the synergistic effect of LPS and IFN-$\beta$ on chemokine Mig gene expression in mouse peritoneal macrophages. Although IFN-$\beta$ alone was minimally effective, LPS plus IFN-$\beta$ synergized to produce a high level of Mig mRNh. The synergistic effect of LPS and IFN-$\beta$ (LPS/IFN-$\beta$) on Mig mRNA expression was strain-specific. The most effective synergistic effect of LPS/IFN-$\beta$ on the mRNh expression was found in simultaneous stimulation of LPS/IFN-$\beta$. This synergy was modulated at the level of the gene transcription and was not dependent on a new protein synthesis. Synergistic effect of LPS/IFN-$\beta$ also required the activation of $NF-_KB$. Accordingly, these data suggest that LPS/IFN-$\beta$ synergizes the expression of Mig mRNA through a process that depends on a pretranscriptional level and/or coincident Mig mRNA transcription.

Mercury induced the Accumulation of Amyloid Beta (Aβ) in PC12 Cells: The Role of Production and Degradation of Aβ

  • Song, Ji-Won;Choi, Byung-Sun
    • Toxicological Research
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    • v.29 no.4
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    • pp.235-240
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    • 2013
  • Extracellular accumulation of amyloid beta protein ($A{\beta}$) plays a central role in Alzheimer's disease (AD). Some metals, such as copper, lead, and aluminum can affect the $A{\beta}$ accumulation in the brain. However, the effect of mercury on $A{\beta}$ accumulation in the brain is not clear. Thus, this study was proposed to estimate whether mercury concentration affects $A{\beta}$ accumulation in PC12 cells. We treated 10, 100, and 1000 nM $HgCl_2$ (Hg) or $CH_3HgCl_2$ (MeHg) for 48 hr in PC12 cells. After treatment, $A{\beta}_{40}$ in culture medium increased in a dose- and time-dependent manner. Hg and MeHg increased amyloid precursor protein (APP), which is related to $A{\beta}$ production. Neprilysin (NEP) levels in PC12 cells were decreased by Hg and MeHg treatment. These results suggested that Hg induced $A{\beta}$ accumulation through APP overproduction and reduction of NEP.

In Vitro Characterization of Protein Kinase CKII β Mutants Defective in β-β Dimerization

  • Kim, Tae-Hyun;Lee, Jae-Yong;Kang, Beom Sik;Bae, Young-Seuk
    • Molecules and Cells
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    • v.19 no.1
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    • pp.124-130
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    • 2005
  • Protein kinase CKII is composed of two catalytic (${\alpha}$ or ${\alpha}^{\prime}$) subunits and two regulatory (${\beta}$) subunits. The ${\beta}$ subunit mediates tetramer formation through ${\beta}-{\beta}$ homodimerization and ${\alpha}-{\beta}$ heterodimerization. In a previous study R26 and R75, point mutants of $CKII{\beta}$ defective in ${\beta}-{\beta}$ dimerization, were isolated. In the present work we characterized these $CKII{\beta}$ mutants in vitro. Purified R26 and R75 bound to $CKII{\alpha}$ but were defective in binding to $CKII{\beta}$. R75 stimulated the catalytic activity of CKII whereas R26 gave little stimulation, and poly-L-lysine increased the stimulation of catalytic activity by R26 or R75. Circular dichroism and intrinsic fluorescence data pointed to different conformational changes in R26 and R75. Molecular modeling of these mutants provides an explanation of the difference in their ability to interact with $CKII{\beta}$ and to activate $CKII{\alpha}$.

Characteristics and Activity Changes of $\beta$-Galactosidase during Maturation and Postharvest of Persimmon Fruits (감과실의 성숙과 추숙중의 $\beta$-Galactosidase활성 변화 및 특성)

  • 신승렬;하유덕;김진구;김순동;김광수
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.6
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    • pp.605-611
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    • 1990
  • $\beta$-Galactosidase activity was not detected at green mature stage but were 21.79 and 380.23 units/100g-fr. wt. in mature and soft persimmon, respectively. The molecular weight of $\beta$-galactosidase was estimated to be 115, 000 daltons by the method of gel filtration. Vmax and Km value were 0.095m mo1e p-nitrophenyl-galactoside and 1.8$\times$10$^{-2}$ mM, respectively. The optimum temperature and pH of $\beta$-galactosidase were 45$^{\circ}C$ and 4.2, respectively. $\beta$-Galactosidase was inhibited by SDS.

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$18{\beta}$-Glycyrrhetinic Acid Induces Protective Anti-Candida albicans Antibody by Its Immunoadjuvant Activity ($18{\beta}$-Glycyrrhetinic Acid의 면역보조제효능에 의한 항 전신성캔디다증 효과)

  • Han, Yong-Moon
    • YAKHAK HOEJI
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    • v.52 no.6
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    • pp.494-499
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    • 2008
  • The role of antibody in the fungal infections is controversial. However, our previous reports showed a certain epitope in Candida albicans cell wall (CACW) induces protective antibody. A major problem is that the epitope isolation requires tremendous time with high cost. This aspect led us to investigate a simple way inducing protective antibodies against C. albicans. In the present study, we determined if $18{\beta}$-glycyrrhetinic acid ($18{\beta}$-GA) from Glabrae Radix (a family of Leguminosae) has immunoadjuvant activity. Data displayed that the $18{\beta}$-GA suppressed proliferations of both T- and Blymphocytes at high concentrations, whereas below 20 ${\mu}M$ concentration the compound supported the proliferations. These observations indicate that $18{\beta}$-GA has immunoregulatory activity. Based on this observation, an immunoadjuvant effect was examined at the low concentration. Results from animal experiments showed that CACW combined with or without $18{\beta}$-GA produced the anti-C. albicans antiserum in mice. Nevertheless, the CACW combined with $18{\beta}$-GA formula only protected mice against disseminated candidiasis (P<0.05). These data implicate that $18{\beta}$-GA has immunoadjuvant activity, which may provoke the CACW antigen to induce protective antibody. Currently, we are investigating possible mechanism of how the $18{\beta}$-GA provokes such protective immunity against the disseminated disease.