• Title/Summary/Keyword: -galactosidase

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Effect of Cyclic AMP on the Two Promoters of Escherichia coli Thioredoxin Gene

  • Sa, Jae-Hoon;Fuchs, James A.;Lim, Chang-Jin
    • BMB Reports
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    • v.30 no.5
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    • pp.320-325
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    • 1997
  • Thioredoxin is a multi-functional protein which is ubiquitous in microorganisms, animals and plants. Previously, expression of the E. coli thioredoxin gene was found to be negatively regulated by cAMP. In the present study, the effect of cAMP on two separate promoters of the E. coli thioredoxin gene was investigated. Cyclic AMP had a repressible effect on P1 and P1P2 promoter activity of the constructs. This effect was also observed in the cya strain. The P2 promoter construct gave very high -galactosidase activity, and its expression was not affected by exogenous cAMP. It was assumed that a cis-acting negative element, probably the cAMP-CRP binding site, might have been deleted in the P1 promoter construct. Repression of the thioredoxin gene expression by cAMP appeared to be independent of ppGpp.

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Partial Purification and Characterization of Thermostable Alkaline $\beta$-Mannanase from Bacillus sp. JB-99 Suitable for Pulp Bleaching

  • VIRUPAKSHI S.;BABU K. GlREESH;NAIK GAJANAN R.
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.689-693
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    • 2005
  • Bacillus sp. JB-99, when grown in a chemically defined medium containing lactose as a carbon source, yielded 3,860 U/ml extracellular $\beta$-mannanase, which was high compared to other examined carbon sources. Among the nitrogen sources, yeast extract enhanced the enzyme activity. The enzyme production was growth-associated. The enzyme was optimally active at $65^{\circ}C$, pH 10, and had a half-life of 190 min at $65^{\circ}C$. N-Bromosuccinamide and $AgNO_3,\;CuSO_4$, and $HgCl_2$ strongly inhibited the enzyme, whereas $Ca^{2+}$ stimulated the enzyme activity. The $\alpha$-galactosidase enzyme production was not found in any of the enzyme assays.

Bioconversion Using Lactic Acid Bacteria: Ginsenosides, GABA, and Phenolic Compounds

  • Lee, Na-Kyoung;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.869-877
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    • 2017
  • Lactic acid bacteria (LAB) are used as fermentation starters in vegetable and dairy products and influence the pH and flavors of foods. For many centuries, LAB have been used to manufacture fermented foods; therefore, they are generally regarded as safe. LAB produce various substances, such as lactic acid, ${\beta}$-glucosidase, and ${\beta}$-galactosidase, making them useful as fermentation starters. Existing functional substances have been assessed as fermentation substrates for better component bioavailability or other functions. Representative materials that were bioconverted using LAB have been reported and include minor ginsenosides, ${\gamma}$-aminobutyric acid, equol, aglycones, bioactive isoflavones, genistein, and daidzein, among others. Fermentation mainly involves polyphenol and polysaccharide substrates and is conducted using bacterial strains such as Streptococcus thermophilus, Lactobacillus plantarum, and Bifidobacterium sp. In this review, we summarize recent studies of bioconversion using LAB and discuss future directions for this field.

A plasmid vector faciliting gene expression in both yeast and mammalian cells

  • Lee, Tae-Ho
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.149-151
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    • 1997
  • A plasmid vector with combined features of yeast shuttle vector and mammalian expression vector was constructed to facilitate expression of cloned gene in both cell-types. All necessary elements required for plasmid maintenance and selection in E. coli, yeast and mammalian cells were size-economically arranged in this plasmid. The numan cytomegalovirus (CMV) immediate early promoter and yeast GAL1 promoter were sequentially placed in front of the gene to be expressed. The synthetic splicing donor and acceptor sequences were inserted into the immediate upstream and downstream of the GAL1 promotor, allowing the CMV promotor to direct the expression of a given gene in mammalian cell environment by splicing out the interfering GAL1 promotor sequence. When the resulting vector containing LacZ as a gene was introduced into yeast and mammalian cells, both cells efficiently produced .betha.-galactosidase, dimonstrating its dual host usage.

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Fnr, NarL and NarP Regulation and Time Course Expression of Escherichia coli aeg-46.5 Gene

  • Ahn, Ju-Hyuk;Choe, Mu-Hyeon
    • BMB Reports
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    • v.29 no.1
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    • pp.88-91
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    • 1996
  • The anaerobically expressed gene aeg-46.5, which had been identified by the operon fusion technique with a hybrid bacteriophage of ${\lambda}$ and Mu, ${\lambda}$placMu53, was studied for its expression pattern and growth. The expression of aeg-46.5 was studied in the wild-type cell and mutant cells that have mutation (s) in the control gene of anaerobic respiration (fnr) and nitrate response (narL and narP). The ${\beta}$-galactosidase reporter gene showed maximum expression in narL host after two hours of aerobic to anaerobic switch in M9-Glc-nitrate medium. Both 40 mM and 100 mM concentrations of nitrate ion in the medium had little effect on expression level. We propose that aeg-46.5 is subject to multiple regulations of anaerobic activation by Fnr, nitrate activation by NarP and repression mediated by NarL.

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Screening of Transcriptional Regulator of the Draf Proto-oncogene Using the Yeast One-hybrid System

  • Park, So-Young;Park, Na-Hyun;Kwon, Eun-Jeong;Yoo, Mi-Ye
    • Journal of Life Science
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    • v.9 no.2
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    • pp.52-56
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    • 1999
  • The Raf, a cytoplasmic serine/thereonine protein kinase, acts as an important mediator of signals involving cell proliferation, differentiation and development. Multiple regulatory elements should participate in the expression of D-raf, Drosophila homolog of human c-raf-1. In order to search regulatory factors involved in the D-raf promoter activation, we accomplished the yeast one-hybrid screening using D-raf promoter region from bp-330 to -309 with respect to the transcription initiation site as bait. After screening, sixteen independent positive clones of ${\beta}$-galactosidase activties were identified and sequenced. Two clones having 94-98% identity with daughterless and one clone having 93% identity with escargot by Blast search among these clones were screened.

Assay of Antimutagenic Activities of Vegetable Plants (식용식물(食用植物)의 항변이원성(抗變異原性)에 관한 연구(硏究))

  • Seo, Jung-Sook;Lee, Young-Wook;Suh, Nan-Joo;Chang, Il-Moo
    • Korean Journal of Pharmacognosy
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    • v.21 no.1
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    • pp.88-91
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    • 1990
  • Potential antimutagenic activities of vegetable plants were investigated. 24 vegetables which are frequently consumed by Korean people were extracted with 70% ethyl alcohol to prepare the extract samples. Then those samples were added to the culture media containing mitomycin C($O.3\;{\mu}g/ml$) and the SOS-Chromotest was performed. Positive control, mitomycin C alone, showed about 330 units of ${\beta}$-galactosidase activities. Among 24 vegetable samples, Saxifraga oblougifolia Nakai (취나물, Saxifragaceae) and Platycodon grandiflorum A. De Candolle(도라지, Campanulaceae) showed 136 and 155 units, respectively when mitomycin C was treated. These results indicate that Saxifragae Herba and Platycodi Radix possess protecting action from mutagenic activity produced by mitomycin C.

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Mutagenicity and Hepato-Toxicity of Kyoaesamultang (교애사물탕의 변이원성 및 간독성에 관한 연구)

  • 우덕안;홍희탁;문진영;이태균;김철호;김준기;최미정;남경수
    • Toxicological Research
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    • v.13 no.3
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    • pp.197-202
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    • 1997
  • Kyoaesamultang(KAT) has been used as an important prescription for various diseases including threatened abortion, associated with pregnancy in traditional medicine. In oder to identify the safety of KAT, this study was designed to determine mutagenicity and hepato-toxicity. In Rec-assay, Bacillus subtills H-17($Rec{^+}$) and M-45($Rec{^-}$) strains were used to clarify the DNA damage property. In Ames test, Salmonella typhimurium TA98 and TA100 were used for mutagenicity testing. In SOS umu test, Salmonella typhimurium TA1535 containing plasmid pSK1002 was used as a tester strain, and the levels of umu operon expression were monitored by measuring the $\beta$-galactosidase activity. From tested results, KAT did not show DNA damage and mutagenicity. On the other hand, hepato-toxicity of KAT to female ICR mice was monitored by the measurements of s-GOT, s-GPT and LDH activities after oral feeding for 15days. KAT showed 34% increase of s-GOT and s-GPT activities, also exhibited 35% increase of LDH activity in mice sera.

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Characterization of cell wall hydrolases induced by sugar starvation

  • Lee, Eun-Jeong;Koizumi, Nozomu
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.371-374
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    • 2005
  • In our previous work in transcriptional regulation of sugar, expression of genes encoding putative glycosyl hydrolases in Arabidopsis was induced by sugar starvation. They were annotated as b-galactosidase (At5g56870), ${\beta}-xylosidase$ (At5g49360) and ${\beta}-glucosidase$ (At3g60140), which belong to glycosyl hydrolase family that has a catalytic domain of polysaccharides. From the primary structure of deduced amino acid sequence, they were predicted to localize to cell wall. Further investigation of these cell wall hydrolases implicated that cell wall polysaccharides provide metabolizable sugars to nutrient allocation under sugar starvation.

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Studies on enzyme immunoassay for determining progesterone of bovine plasma and its clinical application I. Optimizing double antibody for progesterone in enzyme immunoassay (Enzyme immunoassay(EIA)에 의한 소의 혈중 progesterone 측정과 이의 응용에 관한 연구 I. 이항체(二項體)의 최적조건에 관한 연구)

  • Kang, Chung-boo;Shin, Jong-uk;Choe, Sang-yong
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.321-325
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    • 1988
  • This experiment was carried out to determine the progesterone concentration of bovine plasma by liquid phase double antibody enzyme immunoassay. The optimum conditions of assay-system, double (first and second) antibody and carrier (normal rabbit serum) were investigated. The optimum dilution rate of first antibody, second antibody and normal rabbit serum was $10{\times}10^3$ to $15{\times}10^3$, 20 and $1{\times}10^3$ times, respectively.

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