• Title/Summary/Keyword: TEX>-galactosidase

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Changes on the Components of Free Polysaccharide from Cell Wall of Persimmon Fruit by Treatments of Cell Wall Degrading Enzymes (세포벽분해효소의 처리에 따른 감과실의 세포벽 유리 다당류의 변화)

  • 신승렬;김미현
    • Food Science and Preservation
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    • v.2 no.1
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    • pp.173-183
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    • 1995
  • This paper was carried out to investigate changes in chromatograms of polysacctatides and soluble pectins on Sephadex G-50 and non-cellulosic neutral sugars of polysaccharides isolated from cell wall of persimmon fruits treated with polygalacturonase and $\beta$-galactosidase in vitro. The chromatogram pattern of soluble pectins extracted from cell wall treated with $\beta$-galactosidase on Sephacryl S-500 column were similar to those of untreatment, but contents of soluble pectins treated with $\beta$-galactosidase were different from those of untreatment. The patterns of chromatograms In soluble pectins extracted from cell wall treated with polygalacturonase were more complex and lower molecular polymer than those of other cell wall-degrading enzyme treatments. Non-cellulosic neutral sugar of polysaccharides in fraction I of soluble material treated with polygalacturonase was rhamnose, those in fraction II were similar to those in fraction III and contents of arabinose, xylose and glucose were higher than contents of other non-cellulosic neutral sugars. Non-cellulosic neutral sugars of polysaccharides in fraction I in soluble material by $\beta$-galactosidase treatment were rhamnose, arabinose, galactose and mannose. Content of glucose of polysaccharides in fraction II was higher than that in fraction I . Non-cellulosic neutral sugars treated with mixed enzyme were rhamnose, fucose, arabinose, xylose, mannose, galactose and glucose. Compositions of non-cellulosic neutral sugars of polysaccharides in fraction I were similar to those in fraction II and III.

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Studies on the Production of $\beta$-Galactosidase by Lactobacillus sporogenes - Production of Extracellular $\beta$- Galactosidase - (Lectobacillus sporogenes에 의한 $\beta$-Galactosidase 생산에 관한 연구 ( I ) -균체외 $\beta$-Galactosidase의 생산 -)

  • 김영만;이정치;정필근;최용진;양한철
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.59-66
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    • 1983
  • Cultural conditions for the production of extracellar $\beta$-galactosidase by Loctobacillus sporogenes, a spore forming lactic acid bacterium, were investigated with shaken flask and jar fermenter cultures. The fermentation medium giving maximum $\beta$-galactosidase yield was found to consist of 1 % lactose as a carbon source, 1.5% peptone as an organic nitrogen source. 0.2% ammonium sulfate as an inorganic nitrogen source, 0.8% ammonium phosphate dibasic as a phosphorus source, and 0.05% potassium chloride and 0.001% ferric chloride as mineral source. Optimal initial pH of the medium was 7.0 and the highest enzyme excretion was observed after 40 hours of cultivation at 37$^{\circ}C$. In this experiment, the 500$m\ell$ conical flask containing 50-200$m\ell$ of medium was shaken at 140 strokes per minute with 7cm amplitude in a reciprocating shaker. The maximum enzyme value attained was 38 U/$m\ell$ of the culture broth which was found to be slightly higher than the highest intermolecular enzyme activity (30 U/$m\ell$) observed after 24 hours of incubation. In the fermentor culture, the fermentation profile was shown to be similar to that observed in the shaken flask experiment. But the maximum extracellular enzyme activity was 45 U/$m\ell$ to be even higher than the value obtained with the shaken flask culture.

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Characteristics of ${\beta}-galactosidase$ activity in Lactobacillus plantarum from kimchi (김치에서 분리한 유산균 Lactobacillus plantarum의 ${\beta}-galactosidase$ 특성에 관한 연구)

  • Kang, Me-Seon;Rhee, Young-Hwan
    • Applied Biological Chemistry
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    • v.39 no.1
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    • pp.60-66
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    • 1996
  • To investigate a possible application of three strains of lactic acid bacteria(strain No. 49. No. 61. No. 75) from kimchi in milk fermentation industry, the optimal condition for production of intracellular ${\beta}-galactosidase$ from Lactobacillus(L.) plantarum and its enzymatic properties were examined. The preferable carbon source of the medium for strain No. 49 in production of ${\beta}-galactosidase$ was MRS broth with 1.0% lactose instead of dextrose of pH 65. for strain No. 75 with 1.0% galactose and for strain No. 61 with 3.0% lactose at pH 7.5, respectively. The maximum enzyme production from strain No. 49, No. 75 was observed after 48 hours culture at $30^{\circ}C$ in a medium containing the appropriate carbon source, from strain No. 61 after 48 hours culture at room temperature. The optimum temperature for ${\beta}-galactosidase$ activity from L. plantarum was $60^{\circ}C$ for strain No. 49, $37^{\circ}C$ for strain No. 61 and $50^{\circ}C$ for strain No. 75, respectively. The heat stability of enzyme activities for all three strains remained 90% at $45^{\circ}C$. The optimal pH was pH 6.5 and enzyme activities were most stable at pH for all three bacteria.

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Isolation of Bacillus sp. Producing ${\beta}-Galactosidase$ with High Transgalactosylation Activity and its Culture Characteristics Regarding Enzyme Production (갈락토스 전이활성이 높은 ${\beta}-galactosidase$ 생산균의 분리 및 효소생산과 관련된 몇가지 특징)

  • Kim, Min-Hong;Jung, Jin;In, Man-Jin
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.502-506
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    • 1995
  • A Bacillus strain which produces ${\beta}-galactosidase$ with high transgalactosylation activity, was isolated from soil and tentatively designated as Bacillus sp. A1. When ${\beta}-galactosidase$ from Bacillus sp. A1 reacted with 40% (w/w) lactose, transgalactosylation ratio reached up to 90% at the 70% conversion of the initial lactose. The biosynthesis of the enzyme in Bacillus sp. A1 required lactose as an inducer and was repressed by glucose. Observing that the addition of amino acids to culture medium resulted in enhancing, to a significant extent, both the growth and the enzyme production of the strain, yeast extract and commercially available hydrolysates of protein were examined for the suitability as amino acid source. As it turned out, SMP, an enzymatic hydrolysis product of soybean protein from Fuji Oil Co.(Japan), was the most suitable for optimization of the culture medium. When Bacillus sp. A1 was cultured in the presence of 0.5% SMP and 2% lactose, the enzyme activity increased up to $1.8\;U/m{\ell}-broth$.

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The Effects on Escherichia coli and Plasmid DNA Using Ultrasoft X-ray (Ultrasoft X-ray의 Escherichia coli균과 plasmid DNA에 대한 영향)

  • ;;;;;Seiya Chiba;Atsuo Kimura
    • KSBB Journal
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    • v.15 no.1
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    • pp.84-87
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    • 2000
  • We studied the effect of ultrasoft X-ray obtained from the Pohang Light Source (PLS), on the mutation of E. coli and the damage of plasmid. After irradiation, the supercoiled plasmid DNA converted to the relaxed-form, and then to the linear-form. We transformed the irradiated plasmid DNA and isolated $\beta$-galactosidase mutants. We also isolated $\beta$-galactosidase mutants from the directly irradiated cells. There were preferred mutational sites on DNA induced by ultrasoft X-ray.

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(${\beta}-Galactosidase$와 isoflavone의 미세캡슐화와 in vitro에서의 비배당화에 관한 연구)

  • Kim, Nam-Cheol;Jeon, Byeong-Ju;Gwak, Hae-Su
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2005.05a
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    • pp.322-325
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    • 2005
  • 본 연구는 ${\beta}-galactosidase$에 의한 isoflavone 배당체의 비배당화 연구를 실용화하기 위하여 바람직하지 않은 관능적 요소와 화학적 반응을 제어하기 위하여 두 물질을 미세캡슐화 하고 그것의 in vitro 안정성을 연구하여 기능성식품개발에 응용토록 하는데 목적을 두었다. 실험 결과, 미세캡슐화 수율이 매우 높았으며, 인공위액 상태에서의 안정성은 높은 반면에, 인공 소장액 상태에서의 안정성이 매우 낮았다. 또한 인공소장액에서 유리된 ${\beta}-galactosidase$에 의한 유리 isoflavone의 비배당화율이 약 75% 정도로 관찰되었다. 결과적으로 판단해 볼 때, isoflavone과 ${\beta}-galactosidase$의 미세캡슐화를 통한 소화기 내에서의 비배당화와 흡수율을 극대화하는데 매우 긍정적인 가능성을 보였다.

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Expression of Schizosaccharomyces pombe Thioltransferase and Thioredoxin Genes under Limited Growth Conditions

  • Cho, Young-Wook;Sa, Jae-Hoon;Park, Eun-Hee;Lim, Chang-Jin
    • BMB Reports
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    • v.34 no.5
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    • pp.395-401
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    • 2001
  • Schizosaccharomyces pombe gene encoding redox enzymes, such as thioltransferase (TTase) and thioredoxin (TRX), were previously cloned and induced by oxidative stress. In this investigation, their expressions were examined using $\beta$-galactosidase fusion plasmids. The expression of the two cloned genes appeared to be growth-dependent. The synthesis of $\beta$-galactosidase from the TTase-lacZ fusion was increased in the medium with the low glucose level, whereas it was significantly decreased in the medium without glucose or with galactose. It was also decreased in the nitrogen-limited medium. The synthesis of galactosidase from the TRX-lacZ fusion was unaffected by galactose or low glucose. However, it was lowered the absence of glucose. The synthesis of $\beta$-galactosidase from the TTase-lacZ fusion was shown to be enhanced in a higher medium pH. Our findings indicate that S. pombe TTase and TRX genes may be regulated by carbon and nitrogen sources, as well as medium pH.

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Production of Oligosaccharide by Beta-galactosidase of Kluyveromycces fragilis (Kluyveromyces fragilis의 Beta- galactosidase에 의한 Oligosaccharide의 생성)

  • 강국희;김상희
    • Journal of Food Hygiene and Safety
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    • v.2 no.2
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    • pp.67-73
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    • 1987
  • Kluyveromyces fragilis로부터 $\beta$-galactosidase의 생성조건과 조효소액의 성질 그리고 당 전이 반응에 의한 oligosaccharide 생성을 조사한 결과 다음과 같은 결과를 얻었다. \circled1 Peptone-Yeast extract 배지에 6% lactose를 첨가하였을 때 최대 효소생산을 보였다. \circled2 0.05 M potassium phosphate buffer (pH 7.0)에서 3% toluene를 첨가하여 37$^{\circ}C$ 5시간 배양하였을 때 K. fragilis로부터 효소가 최대로 추출되었다. \circled3 효소의 최적온도는 4$0^{\circ}C$이고 4$0^{\circ}C$ 이상의 열처리에서는 효소가 파괴되었으며, pH6-7에서는 상당히 안정하였다. \circled4 ONPG 기질로 사용하였을 때 Km 값은 2.5mM이었다. \circled5 당 전이 반응의 결과, 7개의 oligosaccharide가 생성되었다. 이상의 실험결과로 볼 때, 본 실험에 사용한 K. fragilis SKD 7001은 Beta-galactosidase의 생산을 위해서 이용 가치가 인정되었으며, 특히, 이 효소의 활성이 중성 pH에서 강하고 안정한 상태를 보이는 것은 시유나 원료우유의 lactose를 중성에서 해야 한다는 점을 고려할 때, 실용적 가치가 있다고 본다. 또, 이 효소의 작용과정에서 생성되는 oligosacchride는 장내 bifidus균의 생육을 촉진시키는 효과가 인정되고 있기 때문에 이용가치를 더욱 높여주는 것으로 생각된다.

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Transfer of Foreign Gene into Mud Loach, Misgurnus mizolepis I . Availability of the lacZ as a reporter gene for producing transgenic mud loach (미꾸라지, Misgurnus mizozepis에 외래 유전자 이식 I. lacZ의 reporter 유전자로서의 유용성 검토)

  • KIM Dong Soo;NAM Yoon Kwon
    • Journal of Aquaculture
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    • v.7 no.1
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    • pp.41-54
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    • 1994
  • In order to evaluate the availability of lacZ as a reporter gene for producing transgenic mud loach, foreign DNA, bacterial \beta-galactosidase$ gene (lacZ) was microinjected into mud loach eggs and its insertion and expression were examined. X-gal based histochemical assay, fluorimetric analysis of \beta-galactosidase$ with 4-methylumbelliferyl-$\beta$-D-galactoside (MUG) and molecular biological examination using polymerase chain reaction (PCR), dot blot, southern blot and sequence analysis of PCR products were carried out to analyze both microinjected group and non-injected controls. The results are disccussed in this paper.

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Characterization of the nar Promoter of Escherichia coli to use as an inducible promoter in Wild-type host Agrobacterium.tumefaciens

  • Lee, Gil-Ho;Jo, Mu-Hwan;Lee, Jong-Won
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.758-761
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    • 2001
  • In this study, the nar promoter of E. coli was characterized to see whether the nar promoter cloned onto pBBR122 can be used as an expression promoter of gram negative microbes. For this purpose, a plasmid with lacZ gene expressing ${\beta}-galactosidase$ instead of the structural genes of nar operon in a gram negative host strain(Agrobacterium.tumefaciens) was used to simplify an assay of induction of the nar promoter. The following effects were investigated to find optimal conditions: methods of inducing the nar promoter, optimal nitrate concentration, maximally inducing the nar promoter, the amount of expressed ${\beta}-galactosidase$ and induction ratio(specific ${\beta}-galactosidase$ activity after maximal induction/specific ${\beta}-galactosidase$ activity before induction). The following results were obtained from the experiments: the growth of Agrobacterium with E.coli nar promoter was not much affected by nitrate concentration in the shake-flask; induction of nar promoter was optimal when Agrobacterium was grown in the presence of 1% nitrate ion at the beginning of culture and when overnight culture was completely grown in the shake-flask before being transferred to other shake-flask; the amount of ${\beta}-galactosidase$ per cell and per medium volume was maximal when Agrobacterium was grown under aerobic condition to $OD_{600}$ of 1.7; then the nar promoter was induced under microaerobic and anaerobic condition made by lowering dissolved oxygen level(DO). After 2-3h of induction in the YEP medium selected as a main culture medium, the specific ${\beta}-galactosidase$ activity became about 17,000 Miller units in the fermentor cluture.

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