• Title/Summary/Keyword: Esterase activity

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Changes in Textural Properties of Korean Radish and relevant Chemical, Enzymatic Activities during Salting (염장과정 중 무의 조직감과 이와 관련된 화학적, 효소활성 변화)

  • Rhee, Hee-Seoup;Lee, Gui-Ju
    • Journal of the Korean Society of Food Culture
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    • v.8 no.3
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    • pp.267-274
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    • 1993
  • This study was aimed to investigate the changes in textural properties of Korean radish and relevant chemical, enzymatic activities during salting. During salting, pH was decreased and total acidity was increased. The maximum compression and puncture forces of Korean radish were decreased significantly whereas cutting force was increased. From the force-distance curves, the break point and maximum force point disappeared in salted Korean radish whereas these appeared apparently in fresh one. Also, the number of peak obtained by three types of test from salted Korean radish was decreased. Hot water soluble pectin and 0.4% Na-hexametaphosphate soluble pectin were increased whereas 0.05 N-HCl soluble pectin were decreased significantly. Polygalacturonase activity were increased in Korean radish solid(RS) and Korean radish juice(RJ) until 4 days of salting. Pectin esterase activity were decreased in RS and RJ. Cx-cellulase activity did not appear initially, however, they began to show their activities after 2 days of salting and were increased in RJ although it was low.

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Native and Foreign Proteins Secreted by the Cupriavidus metallidurans Type II System and an Alternative Mechanism

  • Xu, Houjuan;Denny, Timothy P.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.791-807
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    • 2017
  • The type II secretion system (T2SS), which transports selected periplasmic proteins across the outer membrane, has rarely been studied in nonpathogens or in organisms classified as Betaproteobacteria. Therefore, we studied Cupriavidus metallidurans (Cme), a facultative chemilithoautotroph. Gel analysis of extracellular proteins revealed no remarkable differences between the wild type and the T2SS mutants. However, enzyme assays revealed that native extracellular alkaline phosphatase is a T2SS substrate, because activity was 10-fold greater for the wild type than a T2SS mutant. In Cme engineered to produce three Ralstonia solanacearum (Rso) exoenzymes, at least 95% of their total activities were extracellular, but unexpectedly high percentages of these exoenzymes remained extracellular in T2SS mutants cultured in rich broth. These conditions appear to permit an alternative secretion process, because neither cell lysis nor periplasmic leakage was observed when Cme produced a Pectobacterium carotovorum exoenzyme, and wild-type Cme cultured in minimal medium secreted 98% of Rso polygalacturonase, but 92% of this exoenzyme remained intracellular in T2SS mutants. We concluded that Cme has a functional T2SS despite lacking any abundant native T2SS substrates. The efficient secretion of three foreign exoenzymes by Cme is remarkable, but so too is the indication of an alternative secretion process in rich culture conditions. When not transiting the T2SS, we suggest that Rso exoenzymes are probably selectively packaged into outer membrane vesicles. Phylogenetic analysis of T2SS proteins supports the existence of at least three T2SS subfamilies, and we propose that Cme, as a representative of the Betaproteobacteria, could become a new useful model system for studying T2SS substrate specificity.

Effects of a New 1,25(OH)$_2$-Vitamin $D_3$ Anglog on Proliferation and Differentiation of the Human Histiocytic Lymphoma Cell Line U937 (인체 Histiocytic Lymphoma Cell Line U937의 종식 및 분화에 대한 새로운 $1.25(OH)_2D_3$ 유도체의 효과에 관한 연구)

  • Jung, Soo-Ja;Suh, Myung-Ja;Rhu, Beung-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.3
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    • pp.443-452
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    • 1994
  • This study describes the effects of novel1, 25-dihydroxyvitamin D$_3$ analong[1,25(OH)$_2$-16ene-23yne-26, 27-F6-D$_3$] on proliferation of the human histiocytic lymphoma cell line U937 in vitro. We also examined the expression of c-myc oncogene in U937 cells was apparently inhibited to 62% and 87% of the control level after 4 days in the presence of 10-8M and 10-7 M of this analog, respectively. This compound morpholgically and functionally differentiated U937 cells to nonocyte-macrophage phenotype showing the increase of adherence ability to surface and a decrease of N/C ratio in Giemsa staining . Especially, nonspecific esterase activity which is a marker of cell differentiation to monocyte-macrophage was positive, and production of the positive stained cells increased in a dose dependent fashion . The expression of c-myc oncogene by 1, 25(OH)$_2$D$_3$ analog(10-7 M) was reduced by 60% at the mRNA level as determined by Northern blotting. The effects of this novel analog on cell proliferation and cell differentiation may open op new therapeutic strategies for human disorders such as psoriassis and may provide a tool to understand the mechanism of action of vitamin D$_3$ seco-steroids in malignancy.

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The Effect of Bee Venom on Scopolamine Induced Memorial Impairment (봉약침액(蜂藥鍼液)이 Scopolamine으로 유발(誘發)된 기억(記憶) 장애(障碍)에 미치는 영향(影響))

  • Song, Jeong-Yeon;Song, Ho-Sueb
    • Journal of Acupuncture Research
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    • v.23 no.3
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    • pp.103-115
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    • 2006
  • Alzheimer's disease (AD) is the most prevalent form of neurodegenerative disease associated with aging in the human population. This disease is characterized by the following 4 structural changes : Atrophy of the Cortex, Parasympathetic, and other neural cells, the existence of Neurofibrillary tangles (NFTs), and the accumulation of Senile plaques. NFTs and Senile plaques is known to be the index of this disease. Senile plaques disturbs the neutro transmission and depletes of Acetylcholine. So, Recovery of Acetylcholine is the primal objective for treating Alzheimer's disease. So, Inhibiting the activity of Acetylcholine Esterase (AChE), which causes the hydrolysus of acetylcholine into choline and acetate, can be seen as a key role for treating Alzheimer's disease. Increasing body of evidence has been demonstrated that Bee Venom Acupuncture (BV) could compete with complex protein involving in multiple step of $NF-_{\kappa}B$ activation and exert the anti -inflammatory potential of combined inhibition of the prostanoid and nitric oxide synthesis systems by inhibition of IKK and $NF-_{\kappa}B$. The effect of BV through behavioral tests after memory impairment induced by Scopolamine. We examined the improving effect of BV on the Scopolamine (1 mg/Kg, i.p.)-induced memorial impairment using passive avoidance response and water maze tests in the mice. BV (0.84, $1.67\;{\mu}g/ml$) reversed the Scopolamine-induced memorial impairment in dose dependent manner. This study therefore suggests that BV acupuncture method may be useful for prevention of development or progression of AD.

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Characterization of the pcbD Gene Encoding 2-Hydroxy-6-Ox0-6-Phenylgexa-2,4-Dienoate Hydrolase from Pseudomonas sp. P20

  • Lim, Jong-Chul;Lee, Jeong-Rai;Lim, Jai-Yun;Min, Kyung-Rak;Kim, Chi-Kyung;Ki, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.258-263
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    • 2000
  • 2-Hydroxy-6-oxo-6phenylhexa-2,4-dienoate (HOPDA) hydrolase catalyzes the hydrolytic cleavage of HOPDA to bemzpate and 2-hydroxypenta-2, 4-dienoate (HPD) during microbial catabolism of biphenyl and polychlorinated biphenyls. A HOPDA hydrolase gene (pcbD) was isolated from the genomic library of Pseudomonas sp. P20 and designated as pCNUO1201; a 7.5-kb XbaI DNA fragment from Pseudomonas sp. P20 was inserted into the pBluescript SK(+) XbaI site. E. coli HB101 harboring pCNU1201 exhibited HOPDA hydrolase activity. The open reading frame (ORF) corresponding to the pcbD gene consisted of 855 base pairs with an ATG initiation codon and a TGA termination codon. The ORF was preceded by a rebosome-binding sequence of 5'-TGGAGC-3' and its G+C content was 55 mol%. The pcbD gene of Pseudomonas sp. P20 was located immedeately downstream of the pcbC gene encoding 2,3- dihydroxybiphenyl 1,2-dioxygenase, and approximately 4-kb upstream of the pcbE gene encoding HPD hydratase. The pcbK gene was able to encode a polypeptide with a molecular weight of 31,732 containing 284 amino acid residues. The deduced amino acid sequence of the HOPDA hydrolase of Pseudomonas sp. P20 exhibited high identity (62%) with those of the HOPDA hydrolases of P. putida KF715, P. pseudoalcaligenes KF707, and Burkholderia cepacia LB400, and also significant homology with those of other hydrolytic enzymes including esterase, transferase, and peptidase.

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Applicability of Fluorescein Diacetate (FDA) and Calcein-AM to Determine the Viability of Marine Plankton (FDA와 Calcein-AM 방법을 이용한 해양플랑크톤 생사판별기법)

  • Baek, Seung-Ho;Shin, Kyoung-Soon
    • Ocean and Polar Research
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    • v.31 no.4
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    • pp.349-357
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    • 2009
  • Ballast water is widely recognized as a serious environmental problem due to the risk of introducing non-indigenous aquatic species. In this study we aimed to investigate measures which can minimize the transfer of aquatic organisms from ballast water. Securing more reliable technologies to determine the viability of aquatic organisms is an important initiative in ballast water management systems. To evaluate the viability of marine phytoplankton, we designed the staining methods of fluorescein diacetate (FDA) and Calcein-AM assay on each target species belonging to different groups, such as bacillariphyceae, dinophyceae, raphidophyceae, chrysophyceae, haptophyceae and chlorophyceae. The FDA method, which is based on measurements of cell esterase activity using a fluorimetric stain, was the best dye for determining live cells of almost all phytoplankton species, except several diatoms tested in this study. On the other hand, although fluorescence of Calcein-AM was very clear for a comparatively longer time, green fluorescence per cell volume was lacking in most of the tested species. According to the Flow CAM method, which is a continuous imaging technique designed to characterize particles, green fluorescence values of stained cells by FDA were significantly higher than those of Calcein-AM treatments and control, implying that the Flow CAM using FDA assay could be adapted as an important tool for distinguishing living cells from dead cells. Our results suggest that the FDA and Calcein-AM methods can be adapted for use on phytoplankton, though species-specific characters are greatly different from one organism to another.

Characterization of a Soil Metagenome-Derived Gene Encoding Wax Ester Synthase

  • Kim, Nam Hee;Park, Ji-Hye;Chung, Eunsook;So, Hyun-Ah;Lee, Myung Hwan;Kim, Jin-Cheol;Hwang, Eul Chul;Lee, Seon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.248-254
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    • 2016
  • A soil metagenome contains the genomes of all microbes included in a soil sample, including those that cannot be cultured. In this study, soil metagenome libraries were searched for microbial genes exhibiting lipolytic activity and those involved in potential lipid metabolism that could yield valuable products in microorganisms. One of the subclones derived from the original fosmid clone, pELP120, was selected for further analysis. A subclone spanning a 3.3 kb DNA fragment was found to encode for lipase/esterase and contained an additional partial open reading frame encoding a wax ester synthase (WES) motif. Consequently, both pELP120 and the full length of the gene potentially encoding WES were sequenced. To determine if the wes gene encoded a functioning WES protein that produced wax esters, gas chromatography-mass spectroscopy was conducted using ethyl acetate extract from an Escherichia coli strain that expressed the wes gene and was grown with hexadecanol. The ethyl acetate extract from this E. coli strain did indeed produce wax ester compounds of various carbon-chain lengths. DNA sequence analysis of the full-length gene revealed that the gene cluster may be derived from a member of Proteobacteria, whereas the clone does not contain any clear phylogenetic markers. These results suggest that the wes gene discovered in this study encodes a functional protein in E. coli and produces wax esters through a heterologous expression system.

Degradation of Diazinon and Dursban in Submerged Soil (담수양중(湛水壤中) Diazinon 과 Dursban 의 분해(分解)에 관(關)하여)

  • Choi, Jong-Woo;Lee, Kyu-Seung
    • Korean Journal of Environmental Agriculture
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    • v.6 no.2
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    • pp.1-11
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    • 1987
  • The degradation of two chemicals seem to be clearly affected by soil microbial activity in submerged soil $conditions(30{\pm}1^{\circ}C)$. The Active ingredient of Diazinon disappeared about 5 times faster than that of Dursban. By Applying 300% higher concentrations of both chemicals. under the above soil conditions, however, degradation was retarded by about one day. Some of the metabolites of Diazinon were as follows: 0.0-diethyl phosphorothioate and sulfotep as hydrolytic products, and diazoxon, 0.0-diethyl-0-[2-(1-hydroxy-1, 1-dimethyl)-6-methyl]-pyrimidinyl phosphorothioate and 2-isopropyl-6-methyl-pyrimidine-4-one as degradation products of monooxygenase. But 0. 0-diethyl phosphorothioate was the only methabolite of Dursban.

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Changes in Pectic Enzyme Activity during Maturation of Tomato Fruit and Purification, Some Properties of Polygalacturonase Isozymes (Tomato 성숙중(成熟中) Pectin 효소(酵素)의 활성변화(活性變化)와 Polygalacturonase의 분리(分離) 및 그 성질(性質))

  • Suh, Cha Hwan;Sohn, Tae Hwa;Choi, Jong Uck;Choi, Sang Won
    • Current Research on Agriculture and Life Sciences
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    • v.4
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    • pp.42-49
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    • 1986
  • The changes in pectinesterase (PE) and polygalacturonase (PG) activities were investigated during maturation and ripening of tomato fruits. PG was isolated from ripe tomatoes and purified by using DEAE-sephadex A-50 column chromatography. Its property was examined by general methods. PE activity was increased during maturation and then peaked at the on set of color change. PG activity was not detectable in mature green tomato fruits but increased during softening. Slab electrophoresis of the crude enzyme isolated from ripe tomatoes shoed 6 bands. Two polygalacturonase (PGI, PGII) were separated from crude enzyme of ripe tomatoes by chromatography on DEAE-sephadex A-50. PGI and PGII were purified by 61 fold and 98 fold by the present procedure, respectively. The Rm values of partially purified PGI and PGII were estimated to be 0.25 and 0.31, respectively. The optimum temp, and pH of PGI activity were $37^{\circ}C$ and pH 4.5, while those of PGII activity were $40^{\circ}C$ and pH 4.7, respectively. Isozyme PGI activity was the most stable at pH 4.3 and retained 50% of its activity when exposed at $78^{\circ}C$ for 5 min.

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Ginsenoside Rh2 inhibits proliferation of human promyelocytic HL-60 leukemia cells via $G_0/G_1$ phase arrest and induction of differentiation

  • Cho, Seoung-Hee;Kim, Dong-Hyun;Lee, Kyung-Tae
    • Proceedings of the Ginseng society Conference
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    • 2006.05a
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    • pp.3-12
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    • 2006
  • 1 The present work was performed to investigate the effects of ginsenoside Rh2 on proliferation, cell cycle-regulation and differentiation of human leukemia HL-60 cells as well as the underlying mechanisms for these effects. 2 Ginsenoside Rh2 potently inhibited the proliferation of HL-60 cells in both a dose- and time-dependent manner with an $IC_{50}$, $20{\mu}M$. 3 DNA flow-cytometry indicated that ginsenoside Rh2 markedly induced a $G_1$ phase arrest of HL-60 cells. 4 Among the $G_1$ phase cell cycle-related proteins, the levels of cyclin-dependent kinase(CDK)4, 6 and cyclin D1, cyclin D2, cyclin D3 were reduced by ginsenoside Rh2, whereas the steadystate levels of CDK2 and cyclin E were unaffected. 5 The protein levels of a CDK inhibitor p16, $p21^{CIP1/WAF1}$ and $p27^{KIP1}$ were markedly increased by ginsenoside Rh2. 6 Ginsenoside Rh2 markedly enhanced the binding of $p21^{CIP1/WAF1}$ and $p27^{KIP1}$ with CDK2 and CDK6, resulting in the reduced activity of both kinases and the hypophosphorylation of Rb protein. 7 We furthermore suggest that ginsenoside Rh2 is a potent inducer of the differentiation of HL-60 cells, based on observations such as a reduction of the nitroblue tetrazolium level, an increase in the esterase activities and phagocytic activity, morphology changes, and the expression of CD11b, CD14, CD64 and CD66b surface antigens. 8 In conclusion, the onset of ginsenoside Rh2-induced the $G_0/G_1$ arrest of HL-60 cells prior to the differentiation is linked to a sharp up-regulation of the $p21^{CIP1/WAF1}$ level and a decrease in the CDK2, CDK4 and CDK6 activities. This is the first report demonstrating that ginsenoside Rh2 potently inhibits the proliferation of human promyelocytic HL-60 cells via the $G_1$ phase cell cycle arrest and differentiation induction.

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