• Title/Summary/Keyword: Enzymatic activity

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Candidate of Probiotic Bacteria Isolated from Several Jeotgals: Korean Traditional Fermented Seafoods

  • Cho, Gyu-Sung;Do, Hyung-Ki;Bae, Chae-Yoon;Cho, Gyu-Sup;Whang, Cher-Won;Shin, Heuyn-Kil
    • Preventive Nutrition and Food Science
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    • v.11 no.2
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    • pp.140-145
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    • 2006
  • Seventy eight bacterial strains were isolated from several jeotgals using MRS and M 17 agar media. The probiotic properties such as tolerance of extreme growth condition, production of antimicrobial compound, production of hydrogen peroxide, and enzymatic activity of bile salt hydrolase were investigated. DHK 4, 10, 21 and 74 strains showed_a strong tolerance property against extreme conditions such as low pH and 0.5% oxgall-supplemented medium. DHK 10 and 47 strains produced hydrogen peroxide on TMB agar plate. DHK 8 and 10 strains produced antimicrobial compounds onto MRS agar against E. facalis. DHK 4, 6, 21, 29, 33, 63 and 87 strains had high activities of bile salt hydrolase. Especially, DHK 10 displayed a strong probiotic candidate; the abilities to produce the antimicrobial compound, hydrogen peroxide, and bile salt hydrolase. All these strains are assumed to be useful probiotic candidates. Among 78, twenty seven strains which have probiotic properties were tentatively identified by 16S rRNA sequencing. Among them, 7 Lactobacillus spp., 6 Leuconosotoc spp., 2 Weisella spp., 1 Pediococcus sp., 1 Staphylococcus sp., 1 Enterococcus sp. and 2 Streptococcus spp. were tentatively identified.

Studies on K-value and Nucleotides-Degrading Enzymes in Fish Muscle, - I (어육(魚肉)의 K치(値)와 핵산(核酸) 관련(關聯) 물질(物質)의 분해효소(分解酵素)에 관(關)한 연구(硏究)-I)

  • Kim, June- Pyung;Kim, Jong- Bae;Park, Il-Woong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.17 no.1
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    • pp.33-41
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    • 1988
  • Estimation of fish freshness is very important in food industries. it was estimated from the concentration of nucleotides. ammonia, amines and volatile acids in fish meat. Changes in the K-values of 12 species of fish were observed during storage at $0^{\circ}C$. The range of differences in the K-value-increasing rate of chilled fish muscle among fish species might be wider for white-fleshed fishes than for dark- fleshed fishes. As to the 12 species of fish examined. the coefficient of correlation between them was calculated to be 0.77. The fish muscle will he contained an effective inhibitor of IMP-degrading enzymatic activity.

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p38 Kinase Regulates Nitric Oxide-induced Dedifferentiation and Cyclooxygenase-2 Expression of Articular Chondrocytes

  • Yu, Seon-Mi;Cheong, Seon-Woo;Cho, Sam-Rae;Kim, Song-Ja
    • IMMUNE NETWORK
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    • v.6 no.3
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    • pp.117-122
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    • 2006
  • Background: Caveolin, a family of integral membrane proteins are a principal component of caveolae membranes. In this study, we investigated the effect of p38 kinase on differentiation and on inflammatory responses in sodium nitroprusside (SNP)-treated chondrocytes. Methods: Rabbit articular chondrocytes were prepared from cartilage slices of 2-week-old New Zealand white rabbits by enzymatic digestion. SNP was used as a nitric oxide (NO) donor. In this experiments measuring SNP dose response, primary chondrocytes were treated with various concentrations of SNP for 24h. The time course of the SNP response was determined by incubating cells with 1mM SNP for the indicated time period $(0{\sim}24h)$. The cyclooxygenase-2 (COX-2) and type II collagen expression levels were determined by immunoblot analysis, and prostaglandin $E_2\;(PGE_2)$ assay was used to measure the COX-2 activity. The tyrosine phosphorylation of caveolin-1 was determined by immunoblot analysis and immunostaining. Results: SNP treatment stimulated tyrosine phosphorylation of caveolin-1 and activation of p38 kinase. SNP additionally caused dedifferentiation and inflammatory response. We showed previously that SNP treatment stimulated activation of p38 kinase and ERK-1/-2. Inhibition of p38 kinase with SB203580 reduced caveolin-1 tyrosine phosphorylation and COX-2 expression but enhanced dedifferentiation, whereas inhibition of ERK with PD98059 did not affect caveolin-1 tyrosine phosphorylation levels, suggesting that ERK at least is not related to dedifferentiation and COX-2 expression through caveolin-1 tyrosine phosphorylation. Conclusion: Our results indicate that SNP in articular chondrocytes stimulates dedifferentiation and inflammatory response via p38 kinase signaling in association with caveolin-1 phosphorylation.

A Comparative Study on Spermatozoan Maturation in the Hibernating Animals (동면동물의 정자성숙과정에 대한 비교 연구)

  • Jae-Ho Chang;Yung-Keun Oh;In-Ho Choi;Noh-Pal Jung;Hyung-Cheul Shin
    • Biomedical Science Letters
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    • v.3 no.1
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    • pp.1-9
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    • 1997
  • Acrosome reaction usually occures just before fertilization in most mammals, and it has been known that $Ca^{2+}$ plays an important role in the acrosome reaction and albumin also known as a critical factor for spermatozoan activities. The present study has been designed in order to observe maturing processes of the spermatozoa occurred in the ductus epididymidis and to clarify the relationships of $Ca^{2+}$ concentrations with those processes, and to compare the enzymatic activities of ATPase and the lactate dehydrogenase of the spermatozoa in accordance with time before and after the spermatozoan maturation. From the results, we can confirm that most of the bat spermatozoa come to maturity within the epididymal cauda and may pass through capacitation outside the cauda. However it is expected to be studied that the fluctuation of spermatogenic activity depending on temperature changes and their relationships with the ductus epididymidis and their mutual influences.

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Enzymatic Decolorization of Various Dyes by Trametes versicolor KCTC 16781 (Trametes versicolor KCTC 16781에 의한 다양한 염료의 색도제거 특성)

  • 박철환;이유리;김탁현;이명구;이병환;이진원;김상용
    • KSBB Journal
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    • v.18 no.5
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    • pp.398-403
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    • 2003
  • Due to the low biodegradability of dyes, conventional biological wastewater treatment systems are inefficient in treating dye wastewater. Various white-rot fungi were investigated for the decolorization of six industrial dyes (reactive blue 5, reactive blue 16, reactive black 5, acid black 52, reactive orange 16, and acid violet 43). Among ten fungi, T. versicolor KCTC 16781 was selected as a testing strain because this had the best performance of decolorization for six dyes from the results of the solid culture experiments. In liquid culture medium, T. versicolor KCTC 16781 decolorized over 96% of six dyes for 48 hrs. Laccase started to produce in the early stage of the culture, and showed the highest peak value of 2.3 U/mL in 24 hrs. Enzyme activity remained constant until the end of culture. Fungal decolorization is a promising alternative to replace or supplement present treatment process.

Extracellular Secretion of a Maltogenic Amylase from Lactobacillus gasseri ATCC33323 in Lactococcus lactis MG1363 and its Application on the Production of Branched Maltooligosaccharides

  • Cho, Mee-Hyun;Park, Sang-Eun;Lee, Myung-Hun;Ha, Suk-Jin;Kim, Hae-Yeong;Kim, Myo-Jeong;Lee, Sung-Joon;Madsen, Soren M.;Park, Cheon-Seok
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1521-1526
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    • 2007
  • A maltogenic amylase gene from Lactobacillus gasseri ATCC33323 (LGMA) was expressed in Lactococcus lactis MG1363 using the P170 expression system. The successful production of recombinant LGMA (rLGMA) was confirmed by the catalytic activity of the enzyme in liquid and solid media. The N-terminal amino acid sequencing analysis of the rLGMA showed that it was Met-Gln-Leu-Ala-Ala-Leu-, which was the same as that of genuine protein, meaning the signal peptide was efficiently cleaved during secretion to the extracellular milieu. The optimal reaction temperature and pH of rLGMA ($55^{\circ}C$ and pH 5, respectively) and enzymatic hydrolysis patterns on various substrates (${\beta}$-cyclodextrin, starch, and pullulan) supported that rLGMA was not only efficiently secreted from the Lactococcus lactis MG1363 but was also functionally active. Finally, the branched maltooligosaccharides were effectively produced from liquefied com starch, by using rLGMA secreted from Lactococcus lactis, with a yield of 53.1%.

Regioselective Oxidation of Lauric Acid by CYP119, an Orphan Cytochrome P450 from Sulfolobus acidocaldarius

  • Lim, Young-Ran;Eun, Chang-Yong;Park, Hyoung-Goo;Han, Song-Hee;Han, Jung-Soo;Cho, Kyoung-Sang;Chun, Young-Jin;Kim, Dong-Hak
    • Journal of Microbiology and Biotechnology
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    • v.20 no.3
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    • pp.574-578
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    • 2010
  • Archaebacteria Sulfolobus acidocaldarius contains the highly thermophilic cytochrome P450 enzyme (CYP119). CYP119 possesses stable enzymatic activity at up to $85^{\circ}C$. However, this enzyme is still considered as an orphan P450 without known physiological function with endogenous or xenobiotic substrates. We characterized the regioselectivity of lauric acid by CYP119 using the auxiliary redox partner proteins putidaredoxin (Pd) and putidaredoxin reductase (PdR). Purified CYP119 protein showed a tight binding affinity to lauric acid ($K_d=1.1{\pm}0.1{\mu}M$) and dominantly hydroxylated (${\omega}-1$) position of lauric acid. We determined the steady-state kinetic parameters; $k_{cat}$ was 10.8 $min^{-1}$ and $K_m$, was 12 ${\mu}M$. The increased ratio to $\omega$-hydroxylated production of lauric acid catalyzed by CYP119 was observed with increase in the reaction temperature. These studies suggested that the regioselectivity of CYP119 provide the critical clue for the physiological enzyme function in this thermophilic archaebacteria. In addition, regioselectivity control of CYP119 without altering its thermostability can lead to the development of novel CYP119-based catalysts through protein engineering.

Characterization of a Squalene Synthase from the Thraustochytrid Microalga Aurantiochytrium sp. KRS101

  • Hong, Won-Kyung;Heo, Sun-Yeon;Park, Hye-Mi;Kim, Chul Ho;Sohn, Jung-Hoon;Kondo, Akihiko;Seo, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.759-765
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    • 2013
  • The gene encoding squalene synthase (SQS) of the lipid-producing heterotrophic microalga Aurantiochytrium sp. KRS101 was cloned and characterized. The krsSQS gene is 1,551 bp in length and has two exons and one intron. The open reading frame of the gene is 1,164 bp in length, yielding a polypeptide of 387 predicted amino acid residues with a molecular mass of 42.7 kDa. The deduced krsSQS sequence shares at least four conserved regions known to be required for SQS enzymatic activity in other species. The protein, tagged with $His_6$, was expressed into soluble form in Escherichia coli. The purified protein catalyzed the conversion of farnesyl diphosphate to squalene in the presence of NADPH and $Mg^{2+}$. This is the first report on the characterization of an SQS from a Thraustochytrid microalga.

Saccharification of Brown Macroalgae Using an Arsenal of Recombinant Alginate Lyases: Potential Application in the Biorefinery Process

  • Gimpel, Javier A.;Ravanal, Maria Cristina;Salazar, Oriana;Lienqueo, Maria Elena
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1671-1682
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    • 2018
  • Alginate lyases (endo and exo-lyases) are required for the degradation of alginate into its constituting monomers. Efficient bioethanol production and extraction of bioactives from brown algae requires intensive use of these enzymes. Nonetheless, there are few commercial alginate lyase preparations, and their costs make them unsuitable for large scale experiments. A recombinant expression protocol has been developed in this study for producing seven endo-lyases and three exo-lyases as soluble and highly active preparations. Saccharification of alginate using 21 different endo/exo-lyase combinations shows that there is complementary enzymatic activity between some of the endo/exo pairs. This is probably due to favorable matching of their substrate biases for the different glycosidic bonds in the alginate molecule. Therefore, selection of enzymes for the best saccharification results for a given biomass should be based on screens comprising both types of lyases. Additionally, different incubation temperatures, enzyme load ratios, and enzyme loading strategies were assessed using the best four enzyme combinations for treating Macrocystis pyrifera biomass. It was shown that $30^{\circ}C$ with a 1:3 endo/exo loading ratio was suitable for all four combinations. Moreover, simultaneous loading of endo-and exo-lyases at the beginning of the reaction allowed maximum alginate saccharification in half the time than when the exo-lyases were added sequentially.

Molecular Cloning and Expression of a Xylanase Gene from Thermophilic Alkalophilic Bacillus sp. K-17 in Escherichia coli (고온, 호알칼리성 Bacillus sp. K-17 Xylanase 유전자의 Escherichia coli 에의 클로닝 및 발현)

  • Sung, Nack-Kie;Chun, Hyo-Kon;Shim, Ki-Hwan;Kang, In-Soo;Teruhiko Akiba
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.178-182
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    • 1989
  • A gene coding for a xylanase of thermophilic alkalophilic Bacillus sp. K-17 was cloned in Escherichia coli C600 with pBR322. Plasmid pAXl13 was isolated from a transformant producing xylanase, and the xylanase gene was located in a 4.3 Kb HindIII fragment. Biotinylated pAXl13 hybridized to a 4.3 Kb HindIII fragment from chromosomal DNA of thermophilic alkalophilic Bacillus sp. K-17. The xylanase activity was observed in the extracellular curture fluid of E. coli carrying pAXl13. The pAXl13-encoded xylanase had the same enzymatic properties as those of xylanase I produced by thermophilic alkalophilic Bacillus sp. K-17.

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