• Title/Summary/Keyword: C-N hydrolase

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Anti-lipid Peroxidative Principles from the Stem Bark of Kalopanax pictus Nakai

  • Choi, Jong-won;Han, Youn-Nam;Lee, Kyung-Tae;Park, Kun-Yong;Kwak, Tae-Soon;Kwon, Sang-Hyuk;Park, Hee-Juhn
    • Archives of Pharmacal Research
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    • v.24 no.6
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    • pp.536-540
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    • 2001
  • Hepatic lipid peroxide contents were examined in bromobenzene-treated rats firstly after the oral administration of MeOH extract of Kalopanax pictus stem bark its n-BuOH fraction, EtOAc fraction and an alkaline hydrolysate of the n-BuOH fraction, and secondly after the intraperitoneal administration of hederagenin monodesmosides and bisdesmosides. Two hederagenin monodesmosides, kalopanaxsaponin A (KPS-A) and sapindoside C, exhibited significant anti-lipid peroxidation effects after intraperitoneal administration at doses of $10-30{\;}{\mu}mole/kg$, whereas their bisdesmosides did not exhibit any significant activity. These results suggest that it is the hederagenin monodesmosides that are responsible for anti-lipid peroxidation in vivo. The activity of KPS-A was established by the observation of decreased aminopyrine N-demethylase activity and increased epoxide hydrolase activity.

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Anti-HIV and Antihepatotoxic Constituents from Medicinal Plant Resources

  • Park, Jong-Cheol;Park, Ju-Gwon;Hur, Jong-Moon;Hwang, Young-Hee;Jung, Deuk-Young
    • Plant Resources
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    • v.4 no.3
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    • pp.196-199
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    • 2001
  • Medicinal plants were screened for the inhibitory effects on human immunodeficiency virus type 1 pretense. Of the extracts tested, the strong inhibitory effects were observed in the acetone extracts of the pericarp of Camellia japonica. Camelliatannin H from the pericarp of C. japonica showed a potent inhibitory activity on HIV-1 pretense. Effects of the extract and compound from leaves of Zanthoxylum piperitum on the enzyme activities were investigated in the liver of bromobenzene-treated rats. The methanol extract and protocatechuic acid isolated from Z. pipetitum reduced the activity of aniline hydroxylase that increased by bromobenzene, while did not affect the activities of aminopyrin N-demethylase and glutathione S-transferase. The extract and protocatechuic acid recovered significantly the activity of epoxide hydrolase decreased by bromobenzene.

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Anti-HIV and Antihepatotoxic Constituents from Medicinal Plant Resources

  • Park, Jong-Cheol;Park, Ju-Gwon;Hur, Jong-Moon;Hwang, Young-Hee;Jung, Deuk-Young
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2001.11a
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    • pp.68-73
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    • 2001
  • Medicinal plants were screened for the inhibitory effects on human immunodeficiency virus type 1 pretense Of the extracts tested, the strong inhibitory effects were observed in the acetone extracts of the pericarp of Camellia japonica. Camelliatannin H from the pericarp of C. japonica showed a potent inhibitory activity on HIV- 1 pretense. Effects of the extract and compound from leaves of Zanthoxylum piperitum on the enzyme activities were investigated in the liver of bromobenzene-treated rats. The methanol extract and protocatechuic acid isolated from Z. piperitum reduced the activity of aniline hydroxylase that increased by bromobenzene, while did not affect the activities of aminopyrin N-demethylase and glutathione S-transferase The extract and protocatechuic acid recovered significantly the activity of epoxide hydrolase decreased by bromobenzene.

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Characterization of an Extracellular Xylanase in Paenibacillus sp. HY-8 Isolated from an Herbivorous Longicorn Beetle

  • Heo, Sun-Yeon;Kwak, Jang-Yul;Oh, Hyun-Woo;Park, Doo-Sang;Bae, Kyung-Sook;Shin, Dong-Ha;Park, Ho-Yong
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1753-1759
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    • 2006
  • Paenibacillus sp. HY-8 isolated from the digestive tracts of the longicorn beetle, Moechotypa diphysis, produced an extracellular endoxylanase with a molecular weight of 20 kDa estimated by SDS-PAGE. The xylanase was purified to near electrophoretic homogeneity from the culture supernatant after ammonium sulfate precipitation, gel filtration, and ionexchange chromatography. The purified xylanase exhibited the highest activities at pH 6.0 and $50^{\circ}C$. The $K_m\;and\;V_{max}$ values were 7.2 mg/ml and 16.3 U/mg, respectively, for birchwood xylan as the substrate. Nucleotide sequence of the PCR-cloned gene was determined to have the open reading frame encoding a polypeptide of 212 amino acids. The N-terminal amino acid sequence and the nucleotide sequence analyses predicted that the precursor xylanase contained a signal peptide composed of 28 amino acids and a catalytically active 19.9-kDa peptide fragment. The deduced amino acid sequence shared extensive similarity with those of the glycoside hydrolase family 11 of xylanases from other bacteria. The predicted amino acid sequence contained two glutamate residues, previously identified as essential and conserved for active sites in other xylanases of the glycoside hydrolase family 11.

A Novel Glycosyl Hydrolase Family 16 β-Agarase from the Agar-Utilizing Marine Bacterium Gilvimarinus agarilyticus JEA5: the First Molecular and Biochemical Characterization of Agarase in Genus Gilvimarinus

  • Lee, Youngdeuk;Jo, Eunyoung;Lee, Yeon-Ju;Hettiarachchi, Sachithra Amarin;Park, Gun-Hoo;Lee, Su-Jin;Heo, Soo-Jin;Kang, Do-Hyung;Oh, Chulhong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.776-783
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    • 2018
  • The agarase gene gaa16a was identified from a draft genome sequence of Gilvimarinus agarilyticus JEA5, an agar-utilizing marine bacterium. Recently, three agarase-producing bacteria, G. chinensis, G. polysaccharolyticus, and G. agarilyticus, in the genus Gilvimarinus were reported. However, there have been no reports of the molecular characteristics and biochemical properties of these agarases. In this study, we analyzed the molecular characteristics and biochemical properties of agarases in Gilvimarinus. Gaa16A comprised a 1,323-bp open reading frame encoding 441 amino acids. The predicted molecular mass and isoelectric point were 49 kDa and 4.9, respectively. The amino acid sequence of Gaa16A showed features typical of glycosyl hydrolase family 16 (GH16) ${\beta}$-agarases, including a GH16 domain, carbohydrate-binding region (RICIN domain), and signal peptide. Recombinant Gaa16A (excluding the signal peptide and carbohydrate-binding region, rGaa16A) was expressed as a fused protein with maltose-binding protein at its N-terminus in Escherichia coli. rGaa16A had maximum activity at $55^{\circ}C$ and pH 7.0 and 103 U/mg of specific activity in the presence of 2.5 mM $CaCl_2$. The enzyme hydrolyzed agarose to yield neoagarotetraose as the main product. This enzyme may be useful for industrial production of functional neoagaro-oligosaccharides.

Hepatoprotective Effect of the Methanol Fraction of Chinese Cabbage on Liver Injury in Rats Treated by bromobenzene (Chinese Cabbage 잎과 뿌리가 메탄올층의 Bromobenzene 간손상에 대한 보호효과)

  • Lee Hyo Jung;Kim Kwan Hyun;Lee Eun Ok;Choi Jong Won;Kang Kyung Sun;Yoon Byong Su;Kim Sung Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.5
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    • pp.1177-1181
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    • 2003
  • Chinese cabbage is a vegetable of Cruciferous family. It was usually consumed as Kimchi. It was known to have amount of vitamin c. Recently the trend for the development of functional food combined with oriental herbs. For this aim the study was performed to evaluate the hepatoprotective effect via antioxidant activity of leaf and root of Sanchon Chinese cabbage(Brassica campetris L.) comparatively. The methanol extracts of Chinese cabbage were tested for investigating the effects on the formation of lipid peroxide and the activities of free radical generating enzyme in vitro in bromobenzene-treated rats. The methanol extracts of chinese cabbage reduced bromobenzene-induced hepatic lipid peroxidation and inhibited the activity of xanthine oxidase. The methanol extracts of chinese cabbage did not activate amionopyrine N-demethylase, aniline hydroxylase and glutathione S-transferase. Epoxide hydrolase activity was decreased by bromobenzene, which was restored by pretreatment of the methanol extracts of chinese cabbage. The results suggest that the methanol extracts of Chinese cabbage is reduced by enhancing the activity of epoxide hydrolase.

A Novel Cellulase of the Mulberry Longicorn Beetle, Apriona germari, Dependent on N-Glycosylation for Enzymatic Activity

  • Lee, Seong-Jin;Kim, Seong-Ryul;Yoon, Hyung-Joo;Kim, IK-Soo;Lee, Kwang-Sik;Je, Yeon-Ho;Lee, Sang-Mong;Seo, Sook-Jae;Sohn, Hung-Dae;Jin, Byung-Rae
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.77-78
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    • 2003
  • A novel -1, 4-endoglucanase (EGase, EC 3.2.1.4) cDNA belonging to glycoside hydrolase family (GHF) 45 was cloned from the mulberry longicorn beetle, Apriona germari. The cDNA encoding EGase of A. germari (Ag-EGase) is 711 base pairs long with an open reading frame of 237 amino acid residues. The deduced protein sequence of Ag-EGase showed 54% and 48% identity to phytophagous beetle Phaedon cochleariae and termite Reticulitermes speratus hindgut symbiont, respectively. (omitted)

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Molecular Cloning and Sequence Analysis of Coelomic Cytolytic Factor-like Gene from the Midgut of the Earthworm, Eisenia Andrei (줄지렁이 중장에서 분리한 Coelomic cytolytic factor-유사 유전자의 클로닝 및 염기서열 분석에 관한 연구)

  • Baek, Nam Sook;Lee, Myung-Sik;Park, Sang-Kil;Kim, Dae-hwan;Tak, Eun-Sik;Ahn, Chi-Hyun;Sun, Zhenjun;Park, Soon Cheol
    • Journal of the Korea Organic Resources Recycling Association
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    • v.16 no.4
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    • pp.64-73
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    • 2008
  • The cDNA of CCF (coelomic cytolytic factor)-like gene (EC 3.2.1.16), a kind of glycosyl hydorlase, was isolated and cloned from the midgut of the earthworm Eisenia anderi. The size of nucleotide sequence appeared to be 1,152 bp and its predicted coding region was composed of 384 amino acid residues including the initiation methionine. The 17 residues at N-terminal end in the deduced amino acid sequence were regarded to be a signal peptide. Based on the amino acid sequence analysis, it appeared that this CCF-like protein could belong to glycosyl hydrolase family 16 (GHF16) and showed a high sequence homology of about 79~99% with CCF and CCF-like proteins from other earthworm species. The CCFs and CCF-like proteins from various earthworm species exhibited a 100% homology in the polysacchride-binding motif and glucanase motif. It has been reported that the CCFs isolated from E. fedita appeared to show a broader pattern recognition specificity than those from other earthworm species because this species resides in decaying organic matter showing very high microbial activity, implying that CCF-like protein isolated in this study from E. andrei might exhibit a broad substrate specificity that is a useful characteristic for industrial application. A phylogenetic analysis using the deduced amino acid sequences of CCF-related proteins through the BLASTX revealed that GHF16 families could be divided into three groups of metazoa, viriplantae and eubacteria subfamily. Subsequently the CCF-related proteins of metazoa subfamily could clearly be subgroup into lophotrochozoan and edysozoan type including a deuterostome origin. Further understanding of the biological properties of E. andrei CCF-like protein should be addressed to regulate the ${\beta}$-D-glucan hydrolysis and production for the industrial uses.

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Carboxy-Terminal Region of a Thermostable CITase from Thermoanaerobacter thermocopriae Has the Ability to Produce Long Isomaltooligosaccharides

  • Jeong, Woo Soo;Kim, Yu-Ri;Hong, Seong-Jin;Choi, Su-Jeong;Choi, Ji-Ho;Park, Shin-Young;Woo, Eui-Jeon;Kim, Young Min;Park, Bo-Ram
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1938-1946
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    • 2019
  • Isomaltooligosaccharides (IMOs) have good prebiotic effects, and long IMOs (LIMOs) with a degree of polymerization (DP) of 7 or above show improved effects. However, they are not yet commercially available, and require costly enzymes and processes for production. The N-terminal region of the thermostable Thermoanaerobacter thermocopriae cycloisomaltooligosaccharide glucanotransferase (TtCITase) shows cyclic isomaltooligosaccharide (CI)-producing activity owing to a catalytic domain of glycoside hydrolase (GH) family 66 and carbohydrate-binding module (CBM) 35. In the present study, we elucidated the activity of the C-terminal region of TtCITase (TtCITase-C; Met740-Phe1,559), including a CBM35-like region and the GH family 15 domain. The domain was successfully cloned, expressed, and purified as a single protein with a molecular mass of 115 kDa. TtCITase-C exhibited optimal activity at 40℃ and pH 5.5, and retained 100% activity at pH 5.5 after 18-h incubation. TtCITase-C synthesized α-1,6 glucosyl products with over seven degrees of polymerization (DP) by an α-1,6 glucosyl transfer reaction from maltopentaose, isomaltopentaose, or commercialized maltodextrins as substrates. These results indicate that TtCITase-C could be used for the production of α-1,6 glucosyl oligosaccharides with over DP7 (LIMOs) in a more cost-effective manner, without requiring cyclodextran.

Crystal Structure of Histidine Triad Nucleotide-Binding Protein from the Pathogenic Fungus Candida albicans

  • Jung, Ahjin;Yun, Ji-Sook;Kim, Shinae;Kim, Sang Ryong;Shin, Minsang;Cho, Dong Hyung;Choi, Kwang Shik;Chang, Jeong Ho
    • Molecules and Cells
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    • v.42 no.1
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    • pp.56-66
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    • 2019
  • Histidine triad nucleotide-binding protein (HINT) is a member of the histidine triad (HIT) superfamily, which has hydrolase activity owing to a histidine triad motif. The HIT superfamily can be divided to five classes with functions in galactose metabolism, DNA repair, and tumor suppression. HINTs are highly conserved from archaea to humans and function as tumor suppressors, translation regulators, and neuropathy inhibitors. Although the structures of HINT proteins from various species have been reported, limited structural information is available for fungal species. Here, to elucidate the structural features and functional diversity of HINTs, we determined the crystal structure of HINT from the pathogenic fungus Candida albicans (CaHINT) in complex with zinc ions at a resolution of $2.5{\AA}$. Based on structural comparisons, the monomer of CaHINT overlaid best with HINT protein from the protozoal species Leishmania major. Additionally, structural comparisons with human HINT revealed an additional helix at the C-terminus of CaHINT. Interestingly, the extended C-terminal helix interacted with the N-terminal loop (${\alpha}1-{\beta}1$) and with the ${\alpha}3$ helix, which appeared to stabilize the dimerization of CaHINT. In the C-terminal region, structural and sequence comparisons showed strong relationships among 19 diverse species from archea to humans, suggesting early separation in the course of evolution. Further studies are required to address the functional significance of variations in the C-terminal region. This structural analysis of CaHINT provided important insights into the molecular aspects of evolution within the HIT superfamily.