• Title/Summary/Keyword: Methyl phosphate

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Lipase를 이용한 (S)-3-acetoxy-2-methylpropanol의 제조

  • 서영배;서연찬;이갑득
    • Microbiology and Biotechnology Letters
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    • v.24 no.2
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    • pp.213-216
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    • 1996
  • Optically active carboxylic acid, D-(-)-$\beta$-hydroxyisobutyric acid {(D)-(-)-HIBA} is a useful chiral starting material for the preparation of enantiomerically pure bioactive compounds which have a chiral methyl carbon center in the molecule such as captopril, $\alpha$-tocopherol, erythromycin A, muscone and so on. (S)-3-Acetoxy-2-methylpropanol can be used as the precursor of (D)-(-)-HIBA, that is, chemical oxidation of the hydroxyl group and subsequent hydrolysis of acyl group in (S)-3-acetoxy-2-methylpropanol affords D-(-)-$\beta$-hydroxyisobutyric acid. (S)-3-Acetoxy-2-methyl-propanol was prepared by lipase-catalyzed asymmetric hydrolysis. In the enzymatic hydrolysis system, lipase AY (Candida rugosa) provided the expected (S)-3-acetoxy-2-methylpropanol in 60% e.e. of the enantiomeric purity under the phosphate buffer and organic co-solvent system.

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Molecular cloning and expression analysis of the first two key genes through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway from Pyropia haitanensis (Bangiales, Rhodophyta)

  • Du, Yu;Guan, Jian;Xu, Ruijun;Liu, Xin;Shen, Weijie;Ma, Yafeng;He, Yuan;Shen, Songdong
    • ALGAE
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    • v.32 no.4
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    • pp.359-377
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    • 2017
  • Pyropia haitanensis (T. J. Chang et B. F. Zheng) N. Kikuchi et M. Miyata is one of the most commercially useful macroalgae cultivated in southeastern China. In red algae, the biosynthesis of terpenoids through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway can produce a direct influence on the synthesis of many biologically important metabolites. In this study, two genes of cDNAs, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and 1-deoxy-D-xylulose-5-phosphate reductase (DXR), which encoding the first two rate-limiting enzymes among MEP pathway were cloned from P. haitanensis. The cDNAs of P. haitanensis DXS (PhDXS) and DXR (PhDXR) both contained complete open reading frames encoding polypeptides of 764 and 426 amino acids residues, separately. The expression analysis showed that PhDXS was significant differently expressed between leafy thallus and conchocelis as PhDXR been non-significant. Additionally, expression of PhDXR and its downstream gene geranylgeranyl diphosphate synthase were both inhibited by fosmidomycin significantly. Meanwhile, we constructed types of phylogenetic trees through different algae and higher plants DXS and DXR encoding amino acid sequences, as a result we found tree clustering consequences basically in line with the "Cavalier-Smith endosymbiotic theory." Whereupon, we speculated that in red algae, there existed only complete MEP pathway to meet needs of terpenoids synthesis for themselves; Terpenoids synthesis of red algae derivatives through mevalonate pathway came from two or more times endosymbiosis of heterotrophic eukaryotic parasitifer. This study demonstrated that PhDXS and PhDXR could play significant roles in terpenoids biosynthesis at molecular levels. Meanwhile, as nuclear genes among MEP pathway, PhDXS and PhDXR could provide a new way of thinking to research the problem of chromalveolata biological evolution.

Phosphoryl Transferring Activity was Revealed from $F_1-ATPase$ of Escherichia coli by $^{31}P$ NMR Investigation

  • Sohn, Joon-Hyung;NamKung, Jun;Yoon, Joon-Ho;Woo, Mi-Kyoung;Yeh, Byung-Il;Choi, Jong-Whan;Kim, Hyun-Won
    • Biomedical Science Letters
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    • v.13 no.3
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    • pp.169-173
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    • 2007
  • [ $^{31}PNMR$ ] spectroscopy revealed the adenylate kinase-like activity and the phosphotransferase activity from $F_1-ATPase$ of Escherichia coli. Incubation of $F_1-ATPase$ with ADP in the presence of $Mg^{2+}$ shows the appearance of $^{31}P$ resonances from AMP and Pi, suggesting the generation of AMP and ATP by adenylate kinase-like activity and the subsequent hydrolysis to Pi. Incubation of $F_1-ATPase$ with ADP in the presence of methanol shows additional peak from methyl phosphate, suggesting phosphotransferase activity of $F_1-ATPase$. Both adenylate kinase-like activity and the phosphotransferase activity has not been reported from $F_1-ATPase$ from Escherichia coli. $^{31}P$ NMR proved that it could be a valuable tool for the investigation of phosphorous related enzyme.

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Optimal Production of Protease from Entomopathogenic Fungus Beauveria bassiana (곤충 병원성 곰팡이 Beauveria bassiana로부터 Protease의 최적 생산)

  • Ko, Hwi-Jin;Kim, Hyun-Kyu;Kang, Sun-Chul;Kwon, Suk-Tae
    • Applied Biological Chemistry
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    • v.39 no.6
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    • pp.449-454
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    • 1996
  • We investigated the optimal condition for the production of extracellular pretense(a cuticle-degrading pretense) from entomopathogenic fungus Beauveria bassiana(ATCC7159) in liquid medium by adding of gelatin, bovine serum albumin(BSA), casein and polypeptone. The optimal induction medium for production of extracellular pretenses is composed of 0.5% polypeptone, trace elements and 50 mM potassium phosphate(pH 6.0). In this condition, the production of extracellular pretenses increased rapidly after the 24hrs, peaking at the third day and there was little inductive effect in culture broth more than pH 7.0. The pretenses were inhibited by phenyl methyl sulfonyl fluoride(PMSF). High activity of pretense was showed both range of pH 8.5 and 11.5 and also detected by three different portions of slice gel derived from non-denaturing isoelectricfocusing gel. At least three different extracellular pretenses are produced in optimal production medium when polypeptone is used as the sole carbon and nitrogen source.

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A Study on the Adsorption Properties of Diisopropyl Methyl Phosphonate on Chromium-Based Metal-Organic Frameworks Using Acetic Acid as a Modulator (아세트산을 조절인자로 제작한 크롬 기반 금속유기골격체의 diisopropyl methyl phosphonate 흡착 특성 연구)

  • Sangjo Jeong
    • Applied Chemistry for Engineering
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    • v.34 no.6
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    • pp.596-602
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    • 2023
  • Chromium-based metal-organic frameworks (MIL-101(Cr)) were synthesized, and their potential use as a filling material for gas masks or protective clothing was assessed through adsorption experiments using diisopropyl methyl phosphate (DIMP) as a simulant for chemical warfare agents. MIL-101(Cr) was prepared using acetic acid (MIL-101(Cr)-A) and sodium hydroxide (MIL-101(Cr)-N) as modulators. The use of acetic acid as a modulator resulted in a larger specific surface area and a higher DIMP adsorption capacity. MIL-101(Cr)-A absorbed approximately 160% of its own weight of moisture when exposed to an environment with a relative humidity of 90% for 10 days, surpassing other adsorbents such as activated carbon. The DIMP adsorption capacity of MIL-101(Cr)-A decreased to about 40% of its initial adsorption capacity after 24 hours of exposure to an environment with a relative humidity of 90%. However, this capacity is still higher compared to that of activated carbon used in commercial gas masks, suggesting a high potential for future use as a filling material for gas masks or protective clothing.

Vitamin B6 Deficiency, Genome Instability and Cancer

  • Wu, Xia-Yu;Lu, Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5333-5338
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    • 2012
  • Vitamin B6 functions as a coenzyme in >140 enzymatic reactions involved in the metabolism of amino acids, carbohydrates, neurotransmitters, and lipids. It comprises a group of three related 3-hydroxy-2-methyl-pyrimidine derivatives: pyridoxine (PN), pyridoxal (PL), pyridoxamine (PM) and their phosphorylated derivatives [pyridoxal 5'-phosphate (PLP) and pyridoxamine 5'-phosphate (PMP)], In the folate metabolism pathway, PLP is a cofactor for the mitochondrial and cytoplasmic isozymes of serine hydroxymethyltransferase (SHMT2 and SHMT1), the P-protein of the glycine cleavage system, cystathionine ${\beta}$-synthase (CBS) and ${\gamma}$-cystathionase, and betaine hydroxymethyltransferase (BHMT), all of which contribute to homocysteine metabolism either through folate-mediated one-carbon metabolism or the transsulfuration pathway. Folate cofactors carry and chemically activate single carbons for the synthesis of purines, thymidylate and methionine. So the evidence indicates that vitamin B6 plays an important role in maintenance of the genome, epigenetic stability and homocysteine metabolism. This article focuses on studies of strand breaks, micronuclei, or chromosomal aberrations regarding protective effects of vitamin B6, and probes whether it is folate-mediated one-carbon metabolism or the transsulfuration pathway for vitamin B6 which plays critical roles in prevention of cancer and cardiovascular disease.

Metabolic engineering for isoprenoids production in Escherichia coli

  • Kim, Seon-Won;Keasling, J.D.
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.70-73
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    • 2001
  • Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all isoprenoids. IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of 1-deoxy-D-xylulose-5-phosphate(DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoismerase and encoded by dxr. To determine if one of more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains ($DH5{\alpha}$, XL1-Blue, and JM101) that had been engineered to produce lycopene, a kind of isoprenoids. Lycopene production was improved significantly in strains transformed with the dex expression vectors. At arabinose concentrations between 0 and 1.33 mM, cells expressiong both dxs and from $P_{BAD}$ on a midium-copy plasmid produced 1.4 -2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene production in cell expressing both dxs and dxr was lower than in cells expression dxs only. A comparison of the three E. coli strains trasfomed with the arabinose-inducible dxs on a medium-copy plasmid revealed that lycopene production was highest in XL1-Blue.

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Iontophoretic Transport of GHRP-6 (성장호르몬방출펩타이드-6 (GHRP-6)의 경피투과)

  • Choi, Bo-Kyung;Oh, Seaung-Youl
    • Journal of Pharmaceutical Investigation
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    • v.31 no.4
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    • pp.273-279
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    • 2001
  • The purpose of this study is to characterize the iontophoretic transport of growth hormone releasing peptides (GHRP-6) through hairless mouse skin from aqueous solution. The effect of various factors, such as pH, poloarity, current profile, current density, current duration, ionic strength, drug concentration, and enhancer application was studied to obtain basic knowledge on the transport. We have also studied the stability of GHRP-6 in solution with/without current. The donor chamber was filled with phosphate buffer solution containing GHRP-6 and the receptor chamber was filled with phosphate buffer solution (pH 7.4). Ag/AgCl electrode was used for their stability and reversibility. At a predetermined time interval, sampling was made and the concentration of drug was analysed using HPLC system. The results showed that, compared to passive flux, the total amount of drug transported increased markedly by the application of anodal current. Cathodal flux was similar to passive flux. Flux increased with the current density, the duration of current application and drug concentration. The effect of enhancers on the flux was studied using hydrophilic (5% N-methyl pyrrolidone) and hydrophobic (5% propylene glycol monolaurate, 5% oleic acid) enhancers. Application of enhancer also increased the flux.

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A STUDY ON THE CYTOTOXICITY OF THE ROOT CANAL SEALERS (근관충전용(根管充塡用) sealer의 세포독성(細胞毒性)에 관한 연구(硏究))

  • Lee, Seung-Jong;Kim, Yung-Hai
    • Restorative Dentistry and Endodontics
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    • v.16 no.1
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    • pp.25-40
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    • 1991
  • Four root canal sealers, Apatite Root Sealer I and II composed mainly of hydroxyapatite/tricalciumphosphate, Sealapex containing calcium hydroxide, and Roth Sealer composed of zinc oxide - eugenol were compared on the culture of L929 fibroblasts. MIT (Methyl Thiazole Tetrazolium Bromide) colorimetric technique was used to measure the mitochondrial dehydrogenase activity. Results were as follows: 1. Hydroxyapatite/tricalcium phosphate mixed sealers were significantly less toxic compared with calcium hydroxide and zinc oxide - eugenol type sealers. High pH of the calcium hydroxide sealer and release of eugenol component from the zinc oxide - eugenol type sealer were presumed to be the cause of the toxicity of these two sealers. In no cases, there were more cytoblastic effects in hydroxyapatite/tricalcium phosphate mixed sealers compared to the control groups. 2. In all experimental groups, toxicity was decreased as dilutions were increased. However in zinc oxide-eugenol type sealer the cell activity was weakened for all dilution groups. 3. Regarding the effect of setting time, Apatite I and Sealapex were less toxic as the setting progressed. Apatite II kept constant regardless of the different time ellapsed after setting but Roth sealer revealed significantly higher toxicity for all experimental groups. 4. Comparing two different culture periods of 24 hours and 72 hours, Apatite I showed higher cell activities in longer period(72 hours) while Apatite II did not. Sealapex and Roth sealer, however, showed significantly lower cell activities in longer period.

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Effect of Garlic Oil on Fatty Acid Accumulation and Glycerol-3-Phosphate Dehydrogenase Activity in Differentiating Adipocytes

  • He, M.L.;Yang, W.Z.;You, J.S.;Chaves, A.V.;Mir, P.S.;Benchaar, C.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.12
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    • pp.1686-1692
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    • 2009
  • Garlic oil (GAR, Allium sativum L.) has been studied as a feed additive to improve animal production performance and decrease methane emission in ruminants. The present study was designed to determine the possible effect of GAR on fatty acid composition and accumulation in animal fat tissue using a cell model. 3T3-L1 preadipocytes at $2{\times}10^{4}\;mL^{-1}$ were seeded to 24-well plates and allowed to proliferate to reach confluence. The cells were then treated with media containing 0, 2.5, 5, 10, 20 and 40 $\mu{g}$ $mL^{-1}$ of GAR during the differentiation period for 8 days. Media containing dexamethasone, methyl-isobutylxanthine and insulin was applied during the first 2 days of the early differentiation period. On day 8 sub-sets of the wells were stained with oil red-O and the remaining cells were harvested for determination of glycerol-3-phosphate dehydrogenase [EC 1.1.1.8] (GPDH) activity (n = 6) and cellular fatty acid concentration (n = 6). It was found that supplementation of GAR increased (p<0.05) the ratio of monounsaturated fatty acids/saturated fatty acids in the adipocytes and showed inhibitory effect (p<0.05) on the post-confluent proliferation. With relative low dosage, GAR (5-20 $\mu{g}$ $mL^{-1}$) increased (p<0.05) the GPDH activity without affecting the cellular fatty acid concentration, while a high dosage (40 $\mu{g}$ $mL^{-1}$) inhibited (p<0.05) fatty acid accumulation and decreased GPDH activity. Supplementation of GAR had an effect on cell post-confluent proliferation, differentiation and fatty acid accumulation. However, the effect may be diverse and depends on the dose applied.