簡易檢索 / 詳目顯示

研究生: 李嘉修
Chia-hsiu Lee
論文名稱: 酵素催化合成肉桂酸衍生物
Enzymatic synthesis of Cinnamic Acid Derivatives
指導教授: 朱義旭
Yi-Hsu Ju
口試委員: 陳志平
none
陳秀美
none
學位類別: 碩士
Master
系所名稱: 工程學院 - 化學工程系
Department of Chemical Engineering
論文出版年: 2005
畢業學年度: 93
語文別: 英文
論文頁數: 55
中文關鍵詞: 自由基抗氧化
外文關鍵詞: free radical, antioxidant
相關次數: 點閱:144下載:0
分享至:
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報
  • 阿魏酸(ferulic acid, 4-hydroxy 3-methoxy cinnamic acid)研究證實具有抗氧化、抗UV、抗癌及降血壓等活性,普遍存在於許多植物中,分子本身para-hydroxyl的結構更增加其抗氧化性,阿魏酸為有效之自由基清除劑,可防止自由基對神經元細胞的攻擊而不損害細胞本身。OMC是目前研究最為廣泛的UVB防曬劑,也是最不易引起過敏之化學防曬劑。本研究選用阿魏酸及4-甲基肉桂酸(4-methoxycinnamic acid)為基質,以Novozym 435酵素催化酯化反應分別進行合成乙基阿魏酸酯(ethyl ferulate)及辛基肉桂酸酯(octyl methoxycinnamate)。目前文獻有關酵素催化合成上述產物的結果僅達低轉化率且反應時間長,本研究尋求最適化的操作條件而在合理反應時間下達到高轉化率。本研究並探討Novozym 435之重複使用性。


    Ferulic acid (4-hydroxy 3-methoxy cinnamic acid), which occurred naturally in many plants, has been shown to possess photoprotective, antioxidant, antitumor and hypertensive activities. Octyl methoxycinnamate (OMC) is the most widely used non-allergic sunscreen agent in the market. Most literatures reported low conversion and long reaction time in the syntheses of ethyl ferulate and OMC. In his study, Novozym 435 was selected as the catalyst for the syntheses of ethyl ferulate from ferulic acid and ethanol, and OMC from 4-methoxycinnamic acid and 2-ethyl hexanol. Parameters that affect the conversion of these reactions as well as the reusability of Novozym 435 were systematically investigated.

    Subjects catalog Chapter 1 Introduction......1 1-1 Phenolic acid.1 1-2 Antioxidants...2 1-3 Radical scavenging..3 1-4 UV…6 1-5 Enzyme......9 1-6 Ferulic acid and ethyl ferulate…..12 1-7 p-Methoxycinnamic acid and octyl methoxycinnamate...13 1-8 About this research……13 1-9 Chemical structure…14 Chapter 2 Literature survey…16 Chapter 3 Chemicals, equipments and procedures..18 3-1 Chemicals…….18 3-2 Equipments……...19 3-3 Procedures.19 3-4 HPLC analysis…..20 3-5 NMR analysis...22 3-6 Purification 25 3-7 Determine the conversion….25 3-8 Reused lipase activity assay 25 Chapter 4 Results and discussions ...27 4-1 Effect of log P…28 4-2 Effect of temperature………29 4-3 The effects of stirring on enzymatic esterification..31 4-4 Reproducibility problems…...31 4-5 Effect of substrate ratio……..32 4-6 Effect of water……..34 4-7 Effect of alcohol chain length…..35 4-8 Reusabiliy of Novozym 435…….40 4-9 Thermal decarboxylation of ferulic acid…44 Chapter 5 Conclusion…...46 Reference….......47 Figure catalog Fig. 1.1 Resonance stabilization of ferulic acid radical….…4 Fig. 1.2 Penetration of UV radiation into the skii….7 Fig. 1.3 Mechanism of UV protection by absorption….8 Fig. 1.4 Structure of octyl-p-methoxy-trans-cinnamate (E-OMC) and octyl-p-methoxy-cis-cinnamate (Z-OMC)……9 Fig. 1.5 Reaction route having a smaller activation energy is provided by enzyme catalysis 11 Fig. 1.6 Process of enzyme catalyzed reaction…..12 Fig. 3.1 HPLC separation of ferulic acid and ethyl ferulate…21 Fig. 3.2 HPLC separation of p-methoxycinnamic acid and octyl methoxycinnamat…22 Fig. 3.3 1H NMR spectrum of EE.23 Fig. 3.4 1H NMR spectrum of OMC…24 Fig. 4.1 The effect of solvent on lipase-catalyzed synthesis of OMC…..29 Fig. 4.2 The effect of temperature on lipase-catalyzed synthesis of ethyl ferulate..30 Fig. 4.3 The effect of temperature on lipase-catalyzed synthesis of OMC….…31 Fig. 4.4 The effect of molar ratio of substrate on lipase-catalyzed synthesis of ethyl ferulate….33 Fig. 4.5 The effect of water content on the synthesis of EF….35 Fig. 4.6 Effect of water on the lipase-catalyzed synthesis of OMC.35 Fig. 4.7 The effect of n-alcohol chain length on the esterification of p-methoxycinnamic acid...37 Fig. 4.8 The effect of linear and branched chain alcohols on the synthesis of p-methoxycinnamic acid ester derivatives..38 Fig. 4.9 Effect of n-alcohol chain length on the esterification of ferulic acid...39 Fig. 4.10 The effect of linear and branched chain alcohols on the synthesis of ferulic acid ester derivatives.40 Fig. 4.11 Relative activity of Novozym 435 versus temperature.…41 Fig. 4.12 Reusability study of Novozym 435 in the synthesis of OMC…….…42 Fig. 4.13 Mechanism for the conversion of ferulic acid..45 Table catalog Table 2.1 Results of enzymatic cinnamic acid esters syntheses in the literature……………………………………………………………17 Table 3.1 Conversion (%) for OMC between calculation and peak ratio………………………………………………………………...25 Table 3.1 Conversion (%) for EF between calculation and peak ratio.…25

    Reference
    Abella, A., J. Chalas, C. Claise, D. Laurent, L. Vergne, S. Labidalle and A. Lindenbaum (1997), Interest of ethyl esters of polyphenolic acids in prevention of atherosclerosis, Atherosclerosis, 199
    Andreasen, M.F., L.P. Christensen, A.S. Meyer and A. Hansen (1999), Release of hydroxycinnamic and hydroxyl-benzoic acids in rye by commercial plant cell wall degrading enzyme preparations. J Sci Food Agr, 79, 411-413
    Brzozowski, A.M., U. Derewenda, Z.S. Derewenda, G.G. Godson, D.M. Lawson, J.P. Turkenburg, F. Bjorkling, B. Huge–Jensen, S.A. Patkar and L. Thim (1991), A model for interfacial activation in lipases from the structure of a fungal lipase-inhibitor complex. Nature, 351, 491-494
    Buisman, G.J.H., C.T.W. van Helteren, G.F.H. Kramer, J.W. Veldink, J.T.P. Derksen and F.P. Cuperus (1998), Enzymatic esterifications of functionalized phenols for the synthesis of lipophilic actioxidants, Biotechnol Lett, 131-136
    Chimi, H, J. Cillard, P. Cillard and M. Rahmani (1991), Peroxyl and hydroxyl radical scavenging activity of some natural phenolic antioxidants, J Am Oil Chem Soc, 68, 307-312.
    Chimi, H, A. Sadik, B. Letutour, and M. Rahmani, (1988) Antioxydant activity of tyrosol, hydroxytyrosol, caffeic acid, oleuropein and BHT in olive oil, Revue Francaise Des Corps Gras, 35, 339-344
    Compton, D.L., J.A. Laszlo and M.A. Berhow (2000), Lipase-catalyzed synthesis of ferulate esters, J Am Oil Chem Soc, 513-519
    Compton, D.L., J.A. Laszlo and T.A. Isbell (2004), Cinnamoyl esters of lesquerella and castor oil: novel sunscreen active ingredients, J Am Oil Chem Soc, 945-951
    Denisov, E.T. and I.V. Khudyakov (1987), Mechanisms of action and reactivities of the free-radicals of inhibitors, Chem Rev, 87, 1313
    Dziedzic, S.Z., B.J.F. Hudson (1984), Phenolic acids and related compounds as antioxidants for edible oils, Food Chem, 14, 45-51
    Gandhi, N.N. (1997), Applications of lipase, J Am Oil Chem Soc, 74, 621-633.
    Garcia, T., A. Coteron, M. Martinez and J. Aracil, Kinetic model for the esterification of oleic acid and cetyl alcohol using an immobilized lipase as catalyst, Chem Eng Sci 55 (2000) 1411-1423
    Giuliani, S., C. Piana, L. Setti, A. Hochkoeppler, P.G. Pifferi, G. Williamson and C.B. Faulds (2001), Synthesis of pentylferulate by a feruloyl esterase from Aspergillus niger using water-in-oil microemulsions, Biotechnol Lett, 23, 325–330.
    Graf, E. (1992), Antioxidant potential of ferulic acid, Free Radical Bio Med, 13, 435-448
    Guyot, B., B. Bosquette, M. Pina and J. Graille (1997), Esterification of phenolic acids from green coffee with an immobilized lipase from Candida antarctica in solvent-free medium, Biotechnol Lett, 529-532
    Hansch, C., A. Leo, and D. Hoekman (1995), Exploring QSAR. Hydrophobic, Electronic and Steric Constants. ACS Professional Reference Book, Washington, D.C.
    Hatzakis, N.S., D. Daphnomili, I. Smonou (2003), Ferulic acid esterase from Humicola insolens catalyzes enantioselective transesterifcation of secondary alcohols, J Mol Catal B: Enzym, 21, 309-311
    Howard, J.A. (1972), Electron-spin resonance-spectrum of tert-butylperoxy radical, Can J Chem, 50, 1981.
    Hsiao, G., M.Y. Shen, W.C. Chang, Y.W. Cheng, S.L. Pan, Y.H. Kuo, T.F. Chen and J.R. Sheu (2003), A novel antioxidant, octyl caffeate, suppression of LPS/IFN-r-induced inducible nitric oxide synthase gene expression in rat aortic smooth muscle cells, Biochem Pharmacol, 1383-1392
    Ishigami, T., K. Nakazato, M. Uehara and T. Endo (1979), Marked ependence of multiplicity in direst Z,E photoisomerization of a series of methyl cinnamates on their para-substitutents, Tetrahedron Lett, 20, 863–866.
    Jayaprakasha, G.K. and L.J. Rao (2000), Phenolic constituents from lichen Parmotrema stuppeum (Nyl.) Hale and their antioxidant activity. , 55, 1018–1022.
    Jimenez, M.M., J. Pelletier, M.F. Bobin and M.C. Martini (2004), Influence of encapsulation on the in vitro percutaneous absorption of octyl methoxycinnamate, Int J Pharm, 45-55
    Klibanov A.M. (2001), Improving enzymes by using them in organic solvents, Nature, 409, 241-246
    Kikuzaki, H., M. Hisamoto, K. Hirose, K. Akiyama and H. Taniguchi (2002), Antioxidant properties of ferulic acid and its related compounds, Agri Food Chem, 2161-2168
    Koseki, T., Y. Ito, S. Furuse, K. Itoand K. Iwano, Conversion of ferulic acid into 4-vinylguaiacol, vanillin and vanillic acid in model solutions of Shochu, J Ferment Bioeng, 82 (1996) 42-50
    Kovac, A., H. Scheib, J. Pleiss, R.D. Schmid and F. Paltauf, Molecular basis of lipase stereoselectivity, Eur J Lipid Sci Technol 102 (2000) 61-77
    Langner, R. and H. Surburg (1997), Naturally Occurring Isoamyl p-Methoxycinnamate: An effective organic-chemical UV absorber can be detected in a plant, Cosmetics and Toiletries, 74, 112.
    Lee, Y.J., H.C. Kuo, C.Y. Chu, C.J. Wang, W.C. Lin and T.H. Tseng (2003), Involvement of tumor suppressor protein p53 and p38 MAPK in caffeic acid phenethyl ester-induced apoptosis of C6 glioma cells, Biochem Pharmacol, 2281-2289
    Liu, L., W.R. Hudgins, S. Shack, M.Q. Yin and D. Samid (1995), Cinnamic acid - a natural product with potential use in cancer intervention, Int J Cancer, 62, 345-350
    Lue, B.M., S. Karboune, F.K. Yeboah and S. Kermasha (2005), Lipase catalyzed esterification of cinnamic acid and oleyl alcohol in organic solvent media, J Chem Technol Biot, 80, 462-468
    Mahapatro, A., B. Kalra, A. Kumar and R.A. Gross (2003), Lipase catalyzed polycondensations: effect of substrates and solvent on chain formation, dispersity, and end-group structure, Biomacromolecules, 4, 544-551
    Mahoney, L.R. (1969), 98Antioxidants, Angew Chem Int Edit, 8, 547
    Margovich, M.J., A. Jimenez, K. Gray and G.J. Kellof (2000), Efficacy of potential chemopreventive agents on rat colon aberrant crypt formation and progression, Carcinogenesis, 1149
    MartiMestres, G., C. Fernandez, N. Parsotam, F. Nielloud, J.P. Mestres and H. Maillols (1997), Stability of UV filters in different vehicles: Solvents and emulsions, Drug Dev Ind Pharm, 23, 647–655.
    Marty, A., W. Chulalaksananukul, R.M. Willemot and J.S. Condoret (1992), Kinetics of lipase-catalyzed esterification in supercritical CO2. Biotechnol Bioeng 39, 273–280
    Mori, H., K. Kawabata, N. Yoshimi, T. Tanaka, T. Murakami, T. Okada and H. Muray (1999), Chemopreventive effects of ferulic acid on oral and rice germ on large bowel carcinogenesis, Anticancer Res, 3775
    Narayan, V. S. and A. M. Klibanov (1993), Are water immiscibility and apolarity of the solvent relevant to enzyme efficiency? Biotechnol Bioeng, 41, 390-393
    Novozym 435 product sheet, Enzyme Business, Novo Nordisk company (1999).
    Otterstedt, J.E. (1973), Photostability and molecular structure, J Phys Chem, 58, 5716- 5725
    Pattanaargson, S., T. Munhapol, P. Hirunsupachot, P. Luangthongaram (2004), Photoisomerization of octyl methoxycinnamate, J Photoch Photobio A, 161, 269–274
    Pokorny, J. (1988) in Autoxidation of Unsaturated Lipids (Chan, H., ed.), Academic Press, London, pp. 141-206
    Porter, N.A. (1986), Mechanisms for the autoxidation of polyunsaturated lipids, Acc Chem Res, 19, 262-268
    Priya, K. and A. Chadha (2003), Synthesis of hydrocinnamic esters by Pseudomonas cepacia lipase, Enzyme Microb Tech, 485-490
    Pyysalo, T., H. Torkkeli and E. Honkanen (1997), The thermal decarboxyboxylation of some substituted cinnamic acids. Lebens-Wissensch Technol, 10, 438-441
    Ramamurthi, S. and A.R. McCurdy (1994), Lipase-catalyzed esterification of oleic acid and methanol in hexane. A kinetic study. J Am Oil Chem Soc, 71, 927-930.
    Reyes, H. R. and C.G.J. Hill (1994), Kinetic modeling of interesterification reactions catalyzed by immobilized lipase. Biotechnol Bioeng, 43, 171-182
    Rybka, K., J. Sitarski and K. Raczynska-Bojanowska (1993), Ferulic acid in rye and wheat grain and grain dietary fiber. Cereal Chem, 70, 55–59.
    Saija, A., A. Tomaino, D. Tromdetta, A. De Pasquale, N. Uccella, T. Barbuzzi, D. Paolino and F. Bonita (2000), In vitro and in vivo evaluation of caffeic and ferulic acids as topical photoprotective agents, Int J Pharm, 39
    Scapagnini, G., D.A. Butterfield, C. Colombrita, R. Sultana, A. Pascale and V. Calabrese (2004), Ethyl ferulate, a lipophilic polyphenol, induces HO-1 and protects rat neurons against oxidative stress, Antioxid Redox Signal, 811-818
    Scharfetter-Kochanek, K., J. Wenk, B. Brenneisen, R. Blaudschun and M. Wlaschek, Molecular role of reactive oxygen species in photoaging and tumor progression. In: P. Altmeyer et al. (Eds.). Skin Cancer and Radiation. Springer, Berlin-Heidelberg (1997) pp. 115–127
    Schmid, R.D., and R. Verger (1998), Lipases: interfacial enzymes with attractive applications, Angew. Chem. Int. Ed., 37, 1608-1633.
    Schudel, P., Grut, R. Schwarzenbach, Winterthur, Hans U. Gonzenbach, Geneva (1987), Process for treating 2-ethylhexyl p-methoxycinnamate in the presence of a phenol, U.S. Patent.
    Schuler, P., and B.J.F. Hudson, Food antioxidants, Elsevier, London, 1990 113-127
    Serkedjieva J., N. Manolova and V. Bankova (1992), Anti-influenza virus effect of some propolis constituents and their analogues (esters of substituted cinnamic acids), J Natl Prod, 55, 294-302.
    Shahidi, F., Natural antioxidants (1996), 102
    Stamatis, H., V. Sereti and F.N. Kolisis (2001), Enzymatic synthesis of hydrophilic and hydrophobic derivatives of natural phenolic acids in organic media, J Mol Catal B: Enzym, 323-328
    Stamatis, H., V. Sereti, and F.N. Kolisis (1999), Studies on the enzymatic synthesis of lipophilic derivatives of natural antioxidants, J Am Oil Chem Soc, 1505-1510
    Streilein, J.W., J.R. Taylor, V. Vincek, I. Kurimoto, T. Shimizu, C. Tie and C. Coulomb (1994), Immune surveillance and sunlight-induced skin cancer. Immunol Today, 15, 174–179
    Soumanou, M.M. and U.T. Bornscheuer (2003), Improvement in lipase-catalyzed synthesis of fatty acid methyl esters from sunflower oil, Enzyme and Microbial Technology, 33, 97–103
    Susuki, A., D. Kagawa, A. Futji, R. Ochiai, I. Tokimitsu and I. Saito (2002), Short- and long-term effects of ferulic acid on blood pressure in spontaneously hypertensive rats, Am J Hypertens, 351
    Taniguchi, H., Wakayama-ken, E. Nomura, Wakayama, T. Tsuno and S. Minami, Ferulic acid ester antioxidant/UV absorbent, United States Patent, 1999
    Tarras-Wahlberg, N., G. Stenhagen, O. Larko, A. Rosen, A.M. Wennberg and O. Wennerstrom (1999), Changes in ultraviolet absorption of sunscreens after ultraviolet irradiation, J Invest Dermatol, 113, 547– 553.
    Timasheff, S. N. (1993), The control of protein stability and associations by weak interactions with water: how do solvents affect these processes? Annu Rev Bioph Biom Struct, 22, 67-97
    Topakas, E., H. Stamatis, P. Biely, D. Kekos, B.J. Macris, P. Christakopoulos (2003), Purification and characterization of a feruloyl esterase from Fusarium oxysporum catalyzing esterification of phenolic acids in ternary water-organic solvent mixtures, J Biotechnol, 102, 33-44.
    Volkin, D. B., A. Staubli, R. Langer and A.M. Klibanov (1991), Enzyme thermoinactivation in anhydrous organic solvent. Biotechnol Bioeng 37, 843-853
    Watanabe, Y., Y. Shimada, A. Sugihara, and Y. Tominaga (1999), Stepwise ethanolysis tuna oil using immobilized Candida antarctica lipase, Journal of Bioscience and Bioengineering, 6, 622-626.
    Yadav, G.D. and A.H. Trivedi, Kinetic modeling of immobilized-lipase catalyzed transesterification of n-octanol with vinyl acetate in non-aqueous media, Enzyme Microb Technol 32 (2003) 783–789
    Zaidi, A., J.L. Gainer, G. Carta, A. Mrani, T. Kadiri, Y. Belarbi and A. Mir (2002), Esterification of fatty acids using nylon-immobilized lipase in n-hexane: kinetic parameters and chain-length effects, J Biotechnol 93, 209-216
    Zaks, A. and A.M. Klibanov (1988), The effect of water on enzyme action in organic media. J Biol Chem 263, 8017-8021
    Zupfer, J.M., K.E. Churchill, D.C. Rasmusson and R.G. Fulcher (1998), Variation in ferulic acid concentration among diverse barley cultivars measured by HPLC and microspectrophotometry. J Agr Food Chem, 46, 1350–1354.

    無法下載圖示
    全文公開日期 本全文未授權公開 (校外網路)

    QR CODE