ISBN: 978-83-60043-10-3 - eurobic9

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Eurobic9, 2-6 September, 2008, Wrocław, Poland P61. Peroxidase from Royal Palm Tree Roystonea Regia: Structure and Stability O. Gavel a , L. Zamorano b , L. Sanz c , J. Calvete c , S. Bursakov a , A. Zhadan d , J. Arellano e , M. Roig b , I. Polikarpov f , V. Shnyrov g a Química, REQUIMTE, CQFB/FCT, Universidade Nova de Lisboa, Quinta da Torre, 2829-516, Caparica, Portugal e-mail: olga.gavel@dq.fct.unl.pt b Química Física, Facultad de Química, Universidad de Salamanca, , 37008, Salamanca, Spain c Instituto de Biomedicina, C.S.I.C., Jaime Roig 11, E-46010, Valencia, Spain d Institute for Biological Instrumentation of the Ru, 142290, Pushchino, Moscow region, Russia e Instituto de Recursos Naturales y Agrobiologia, C., 37008, Salamanca , Spain f Instituto de Física de São Carlos, Universidade, SP CEP 13560-970, Brazil, g Bioquímica y Biología Molecular, Universidad de Salamanca, Plaza Doctores de la Reina, 37007, Salamanca, Spain Heme-binding peroxidases (EC 1.11.1.) carry out a variety of biosynthetic and degradative functions using hydrogen peroxide as the electron acceptor. In this work we present data about primary structure (amino acid sequence, carbohydrate composition and places of its binding) together with results of structural stability study of dimeric peroxidase from leafs of royal palm tree Roystonea regia (RPTP). The sequence of peroxidase is quite conserved and attains 55-61 % identity with Oryza sativa (Rice), Zea mays (Maize), Vitis vinifera (Grape) and Spinacia oleracea (Spinach). The structural stability of the peroxidase was studied by differential scanning calorimetry circular dichroism and steady state tryptophan fluorescence. The thermal and chemical folding/unfolding of royal palm peroxidase (RPTP) at pH 7 is reversible processes that involve a highly cooperative transition between folded dimer and unfolded monomers with very high value of free energy stabilization -near of 23 kcal per mol of monomer- at 25 oC. At pH 3 where ion pairs have disappeared due to protonation, thermally induced denaturation of RPTP is irreversible and strongly dependent upon scan rate, suggesting that this process is under kinetic control. Thermodynamic information was extracted in this case by extrapolation kinetic transition parameters to infinite heating rate. Obtained in this manner value of RPTP stability at 25 oC is ca. 8 kcal per mole of monomer lower than at pH 7. With a big reliability this quantity reflects contribution of ion pair interactions in the structure stability of RPTP. From comparison of RPTP stability with other plant peroxidases it was proposed that responsible for unusual high stability of RPTP that enhance its potential use for biotechnological purposes is its dimerization. Acknowledgement: This work was partially supported by the projects SA-06-00-0 ITACYL-Universidad de Salamanca and SA 129A07 (Junta de Castilla y León) and BFU2004-01432 (Ministerio de Educación y Ciencia) Spain. L.S.Z and O. G. are fellowship holders from Junta de Castilla y León, Spain (Ref. EDU/1490/2003) and from Fundação para a Ciência e a Tecnologia, Portugal (Ref. SFRH/BPD/28380/2006), respectively. _____________________________________________________________________ 180
Eurobic9, 2-6 September, 2008, Wrocław, Poland P62. Preparation of Ethyl β-arylamino Crotonates Using Preyssler Heteropolyacids Preyssler”s Acid H14NaP5W30O110 , H3PMo12O40 and H14NaP5W29 MoO110 and its supported as catalysts A. Gharib a, b* , M. Roshani a , M. Jahangir a a b Department of Chemistry , School of Sciences, Islamic Azad University, Mashhad, Iran Agricultural Researches&Services center, Mashhad, Iran e-mail: aligharib5@yahoo.com The catalysts based on heteropolyacids types and related compounds are less corrosive and produce lower amount of wastes than conventional acid catalysts, so they can be used as replacement in ecofriendly processes.The HPA were supported on several carriers in order to use these catalysts in heterogeneous liquid reactions[1], with the advantage of easy product recovery. On the other hand, 4-quinolones are important compounds and valuable synthetic Intermediates for derivatives that have biological activities belonging to various types, e.g.tuberculostatic [2]. Some , 4-quinolones are used as antibacterials 3 , e.g. ciprofloxacine and other 6-fluoroquinolones.The Conrad- Limpach reaction between anilines and a β-ketoester is a general method to synthesize , 4-quinolones [3]. The aromatic amines react with methyl acetoacetate yielding alkyl β-arylaminocrotonates, acetoacetanilides, diphenylureas or , 4-quinolones , depending on the temperature, solvent and molar ratio of the reactants [4]. These compounds can be cyclized , 4-quinolones by heating in diphenyl ether. R 1 R 2 R 3 + NH 2 O O H 3C OC 2H 5 preyssler catal. R 1 R 2 R 3 N H CH 3 O OC 2H 5 Refrences: [1] L. Pizzio, C. Caceres, M. Blanco, Appl. Catal. A: General 167, 283 (1998) . [2] U. Holzgrabe, M. Steinert, Pharmazie. 56, 11 (2001). [3] W. Werner, Tetrahedron 25, 255 (1969). [4] B. Reddy, synth.commun. 29, 2789 (1999). _____________________________________________________________________ 181 R 1 R 2 O R 3 N H CH 3
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ISBN: 978-83-60043-10-3 - eurobic9

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