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Characterization of catalytic efficiency parameters of brain cholinesterases in tropical fish

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Abstract

Brain cholinesterases from four fish (Arapaima gigas, Colossoma macropomum, Rachycentron canadum and Oreochromis niloticus) were characterized using specific substrates and selective inhibitors. Parameters of catalytic efficiency such as activation energy (AE), k cat and k cat/k m as well as rate enhancements produced by these enzymes were estimated by a method using crude extracts described here. Despite the BChE-like activity, specific substrate kinetic analysis pointed to the existence of only acetylcholinesterase (AChE) in brain of the species studied. Selective inhibition suggests that C. macropomum brain AChE presents atypical activity regarding its behavior in the presence of selective inhibitors. AE data showed that the enzymes increased the rate of reactions up to 1012 in relation to the uncatalyzed reactions. Zymograms showed the presence of AChE isoforms with molecular weights ranging from 202 to 299 kDa. Values of k cat and k cat/k m were similar to those found in the literature.

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Acknowledgments

The authors would like to dedicate this study to Dr. Patrícia Fernandes de Castro (in memoriam) for her invaluable help and to thank Financiadora de Estudos e Projetos (FINEP/RECARCINE), Petróleo do Brasil S/A (PETROBRAS), Secretaria Especial de Aqüicultura e Pesca (SEAP/PR), Conselho Nacional de Pesquisa e Desenvolvimento Científico (CNPq) and Fundação de Apoio à Ciência e Tecnologia do Estado de Pernambuco (FACEPE) for financial support (Grant Numbers: IBPG-1301-2.08/08 FACEPE, IBPG-0523-2.08/11 FACEPE and BFP-0036-2.08/13 FACEPE). Our gratitude also goes to Universidade Federal Rural de Pernambuco and Aqualider, for providing fish juvenile specimens.

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Authors and Affiliations

  1. Laboratório de Enzimologia – LABENZ, Departamento de Bioquímica and Laboratório de Imunopatologia Keizo Asami - LIKA, Universidade Federal de Pernambuco, Campus Universitário, Recife, PE, 50670-901, Brazil

    Caio Rodrigo Dias de Assis, Amanda Guedes Linhares, Vagne Melo Oliveira, Renata Cristina Penha França, Marina Marcuschi, Ranilson Souza Bezerra & Luiz Bezerra Carvalho Jr.

  2. Laboratório de Glicoproteínas, Departamento de Bioquímica, Universidade Federal de Pernambuco, Recife, PE, Brazil

    Elba Verônica Matoso Maciel Carvalho

  3. Unidade Acadêmica de Serra Talhada, Universidade Federal Rural de Pernambuco, Serra Talhada, PE, Brazil

    Juliana Ferreira Santos

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  1. Caio Rodrigo Dias de Assis
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  2. Amanda Guedes Linhares
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Correspondence to Caio Rodrigo Dias de Assis.

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de Assis, C.R.D., Linhares, A.G., Oliveira, V.M. et al. Characterization of catalytic efficiency parameters of brain cholinesterases in tropical fish. Fish Physiol Biochem 40, 1659–1668 (2014). https://doi.org/10.1007/s10695-014-9956-1

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