-The objective of this work was to define the optimal conditions for invertase assay, seeking to determine the ideal parameters for the different isoenzymes of leaf and bark tissues in adult rubber trees. Assays of varying pH, sucrose concentration and temperature of the reaction medium were conducted for the two investigated isoenzymes. The results pointed out the existence of two different pH related isoforms for the two analyzed tissues, with an isoenzyme being more active at pH 5,5 and the other at neutral/alkaline pH. Leaf blade isoenzymes presented similar values for substrate concentration, whereas the bark isoenzyme presented maximum values below those previously reported. The assays at different temperatures presented similar values for leaf isoenzymes, though they have differed significantly among the obtained values.Keywords: invertase assay, isoenzyme, rubber trees. OTIMIZAÇÃO DAS CONDIÇÕESS DO ENSAIO DA INVERTASE EMSERINGUEIRA (Hevea brasiliensis Muell. Arg.) 1 RESUMO -O objetivo deste trabalho foi definir as condições ótimas para a realização do ensaio enzimático da invertase, procurando-se determinar os parâmetros ideais para as diferentes isoenzimas de tecidos foliares
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