Cloning, nucleotide sequence and primary biochemical characterization of esterase EstA from Ralstonia eutropha CH34 |
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Authors: | Shuishan Song Silke Hein Alexander Steinbüchel |
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Institution: | (1) Institut für Mikrobiologie, Westfälische Wilhelms-Universität Münster, Corrensstraße 3, D-48149 Münster, Germany |
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Abstract: | A genomic library of Ralstonia eutropha CH34 was screened in Escherichia coli S17-1 for esterase activity by using -naphthyl acetate and Fast Blue RR. A 1,711 bp DNA fragment was subcloned from an esterase-positive clone and sequenced. Esterase EstA was encoded by a 825-bp open reading frame and exhibited significant amino acid similarities with the enzymes involved in the meta-cleavage pathway. EstA is composed of 275 amino aicds with a predicted molecular mass of 30785 Da. The optimal pH for EstA was 7.0, and the enzyme retained more than 65% activity when incubated in buffers with pH 3.8–9.2 for 2 h. EstA was active at temperatures up to 80 °C and retained more than 77% activity after exposure to temperatures below 60 °C for 2 h. |
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Keywords: | esterase meta-cleavage pathway Ralstonia eutropha CH34 |
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