Publication Date:
2017-03-03
Description:
The heterologous expression and characterization of a Hormone-Sensitive Lipases (HSL) esterase ( Ba EstB) from the Basidiomycete fungus Bjerkandera adusta is reported for the first time. According to structural analysis, amino acid similarities and conservation of particular motifs, it was established that this enzyme belongs to the (HSL) family. The cDNA sequence consisted of 969 nucleotides, while the gene comprised 1133, including three introns of 57, 50, and 57 nucleotides. Through three-dimensional modeling and phylogenetic analysis, we conclude that Ba EstB is an ortholog of the previously described Rm EstB-HSL from the phylogenetically distant fungus Rhizomucor miehei . The purified Ba EstB was characterized in terms of its specificity for the hydrolysis of different acyl substrates confirming its low lipolytic activity and a noticeable esterase activity. The biochemical characterization of Ba EstB, the DLS analysis and the kinetic parameters determination revealed this enzyme as a true esterase, preferentially found in a dimeric state, displaying activity under alkaline conditions and relative low temperature (pH = 10, 20°C). Our data suggest that Ba EstB is more active on substrates with short acyl chains and bulky aromatic moieties. Phylogenetic data allow us to suggest that a number of fungal hypothetical proteins could belong to the HSL family. In this work, we report for the first time a HSL from a Basidiomycete fungus. The purified Ba EstB was characterized in terms of its specificity for the hydrolysis of different acyl substrates. The biochemical characterization of Ba EstB, this enzyme as a true esterase, preferentially found in a dimeric state, displaying activity under alkaline conditions and relative low temperature. Our data from substrate specificity suggest that for Ba EstB activity, the bulky moiety of the substrate influences the activity of the enzyme. Phylogenetic data allow us to suggest that a number of fungal hypothetical proteins could belong to the HSL family.
Electronic ISSN:
2045-8827
Topics:
Biology
,
Medicine
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