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Clonación, expresión y caracterización de una nueva esterasa derivada de metagenomas de suelos agrícolas colombianos

C. Villamil,P. Portillo,Alvaro Mongui

Published 2016 in Unknown venue

ABSTRACT

The present work had as a main objective to bioprospect metagenomic DNA from microbial communities associated with an agro-ecosystem of national importance. This analysis allowed the production, expression, purification and characterization of a novel enzyme with esterase activity. This enzyme, named here as LipM, was previously identified in metagenomic clones derived from soils dedicated to creole potato ( Solanum pureja ) crops by means of next-generation sequencing and bioinformatics analyses. The coding sequence of the enzyme was cloned into pBADgiii vector and expressed in E. coli as an expression system, allowing to optimize its recombinant production process and its further purification. The enzyme functionally showed a greater affinity for p- nitrophenyl substrates with short-chain fatty acids (<C8). LipM showed full functionality at temperatures between 30 - 37°C and close to physiological pH (between 7.0 and 8.0). This enzyme also exhibited proper stability in the presence of various metal ions, inhibitors and at 0.1% (w/v) of SDS. Its high level of stability in the presence of inhibitors, metal ions and its particular specificity of substrates make it optimal for use in various biotechnology applications. Key words: Metagenomics; esterase enzyme; characterization;  soil; lipase/esterase.

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