Publication: Clonación, expresión, caracterización y modelado de
esterasas putativas de Corynebacterium glutamicum ATTC
13032
Authors
Navarro-González, Inmaculada
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Publisher
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DOI
http://dx.doi.org/10.6018/analesbio.0.34.12
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info:eu-repo/semantics/article
Description
Abstract
El objetivo de este trabajo fue clonar, expresar, purificar y caracterizar
algunos genes de Corynebacterium glutamicum ATCC13032,
identificados como enzimas de la superfamilia de las esterasas/lipasas.
De todos los genes clonados, solo se pudo estudiar la proteína
codificada por el gen Cg2528. Se purificó y caracterizó la
proteína. Su máxima actividad la presentó 40 ºC y a pH 8. Presentó
una gran estabilidad en presencia de disolventes orgánicos como
el DMSO y el metanol. Con respecto al análisis cinético, presentó
una mayor actividad sobre los p-nitrofenil ésteres y los naftilos
que sobre el S-methiltiobutanoato. Se dilucidó la triada catalítica
alineando la secuencia de aminoácidos. La estructura tridimensional
se predijo usando la esterasa (2OGS) de Geobacillus stearothermophilus
como molde.
Abstract Cloning, expression, characterization and modeling of putatives esterases from Corynebacterium glutamicum ATCC 13032 The aim of this work was to clone, express, purifiy and characterize some genes from Corynebacterium glutamicum ATCC13032, identified as enzymes of the superfamily of esterases/lipases. Among all cloned genes, only the protein encoded by the gene Cg-2528 could be studied. The protein was purified and characterized. The optimum activity was observed at 40ºC and pH 8. The enzyme showed a high activity in presence of organic solvents, such as DMSO or methanol. P-nitrophenyl ester and naphthyl ester derivatives were found to be better substrates than S-methyl tiobutanoate. The catalytic triad was predicted by amino acid sequence alignment and the structure modeling was performed using esterase (2OGS) form Geobacillus stearothermophilus as template.
Abstract Cloning, expression, characterization and modeling of putatives esterases from Corynebacterium glutamicum ATCC 13032 The aim of this work was to clone, express, purifiy and characterize some genes from Corynebacterium glutamicum ATCC13032, identified as enzymes of the superfamily of esterases/lipases. Among all cloned genes, only the protein encoded by the gene Cg-2528 could be studied. The protein was purified and characterized. The optimum activity was observed at 40ºC and pH 8. The enzyme showed a high activity in presence of organic solvents, such as DMSO or methanol. P-nitrophenyl ester and naphthyl ester derivatives were found to be better substrates than S-methyl tiobutanoate. The catalytic triad was predicted by amino acid sequence alignment and the structure modeling was performed using esterase (2OGS) form Geobacillus stearothermophilus as template.
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