INVESTIGADORES
MENZELLA Hugo Gabriel
congresos y reuniones científicas
Título:
Process optimization for enhancing the production of a thermostable steryl glucosidase in Escherichia coli
Autor/es:
SALVADOR PEIRU; FLORENCIA EBERHARDT; ANDRES AGUIRRE; MARIA CASTELLI; SEBASTIAN CERMINATI; GUILLERMO HAILS; HUGO G. MENZELLA
Reunión:
Conferencia; 9th International Conference on Recombinant Protein Production; 2017; 2017
Resumen:
Biodiesels produced from transesterification of vegetable oils have a major problem in quality due to the presence of precipitates, which are mostly composed of steryl glucosides (SGs). We have recently developed an enzymatic method for the efficient removal of SGs from biodiesel, based on the activity of a thermostable β-glycosidase with steryl glucosidase activity (SGase) from Thermococcus litoralis. However, the viability of the implementation of this process at industrial scale relies on the reduction of the production costs of this enzyme. In the present work we describe the development of an Escherichia coli-based manufacturing system, and a high cell density fermentation process to improve SGase production. Strain engineering includes the assessment of different promoters to drive the expression of a codon-optimized gene, and the co-expression of molecular chaperones. A high cell density fermentation process was developed, where improvements in cost and efficiency of SGase production were achieved by the optimization of the fed batch protocol and media composition (carbon source, inducer and post-induction nutrient feeding strategies). A 300-fold increase in the production titers of SGase was achieved, which directly impacts on the costs of the industrial process for treating biodiesel