Co-utilization of glucose and xylose increases growth rate without affecting plasmid DNA yield of engineered E. coli
El consumo simultáneo de glucosa y xilosa incrementa la velocidad de crecimiento sin afectar el rendimiento de ADN plasmídico en una cepa de E. coli modificada genéticamente
R.M. Gálvez, T.E. Pablos, J.C. Sigala, A.R. Lara*
Departamento de Procesos y Tecnología, Universidad Autónoma Metropolitana-Cuajimalpa, Av. Vasco de Quiroga 4871, Col. Santa Fe, Del. Cuajimalpa, México, D.F., C.P. 05348, México. *Corresponding author. E-mail: alara@correo.cua.uam.mx Tel. Tel. 26-36-38-00, Fax 26-36-38-00 ext. 3832.
]]> Received December 4, 2013.
Abstract
Escherichia coli strains devoid of PTS system lack of catabolite repression allowing the simultaneous utilization of glucose and other carbohydrates. In this study, the use of xylose and glucose for the production of plasmid DNA (pDNA) was tested using the engineered strain E. coli PTS- GalP+ Δ(recA, deoR). Growth rate and acetate production were similar using glucose or xylose as the carbon source, but pDNA yield (YP/x) was lower in cultures with xylose. However, when both carbon sources were co-utilized, growth rate increased by 39 %, and YP/x recovered, consequently increasing the specific pDNA production rate by 28 %. Therefore, the used of glucose-xylose mixtures is an attractive alternative for pDNA production.
Keywords: mixed sugars, metabolic engineering, pDNA vaccines, PTS.
Resumen
Las cepas de Escherichia coli con el sistema de fosfotransferasa (PTS) inactivo carecen de represión catabólica, lo que permite el consumo simultáneo de glucosa y otros carbohidratos. En el presente estudio, se evaluó el uso de glucosa y xilosa en la producción de ADN plasmídico (ADNp) usando la cepa modificada E. coli PTS- GalP+ Δ(recA, deoR). La velocidad de crecimiento y la producción de acetato fueron similares empleando glucosa o xilosa como única fuente de carbono, pero el rendimiento de ADNp (YP/x) fue menor en cultivos con xilosa. Sin embargo, cuando ambas fuentes de carbono fueron co-utilizadas, la velocidad de crecimiento se incrementó en 359 % y se recuperó YP/x, lo que en consecuencia incrementó en 28 % la velocidad específica de producción de ADNp. Por lo tanto, el uso de mezclas de glucosa-xilosa es una alternativa atractiva para la producción de ADNp.
Palabras clave: mezcla de carbohidratos, ingeniería metabólica, vacunas de ADN, ITS.
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Acknowledgments
This work was supported by CONACyT and PROMEP grants 183911 and 10828, respectively.
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