Abstract
Escherichia coli W3110 was previously engineered to produce xylitol from a mixture of glucose plus xylose by expressing xylose reductase (CbXR) and deleting xylulokinase (ΔxylB), combined with either plasmid-based expression of a xylose transporter (XylE or XylFGH) (Khankal et al., J Biotechnol, 2008) or replacing the native crp gene with a mutant (crp*) that alleviates glucose repression of xylose transport (Cirino et al., Biotechnol Bioeng 95:1167–1176, 2006). In this study, E. coli K-12 strains W3110 and MG1655 and wild-type E. coli B were compared as platforms for xylitol production from glucose-xylose mixtures using these same strategies. The engineered strains were compared in fed-batch fermentations and as non-growing resting cells. Expression of CRP* in the E. coli B strains tested was unable to enhance xylose uptake in the presence of glucose. Xylitol production was similar for the (crp*, ΔxylB)-derivatives of W3110 and MG1655 expressing CbXR (average specific productivities of 0.43 g xylitol g cdw−1 h−1 in fed-batch fermentation). In contrast, results varied substantially between different ΔxylB-derivative strains co-expressing either XylE or XylFGH. The differences in genetic background between these host strains can therefore profoundly influence metabolic engineering strategies.
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Acknowledgments
We acknowledge W. Boos (University of Konstanz) for providing strain ET25. This research was made possible by a grant from the NSF (BES0519516) and support from the Penn State Associate Vice President for Research.
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Reza Khankal and Francesca Luziatelli contributed equally to the paper.
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Khankal, R., Luziatelli, F., Chin, J.W. et al. Comparison between Escherichia coli K-12 strains W3110 and MG1655 and wild-type E. coli B as platforms for xylitol production. Biotechnol Lett 30, 1645–1653 (2008). https://doi.org/10.1007/s10529-008-9720-7
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DOI: https://doi.org/10.1007/s10529-008-9720-7