Engineering and adaptive evolution of Escherichia coli for D-lactate fermentation reveals GatC as a xylose transporter

Utrilla, Jose, Licona-Cassani, Cuauhtemoc, Marcellin, Esteban, Gosset, Guillermo, Nielsen, Lars K. and Martinez, Alfredo (2012) Engineering and adaptive evolution of Escherichia coli for D-lactate fermentation reveals GatC as a xylose transporter. Metabolic Engineering, 14 5: 469-476. doi:10.1016/j.ymben.2012.07.007


Author Utrilla, Jose
Licona-Cassani, Cuauhtemoc
Marcellin, Esteban
Gosset, Guillermo
Nielsen, Lars K.
Martinez, Alfredo
Title Engineering and adaptive evolution of Escherichia coli for D-lactate fermentation reveals GatC as a xylose transporter
Formatted title
Engineering and adaptive evolution of Escherichia coli for D-lactate fermentation reveals GatC as a xylose transporter
Journal name Metabolic Engineering   Check publisher's open access policy
ISSN 1096-7176
1096-7184
Publication date 2012-09
Sub-type Article (original research)
DOI 10.1016/j.ymben.2012.07.007
Volume 14
Issue 5
Start page 469
End page 476
Total pages 8
Place of publication Orlando, FL, United States
Publisher Academic Press
Collection year 2013
Language eng
Formatted abstract
Despite the abundance of xylose in nature, the production of chemicals from C5 sugars remains challenging in metabolic engineering. By deleting xylFGH genes and using adaptive evolution, an efficient E. coli strain capable of producing d-lactate from xylose was engineered. Quantitative proteomics and genome sequencing were used to understand the new phenotype and the metabolic limitations of xylose conversion to d-lactate. Proteomics identified major changes in enzyme concentration in the glycolytic and tricarboxylic acid pathways. Whole genome sequencing of the evolved strain identified a point mutation in the gatC gene, which resulted in a change from serine to leucine at position 184 of the GatC protein. The knockout of gatC in a number of strains and the insertion of the mutation in the non-evolved strain confirmed its activity as a xylose transporter and demonstrated that the mutation is responsible for the high xylose consumption phenotype in the evolved strain. The newly found xylose transporter is a candidate for future strain engineering for converting C5-C6 syrups into valuable chemicals.
Keyword Adaptive evolution
D-Lactate
Genome sequencing
Proteomics
Transport
Xylose
Pyruvate Formate Lyase
Mineral Salts Medium
Ethanol-Production
Phosphotransferase System
Saccharomyces-Cerevisiae
Xylitol Production
Glycolytic Flux
Gene Knockout
Mutant
Phosphorylation
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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