Genome-wide mapping of furfural tolerance genes in Escherichia coli.
Advances in genomics have improved the ability to map complex genotype-to-phenotype relationships, like those required for engineering chemical tolerance. Here, we have applied the multiSCale Analysis of Library Enrichments (SCALEs; Lynch et al. (2007) Nat. Method.) approach to map, in parallel, the effect of increased dosage for >10(5) different fragments of the Escherichia coli genome onto furfural tolerance (furfural is a key toxin of lignocellulosic hydrolysate). Only 268 of >4,000 E. coli genes (∼ 6%) were enriched after growth selections in the presence of furfural. Several of the enriched genes were cloned and tested individually for their effect on furfural tolerance. Overexpression of thyA, lpcA, or groESL individually increased growth in the presence of furfural. Overexpression of lpcA, but not groESL or thyA, resulted in increased furfural reduction rate, a previously identified mechanism underlying furfural tolerance. We additionally show that plasmid-based expression of functional LpcA or GroESL is required to confer furfural tolerance. This study identifies new furfural tolerant genes, which can be applied in future strain design efforts focused on the production of fuels and chemicals from lignocellulosic hydrolysate.
Chemical and Biological Engineering, University of Colorado Boulder, Boulder, Colorado, United States of America.
Tirzah Y
TY
Glebes
Chemical and Biological Engineering, University of Colorado Boulder, Boulder, Colorado, United States of America.
Nicholas R
NR
Sandoval
Chemical and Biological Engineering, University of Colorado Boulder, Boulder, Colorado, United States of America.
Philippa J
PJ
Reeder
Chemical and Biological Engineering, University of Colorado Boulder, Boulder, Colorado, United States of America.
Katherine D
KD
Schilling
National Bioenergy Center, National Renewable Energy Laboratory, Golden, Colorado, United States of America.
Min
M
Zhang
Chemical and Biological Engineering, University of Colorado Boulder, Boulder, Colorado, United States of America.
Ryan T
RT
Gill
D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics; D005662:Furaldehyde / Q000494:pharmacology*; D056726:Genetic Association Studies; D006360:Heat-Shock Proteins / Q000235:genetics; D009153:Mutagens / Q000494:pharmacology*
D029968:Escherichia coli Proteins; C028715:GroE protein, E coli; D006360:Heat-Shock Proteins; D009153:Mutagens; D005662:Furaldehyde