Increase in furfural tolerance in ethanologenic Escherichia coli LY180 by plasmid-based expression of ThyA

Huabao Zheng, Xuan Wang, Lorraine P. Yomano, Keelnatham T. Shanmugam, Lonnie O. Ingram

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Furfural is an inhibitory side product formed during the depolymerization of hemicellulose by mineral acids. Genomic libraries from three different bacteria (Bacillus subtilis YB886, Escherichia coli NC3, and Zymomonas mobilis CP4) were screened for genes that conferred furfural resistance on plates. Beneficial plasmids containing the thyA gene (coding for thymidylate synthase) were recovered from all three organisms. Expression of this key gene in the de novo pathway for dTMP biosynthesis improved furfural resistance on plates and during fermentation. A similar benefit was observed by supplementation with thymine, thymidine, or the combination of tetrahydrofolate and serine (precursors for 5,10-methylenetetrahydrofolate, the methyl donor for ThyA). Supplementation with deoxyuridine provided a small benefit, and deoxyribose was of no benefit for furfural tolerance. A combination of thymidine and plasmid expression of thyA was no more effective than either alone. Together, these results demonstrate that furfural tolerance is increased by approaches that increase the supply of pyrimidine deoxyribonucleotides. However, ThyA activity was not directly affected by the addition of furfural. Furfural has been previously shown to damage DNA in E. coli and to activate a cellular response to oxidative damage in yeast. The added burden of repairing furfuraldamaged DNA in E. coli would be expected to increase the cellular requirement for dTMP. Increased expression of thyA (E. coli, B. subtilis, or Z. mobilis), supplementation of cultures with thymidine, and supplementation with precursors for 5,10-methylenetetrahydrofolate (methyl donor) are each proposed to increase furfural tolerance by increasing the availability of dTMP for DNA repair.

Original languageEnglish (US)
Pages (from-to)4346-4352
Number of pages7
JournalApplied and Environmental Microbiology
Volume78
Issue number12
DOIs
StatePublished - Jun 2012
Externally publishedYes

Fingerprint

Furaldehyde
furfural
plasmid
plasmids
Plasmids
tolerance
Escherichia coli
DNA
gene
damage
thymidine
Zymomonas mobilis
Zymomonas
Thymidine
repair
fermentation
yeast
genomics
Bacillus subtilis
bacterium

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Food Science
  • Biotechnology
  • Ecology

Cite this

Increase in furfural tolerance in ethanologenic Escherichia coli LY180 by plasmid-based expression of ThyA. / Zheng, Huabao; Wang, Xuan; Yomano, Lorraine P.; Shanmugam, Keelnatham T.; Ingram, Lonnie O.

In: Applied and Environmental Microbiology, Vol. 78, No. 12, 06.2012, p. 4346-4352.

Research output: Contribution to journalArticle

Zheng, Huabao ; Wang, Xuan ; Yomano, Lorraine P. ; Shanmugam, Keelnatham T. ; Ingram, Lonnie O. / Increase in furfural tolerance in ethanologenic Escherichia coli LY180 by plasmid-based expression of ThyA. In: Applied and Environmental Microbiology. 2012 ; Vol. 78, No. 12. pp. 4346-4352.
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