Vitreoscilla hemoglobin enhances ethanol production by Escherichia coli in a variety of growth media

• Published in: Biomass & bioenergy 2012-02, Vol.37, p.1-8
• Main Authors: Arnaldos, Marina, Kunkel, Stephanie A., Wang, Jia, Pagilla, Krishna R., Stark, Benjamin C.
• Format: Article
• Language: English
• Subjects: acid treatment; activated carbon; adenosine triphosphate; Alternative fuels. Production and utilization; Applied sciences; ATP; Biofuel production; Biological and medical sciences; Biomass; Biotechnology; cell growth; Corn fiber hydrolysate; culture media; Energy; Escherichia coli; ethanol; Ethanol production; Ethanologenic E. coli; Exact sciences and technology; fermentation; Fuels; Fundamental and applied biological sciences. Psychology; hemoglobin; Industrial applications and implications. Economical aspects; Miscellaneous; NAD (coenzyme); nutrient content; Vitreoscilla; Vitreoscilla hemoglobin (VHb); xylose
• ISSN: 0961-9534; 1873-2909
• DOI: 10.1016/j.biombioe.2011.12.048

In a previous study, we showed that expression of Vitreoscilla hemoglobin (VHb) in ethanologenic Escherichia. coli strain FBR5 enhanced ethanol production in a limited range of nutrient concentrations. Here enhancement of ethanol production in the VHb expressing strain (TS3) was investigated in a wider variety of growth media. For nutrient (LB) concentrations from 0.25X to 2X (supplemented with 80 g dm−3 xylose), ethanol production was always better in TS3 than in FBR5. The greatest relative advantage of TS3 over FBR5 occurred at 0.25X LB, while the greatest yield for both strains relative to LB concentration occurred at 0.5X LB. In these experiments, the enhancement in ethanol production of TS3 was determined to be due to both an increased ethanol metabolic efficiency and increased cell growth. 2X LB was also supplemented with purified biomass hydrolysate produced by acid treatment and overliming; additional treatment with activated carbon decreased the fermentability of the hydrolysate. Strain TS3 outperformed strain FBR5 in 2X LB at hydrolysate concentrations of 0.2, 0.4 and 0.8 m3 m−3; the greatest yield by TS3 relative to the level of hydrolysate occurred at 0.4 m3 m−3. The concentrations of NAD, NADH, and ATP were measured in both strains to investigate the biochemical basis of the ethanol production enhancement by VHb. The most significant difference found was in the NAD/NADH ratio; this was generally lower for TS3 than for FBR5 earlier in the fermentation, while this pattern was reversed later in the fermentation. ► Vitreoscilla hemoglobin (VHb) enhances bioethanol production in engineered E. coli. ► The VHb enhancement occurs in media with a wide range of LB concentrations. ► The enhancement is due to both enhanced growth and fermentation efficiency. ► The VHb enhancement occurs over a wide range of biomass hydrolysate concentrations. ► The presence of VHb has modest affects on ATP levels and increases the NAD/NADH ratio.