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Two-dimensional isobutyl acetate production pathways to improve carbon yield.

Tashiro Y, Desai SH, Atsumi S - Nat Commun (2015)

Bottom Line: To avoid these problems, we describe here the construction of a metabolic pathway that simultaneously utilizes glucose and acetate.We demonstrate the utility of this approach for isobutyl acetate (IBA) production, wherein IBA production with glucose and acetate achieves a higher carbon yield than with either sole carbon source.These results highlight the potential for this multiple carbon source approach to improve the TMCY and balance redox in biosynthetic pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, California 95616, USA.

ABSTRACT
For an economically competitive biological process, achieving high carbon yield of a target chemical is crucial. In biochemical production, pyruvate and acetyl-CoA are primary building blocks. When sugar is used as the sole biosynthetic substrate, acetyl-CoA is commonly generated by pyruvate decarboxylation. However, pyruvate decarboxylation during acetyl-CoA formation limits the theoretical maximum carbon yield (TMCY) by releasing carbon, and in some cases also leads to redox imbalance. To avoid these problems, we describe here the construction of a metabolic pathway that simultaneously utilizes glucose and acetate. Acetate is utilized to produce acetyl-CoA without carbon loss or redox imbalance. We demonstrate the utility of this approach for isobutyl acetate (IBA) production, wherein IBA production with glucose and acetate achieves a higher carbon yield than with either sole carbon source. These results highlight the potential for this multiple carbon source approach to improve the TMCY and balance redox in biosynthetic pathways.

No MeSH data available.


Related in: MedlinePlus

IBA production with acetate-feeding in strain 7.Strain 7 (JCL260 harbouring IBA production and acetate-assimilating pathways, Table 1) was grown in M9P media with 50 g l−1 glucose and 10 g l−1 acetate, where acetate was fed daily. IBA concentration (a), isobutanol concentration (f), consumed glucose (b), consumed acetate (d) and cell density (e) were monitored during the experiment. Carbon yield of IBA was calculated during the entire experiment (c). Error bars indicate s.d. (n=3).
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f5: IBA production with acetate-feeding in strain 7.Strain 7 (JCL260 harbouring IBA production and acetate-assimilating pathways, Table 1) was grown in M9P media with 50 g l−1 glucose and 10 g l−1 acetate, where acetate was fed daily. IBA concentration (a), isobutanol concentration (f), consumed glucose (b), consumed acetate (d) and cell density (e) were monitored during the experiment. Carbon yield of IBA was calculated during the entire experiment (c). Error bars indicate s.d. (n=3).

Mentions: Strain 7 grown with 50 g l−1 glucose and 10 g l−1 acetate allowed for the best IBA titre in the tested conditions (Fig. 3). To convert all 50 g l−1 glucose (0.27 M) to IBA, 10 g l−1 acetate (0.17 M) is not molecularly sufficient. (Fig. 3a,b,d). Additional acetate would be required for IBA production, however, increasing acetate concentration in the media is unfavourable for E. coli growth (Supplementary Fig. 1). Therefore, in the following experiments, acetate concentration was maintained at 10 g l−1 by feeding acetate every 24 h and IBA production was monitored for 5 days (Fig. 5, Supplementary Figs 4 and 5).


Two-dimensional isobutyl acetate production pathways to improve carbon yield.

Tashiro Y, Desai SH, Atsumi S - Nat Commun (2015)

IBA production with acetate-feeding in strain 7.Strain 7 (JCL260 harbouring IBA production and acetate-assimilating pathways, Table 1) was grown in M9P media with 50 g l−1 glucose and 10 g l−1 acetate, where acetate was fed daily. IBA concentration (a), isobutanol concentration (f), consumed glucose (b), consumed acetate (d) and cell density (e) were monitored during the experiment. Carbon yield of IBA was calculated during the entire experiment (c). Error bars indicate s.d. (n=3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4491173&req=5

f5: IBA production with acetate-feeding in strain 7.Strain 7 (JCL260 harbouring IBA production and acetate-assimilating pathways, Table 1) was grown in M9P media with 50 g l−1 glucose and 10 g l−1 acetate, where acetate was fed daily. IBA concentration (a), isobutanol concentration (f), consumed glucose (b), consumed acetate (d) and cell density (e) were monitored during the experiment. Carbon yield of IBA was calculated during the entire experiment (c). Error bars indicate s.d. (n=3).
Mentions: Strain 7 grown with 50 g l−1 glucose and 10 g l−1 acetate allowed for the best IBA titre in the tested conditions (Fig. 3). To convert all 50 g l−1 glucose (0.27 M) to IBA, 10 g l−1 acetate (0.17 M) is not molecularly sufficient. (Fig. 3a,b,d). Additional acetate would be required for IBA production, however, increasing acetate concentration in the media is unfavourable for E. coli growth (Supplementary Fig. 1). Therefore, in the following experiments, acetate concentration was maintained at 10 g l−1 by feeding acetate every 24 h and IBA production was monitored for 5 days (Fig. 5, Supplementary Figs 4 and 5).

Bottom Line: To avoid these problems, we describe here the construction of a metabolic pathway that simultaneously utilizes glucose and acetate.We demonstrate the utility of this approach for isobutyl acetate (IBA) production, wherein IBA production with glucose and acetate achieves a higher carbon yield than with either sole carbon source.These results highlight the potential for this multiple carbon source approach to improve the TMCY and balance redox in biosynthetic pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, California 95616, USA.

ABSTRACT
For an economically competitive biological process, achieving high carbon yield of a target chemical is crucial. In biochemical production, pyruvate and acetyl-CoA are primary building blocks. When sugar is used as the sole biosynthetic substrate, acetyl-CoA is commonly generated by pyruvate decarboxylation. However, pyruvate decarboxylation during acetyl-CoA formation limits the theoretical maximum carbon yield (TMCY) by releasing carbon, and in some cases also leads to redox imbalance. To avoid these problems, we describe here the construction of a metabolic pathway that simultaneously utilizes glucose and acetate. Acetate is utilized to produce acetyl-CoA without carbon loss or redox imbalance. We demonstrate the utility of this approach for isobutyl acetate (IBA) production, wherein IBA production with glucose and acetate achieves a higher carbon yield than with either sole carbon source. These results highlight the potential for this multiple carbon source approach to improve the TMCY and balance redox in biosynthetic pathways.

No MeSH data available.


Related in: MedlinePlus