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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 glucose and acetate 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 (50Glu), 10 g l−1 acetate (10Ace) or both (50Glu–10Ace); or 40 g l−1 glucose and 10 g l−1 acetate (40Glu–10Ace); or 30 g l−1 glucose and 10 g l−1 acetate (30Glu–10Ace). IBA concentration (a), isobutanol concentration (f), consumed glucose (b), consumed acetate (d) and cell growth (e) were monitored during the experiment. Carbon yield of IBA was calculated at 72 h (c) Error bars indicate s.d. (n=3).
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f3: IBA production with glucose and acetate 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 (50Glu), 10 g l−1 acetate (10Ace) or both (50Glu–10Ace); or 40 g l−1 glucose and 10 g l−1 acetate (40Glu–10Ace); or 30 g l−1 glucose and 10 g l−1 acetate (30Glu–10Ace). IBA concentration (a), isobutanol concentration (f), consumed glucose (b), consumed acetate (d) and cell growth (e) were monitored during the experiment. Carbon yield of IBA was calculated at 72 h (c) Error bars indicate s.d. (n=3).

Mentions: On identification that Ack–Pta pathway was the best acetate-assimilating pathway in the three tested pathways, it was combined with the IBA synthesis pathway in JCL260 and AL2045 (strains 7 and 8, respectively, Table 1). To identify a good condition for IBA production varying concentrations of glucose and acetate were investigated. The strains were grown in M9P (modified M9 minimal media see Methods) with glucose, acetate or both for 3 days and IBA production was monitored (Fig. 3, Supplementary Fig. 2). The carbon yield of IBA (CYIBA) is defined with the following formula:


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

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

IBA production with glucose and acetate 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 (50Glu), 10 g l−1 acetate (10Ace) or both (50Glu–10Ace); or 40 g l−1 glucose and 10 g l−1 acetate (40Glu–10Ace); or 30 g l−1 glucose and 10 g l−1 acetate (30Glu–10Ace). IBA concentration (a), isobutanol concentration (f), consumed glucose (b), consumed acetate (d) and cell growth (e) were monitored during the experiment. Carbon yield of IBA was calculated at 72 h (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

f3: IBA production with glucose and acetate 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 (50Glu), 10 g l−1 acetate (10Ace) or both (50Glu–10Ace); or 40 g l−1 glucose and 10 g l−1 acetate (40Glu–10Ace); or 30 g l−1 glucose and 10 g l−1 acetate (30Glu–10Ace). IBA concentration (a), isobutanol concentration (f), consumed glucose (b), consumed acetate (d) and cell growth (e) were monitored during the experiment. Carbon yield of IBA was calculated at 72 h (c) Error bars indicate s.d. (n=3).
Mentions: On identification that Ack–Pta pathway was the best acetate-assimilating pathway in the three tested pathways, it was combined with the IBA synthesis pathway in JCL260 and AL2045 (strains 7 and 8, respectively, Table 1). To identify a good condition for IBA production varying concentrations of glucose and acetate were investigated. The strains were grown in M9P (modified M9 minimal media see Methods) with glucose, acetate or both for 3 days and IBA production was monitored (Fig. 3, Supplementary Fig. 2). The carbon yield of IBA (CYIBA) is defined with the following formula:

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