Construction of a butyrate-producing E. coli strain without the use of heterologous genes

Серегина, Т. А.; Шакулов, Р. С.; Дебабов, В. Г.; Mironov, A. S.

doi:10.1134/s000368381008003x

Cited by 17 publications

(19 citation statements)

References 25 publications

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“…However, clostridia are disadvantageous due to slow growth, a requirement for strictly anaerobic culture conditions, sporulation and relatively limited genetic tools for use in strain improvement and product diversification (Baek et al, 2013;Fischer et al, 2010;Lim et al, 2013;Zhang et al, 2009). To circumvent these problems, E. coli was engineered for fermentative butyrate production (Baek et al, 2013;Lim et al, 2013;Seregina et al, 2010). This was done by deleting E. coli's native fermentation pathways and engineering it to produce enzymes for butyrate formation that catalyse the condensation and reduction of acyl-CoA compounds: AtoB and TesB from E. coli, Hbd and Crt from C. acetobutylicum and Ter from Treponema denticola (Baek et al, 2013;Fischer et al, 2010;Lim et al, 2013;Zhang et al, 2009).…”

Section: Discussionmentioning

confidence: 99%

“…However, the development of clostridia for efficient production of diverse FAs is encumbered by slow growth, a requirement for strictly anaerobic culture conditions, sporulation and relatively limited genetic tools (Baek et al, 2013;Fischer et al, 2010;Lim et al, 2013;Zhang et al, 2009). Consequently, butyrate production was engineered into E. coli (Baek et al, 2013;Lim et al, 2013;Seregina et al, 2010). The reaction sequence used was similar to the butyrate fermentation of Clostridium to the point of butyryl-CoA formation.…”

Section: Introductionmentioning

confidence: 99%

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Fermentative production of short-chain fatty acids in Escherichia coli

et al. 2014

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Escherichia coli was engineered for the production of even-and odd-chain fatty acids (FAs) by fermentation. Co-production of thiolase, hydroxybutyryl-CoA dehydrogenase, crotonase and trans-enoyl-CoA reductase from a synthetic operon allowed the production of butyrate, hexanoate and octanoate. Elimination of native fermentation pathways by genetic deletion (DldhA, DadhE, DackA, Dpta, DfrdC) helped eliminate undesired by-products and increase product yields. Initial butyrate production rates were high (0.7 g l "1 h "1 ) but quickly levelled off and further study suggested this was due to product toxicity and/or acidification of the growth medium. Results also showed that endogenous thioesterases significantly influenced product formation. In particular, deletion of the yciA thioesterase gene substantially increased hexanoate production while decreasing the production of butyrate. E. coli was also engineered to co-produce enzymes for even-chain FA production (described above) together with a coenzyme B 12 -dependent pathway for the production of propionyl-CoA, which allowed the production of odd-chain FAs (pentanoate and heptanoate). The B 12 -dependent pathway used here has the potential to allow the production of odd-chain FAs from a single growth substrate (glucose) in a more energy-efficient manner than the prior methods.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Fermentative production of short-chain fatty acids in Escherichia coli

et al. 2014

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“…Therefore, E. coli , a well‐characterized microorganism, was metabolically engineered in recent studies for heterologous production of butyrate. Past studies has reported relatively low titers of butyrate (0.5–1.3 g/L) (Joung et al, ; Seregina et al, ), while a recent study reported much higher butyrate titer (4.35 g/L) with optimized N‐terminal sequences of heterologous genes and with complete redox cofactor regeneration (Lim et al, ).…”

Section: Bacterial Strains and Plasmids Used In This Studymentioning

confidence: 99%

Butyrate production in engineered Escherichia coli with synthetic scaffolds

Baek

Mazumdar

Lee

et al. 2013

Biotech & Bioengineering

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Butyrate pathway was constructed in recombinant Escherichia coli using the genes from Clostridium acetobutylicum and Treponema denticola. However, the pathway constructed from exogenous enzymes did not efficiently convert carbon flux to butyrate. Three steps of the productivity enhancement were attempted in this study. First, pathway engineering to delete metabolic pathways to by-products successfully improved the butyrate production. Second, synthetic scaffold protein that spatially co-localizes enzymes was introduced to improve the efficiency of the heterologous pathway enzymes, resulting in threefold improvement in butyrate production. Finally, further optimizations of inducer concentrations and pH adjustment were tried. The final titer of butyrate was 4.3 and 7.2 g/L under batch and fed-batch cultivation, respectively. This study demonstrated the importance of synthetic scaffold protein as a useful tool for optimization of heterologous butyrate pathway in E. coli.

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Section: Discussionmentioning

confidence: 99%

“…Consequently, butyrate production was engineered into E. coli (Baek et al, 2013;Lim et al, 2013;Seregina et al, 2010). The reaction sequence used was similar to the butyrate fermentation of Clostridium to the point of butyryl-CoA formation.…”

Section: Introductionmentioning

confidence: 99%

Fermentative production of short-chain fatty acids and methyl ketones in Escherichia coli

Volker

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Dependence on petroleum for fuels is a well-known issue in the United States today. However, petroleum is also used to produce a broad range of chemicals that are used in applications such as plastics, fragrances, surfactants, detergents, food additives, and pharmaceuticals. As the world's supply of petroleum dwindles, we must look to another method of procuring these chemicals. Biorenewable chemical production attempts to fill this void. Shortchain fatty acids and ketones are desirable precursors to many of these industrially relevant chemicals. Short-chain fatty acids are precursors to alpha-olefins, which are used as lubricants and surfactants in a variety of industries, including the automotive industry. They can also be used as precursors to fatty alcohols, which have potential applications as biodiesel. Methyl ketones are also a class of chemicals with many industrial applications. Butanone is a common industrial solvent, while 4-hydroxybutanone is used in pesticides, terpenoids, and most importantly, is an intermediate in the production of doxorubicin, an anticancer agent. Here we report the biorenewable production of short-chain fatty acids and methyl ketones from fermentation in Escherichia coli. A series of synthetic constructs were made to produce the desired metabolites utilizing glucose as the feedstock. Butyrate was produced at 9.670 g/L, hexanoate at 1.963 g/L, and octanoate at 0.216 g/L. In addition, 0.201 g/L of valerate was produced. Heptanoate production by fermentation in E. coli was reported for the first time, reaching a titer of 0.008 g/L. 4-hydroxy-2-butanone was produced by fermentation at a titer of 2.5 mM. To our knowledge this is the first report of production of 4-hydroxy-2-butanone by microbial fermentation.

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Construction of a butyrate-producing E. coli strain without the use of heterologous genes

Cited by 17 publications

References 25 publications

Fermentative production of short-chain fatty acids in Escherichia coli

Fermentative production of short-chain fatty acids in Escherichia coli

Butyrate production in engineered Escherichia coli with synthetic scaffolds

Fermentative production of short-chain fatty acids and methyl ketones in Escherichia coli

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