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Manipulating ATP supply improves in situ CO2 recycling by reductive TCA cycle in engineered Escherichia coli.


ABSTRACT: The reductive tricarboxylic acid (rTCA) cycle was reconstructed in Escherichia coli by introducing pGETS118KAFS, where kor (encodes ?-ketoglutarate:ferredoxin oxidoreductase), acl (encodes ATP-dependent citrate lyase), frd (encodes fumarate reductase), and sdh (encodes succinate dehydrogenase) were tandemly conjugated by the ordered gene assembly in Bacillus subtilis (OGAB). E. coli MZLF (E. coli BL21(DE3) ?zwf, ?ldh, ?frd) was employed so that the C-2/C-1 [(ethanol?+?acetate)/(formate?+?CO2)] ratio can be used to investigate the effectiveness of the recombinant rTCA for in situ CO2 recycling. It has been shown that supplying ATP through the energy pump (the EP), where formate donates electron to nitrate to form ATP, elevates the C-2/C-1 ratio from 1.03?±?0.00 to 1.49?±?0.02. Similarly, when ATP production is increased by the introduction of the heterologous ethanol production pathway (pLOI295), the C-2/C-1 ratio further increased to 1.79?±?0.02. In summary, the ATP supply is a rate-limiting step for in situ CO2 recycling by the recombinant rTCA cycle. The decrease in C-1 is significant, but the destination of those recycled C-1 is yet to be determined.

SUBMITTER: Chen CH 

PROVIDER: S-EPMC7031464 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Manipulating ATP supply improves in situ CO<sub>2</sub> recycling by reductive TCA cycle in engineered <i>Escherichia coli</i>.

Chen Ching-Hsun CH   Tseng I-Ting IT   Lo Shou-Chen SC   Yu Zi-Rong ZR   Pang Ju-Jiun JJ   Chen Yu-Hsuan YH   Huang Chieh-Chen CC   Li Si-Yu SY  

3 Biotech 20200219 3


The reductive tricarboxylic acid (rTCA) cycle was reconstructed in <i>Escherichia coli</i> by introducing pGETS118KAFS, where <i>kor</i> (encodes α-ketoglutarate:ferredoxin oxidoreductase), <i>acl</i> (encodes ATP-dependent citrate lyase), <i>frd</i> (encodes fumarate reductase), and <i>sdh</i> (encodes succinate dehydrogenase) were tandemly conjugated by the ordered gene assembly in <i>Bacillus subtilis</i> (OGAB). <i>E. coli</i> MZLF (<i>E. coli</i> BL21(DE3) Δ<i>zwf</i>, Δ<i>ldh</i>, Δ<i>frd<  ...[more]

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