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Pyruvate production using engineered Escherichia coli

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논문

Pyruvate production using engineered Escherichia coli

학술지

AMB Express

저자명

Akita, Hironaga; Nakashima, Nobutaka; Hoshino, Tamotsu

초록

<P>Pyruvate plays an essential role in the central carbon metabolism of multiple organisms and is used as a raw material in the chemical, biochemical and pharmaceutical industries. To meet demand, large amounts of pyruvate are produced through fermentation processes. Here we describe a simple and efficient method for producing pyruvate in <I>Escherichia coli</I>. To stop carbon flux from pyruvate to fatty acids, the <I>accBC</I> genes, which encode the enzyme that catalyzes the first step of fatty acid biosynthesis and is essential for vegetative growth, were manipulated within the genome; its native promoter was replaced with the tetracycline (or doxycycline)-regulated promoter and the corresponding transcriptional regulator genes. The resulting strain grew normally in the presence of doxycycline, but showed poor growth upon withdrawal of doxycycline. Using this strain, we developed a high pyruvate producing strain (strain LAFCPCPt-accBC-aceE), in which the tetracycline-regulated promoter was also introduced upstream of <I>aceE</I>, and the <I>ackA</I>-<I>pta</I>, <I>adhE</I>, <I>cra</I>, <I>ldhA</I>, <I>pflB</I> and <I>poxB</I> genes were deleted. After determining the optimal culture conditions for this strain, the final pyruvate concentration reached 26.1&nbsp;g&nbsp;L<SUP>&#x2212;1</SUP> after 72&nbsp;h with a theoretical yield of 55.6&nbsp;%. These levels are high enough to indicate that the developed strain has the potential for application to industrial production of pyruvate.</P>

발행연도

2016

발행기관

Springer Berlin Heidelberg

라이선스

cc-by

ISSN

2191-0855

6

1

페이지

pp.94

주제어

Pyruvate; Escherichia coli; Promoter regulation; Tetracycline; Doxycycline; Fermenter

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1 2023-12-11

논문; 2016-10-08

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