Patent · US Active

Single-cell factory for efficiently synthesizing α-aminobutyric acid and construction and application thereof

US10612054B2 · kind B2 · utility

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14Claims
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Key dates

Filing dateNov 17, 2016
Grant dateApr 7, 2020
Priority date
Expiry dateNov 17, 2036

Classification

  • Technology area (CPC C)Chemistry; Metallurgy
  • CPC primaryC12R2001/19
  • WIPO fieldBiotechnology
  • WIPO sectorChemistry

Abstract

The present invention discloses a single-cell factory for efficiently synthesizing α-aminobutyric acid and construction and application thereof, which belong to the technical field of microorganisms. The present invention expresses an L-threonine deaminase gene, an L-amino acid dehydrogenase gene and a dehydrogenase gene for providing cofactor NADH cycle in tandem to construct a recombinant Escherichia coli single-cell factory which is used for efficiently synthesizing α-aminobutyric acid. The expression level of the L-threonine deaminase is optimized and controlled by an RBS sequence, so that the problem of transformation inhibition caused by the rapid accumulation of an intermediate product ketobutyric acid is solved, moreover, the expression level of the dehydrogenase for providing cofactor NADH cycle is optimized and controlled by a promoter and an RBS sequence, consequently, the NADH regeneration rate is increased, and ultimately, yield is increased. Utilizing the single-cell factory to carry out whole-cell transformation can reduce obstacles to substances getting in and out, increase the transformation rate and promote the intracellular cycle of cofactors without requiring ex…

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