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The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Patent No.:

US 10612054 B1

PDFFull Text
Date of Patent:
Apr. 07, 2020

Filed:

Nov. 17, 2016

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

Applicant:

Jiangnan University, Wuxi, CN;

Inventors:

Zhiming Rao, Wuxi, CN;

Junping Zhou, Wuxi, CN;

Taowei Yang, Wuxi, CN;

Xian Zhang, Wuxi, CN;

Meijuan Xu, Wuxi, CN;

CaiZhe Zhang, Wuxi, CN;

Yunlong Qi, Wuxi, CN;

Junxian Zheng, Wuxi, CN;

Assignee:

JIANGNAN UNIVERSITY, Wuxi, CN;

Attorney:

IPro, PLLC

Primary Examiner:

Suzanne M Noakes

Assistant Examiner:

Jae W Lee

Int. Cl.
CPC ...
C12P 13/04 (2006.01); C12N 9/06 (2006.01); C12P 7/40 (2006.01); C12N 9/88 (2006.01); C12N 15/73 (2006.01); C12R 1/07 (2006.01); C12R 1/19 (2006.01);
U.S. Cl.
CPC ...
C12P 13/04 (2013.01); C12N 9/0016 (2013.01); C12N 9/88 (2013.01); C12N 15/73 (2013.01); C12P 7/40 (2013.01); C12Y 104/01005 (2013.01); C12Y 403/01019 (2013.01); C12R 1/07 (2013.01); C12R 1/19 (2013.01);
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 recombinantsingle-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 exogenous addition, and the cost is low. Within 20 h, the yield of the recombinantsingle-cell factory in a 5 L fermentation tank is 204 g·L, the space-time yield is 10.2 g·L·h, and a practical effective strategy is provided for industrialized production.

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