	US 12,460,213 B2
	Inducible ammonia production from a symbiotic diazotroph, methods of creation and uses thereof
Tim Schnabel, Stanford, CA (US); and Elizabeth Sattely, Stanford, CA (US)
Assigned to The Board of Trustees of the Leland Stanford Junior University Stanford, CA, Stanford, CA (US)
Appl. No. 17/428,927
Filed by The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US)
PCT Filed Feb. 5, 2020, PCT No. PCT/US2020/016856 § 371(c)(1), (2) Date Aug. 5, 2021, PCT Pub. No. WO2020/163509, PCT Pub. Date Aug. 13, 2020.
Claims priority of provisional application 62/801,454, filed on Feb. 5, 2019.
Prior Publication US 2022/0127624 A1, Apr. 28, 2022
Int. Cl. C12N 9/00 (2006.01); A01N 63/20 (2020.01); C12N 9/12 (2006.01); C12N 15/63 (2006.01); C12N 15/74 (2006.01); C12P 3/00 (2006.01)

CPC C12N 15/635 (2013.01) [A01N 63/20 (2020.01); C12N 9/1241 (2013.01); C12N 15/74 (2013.01); C12P 3/00 (2013.01); C12Y 207/07051 (2013.01)]

14 Claims

1. An engineered diazotroph for increased ammonia production comprising:

a diazotrophic organism comprising:

a genome that comprises mutagenesis within a native glutamine synthetase adenylyl transferase gene such that the native glutamine synthetase adenylyl transferase of the diazotrophic organism does not have adenyl removal activity; and

a transgenic genetic circuit that comprises a unidirectional glutamine synthetase adenylyl transferase gene in operable connection with a promoter, wherein the unidirectional glutamine synthetase adenylyl transferase gene is a sequence derived from a glutamine synthetase adenylyl transferase gene from a genome of a bacterium in which the glutamine synthetase adenylyl transferase gene is modified to have nonfunctional adenylyl removal activity but maintain functional adenylyl transferase activity.