	US 12,173,338 B2
	Genetically engineered bacteria, its construction method and its application in producing NAD⁺ method
Wei Wang, Anhui (CN); Kanglin Wang, Anhui (CN); Minjie Fu, Anhui (CN); Yonghong Jin, Anhui (CN); Feng Tian, Anhui (CN); Heping Jia, Anhui (CN); and Zhihao Hu, Anhui (CN)
Assigned to HEFEI KNATURE BIO-PHARM CO., LTD., Anhui (CN)
Appl. No. 17/256,364
Filed by HEFEI KNATURE BIO-PHARM CO., LTD., Anhui (CN)
PCT Filed Jun. 11, 2020, PCT No. PCT/CN2020/095553 § 371(c)(1), (2) Date Dec. 28, 2020, PCT Pub. No. WO2021/120548, PCT Pub. Date Jun. 24, 2021.
Claims priority of application No. 201911326087.0 (CN), filed on Dec. 20, 2019.
Prior Publication US 2021/0371843 A1, Dec. 2, 2021
Int. Cl. C12N 9/78 (2006.01); C12N 9/00 (2006.01); C12N 9/10 (2006.01); C12N 9/12 (2006.01); C12P 19/36 (2006.01)

CPC C12N 9/78 (2013.01) [C12N 9/1077 (2013.01); C12N 9/1241 (2013.01); C12N 9/93 (2013.01); C12P 19/36 (2013.01); C12Y 204/02012 (2013.01); C12Y 207/07001 (2013.01); C12Y 305/01019 (2013.01); C12Y 603/05001 (2013.01)]

5 Claims

1. A method for producing a NAD⁺ comprising using nicotinamide or/and adenine as a substrate, and using a genetically engineered microorganism strain to produce the NAD⁺, wherein encoding genes of an adenine deaminase in a genome of the genetically engineered microorganism strain are double knocked out, and encoding genes of enzymes in a NAD⁺ synthesis process are incorporated and overexpressed in the genome of the genetically engineered microorganism strain,

wherein the enzymes in the NAD⁺ synthesis process are Nicotinate phosphoribosyl transferase NPT1 and Nicotinic acid mononucleotide adenylyl transferase NMA1, the strain is diploid Saccharomyces cerevisiae, and the encoding genes of the adenine deaminase are two AAH1 alleles in the diploid Saccharomyces cerevisiae.