Granted Patent B2

US 12,486,212 · App. 18/752,282 · Granted Dec 2, 2025

Inventors: Jeung Yeop Shim (Tampa, FL); Ki Soo Park (Yongin, KR); Ashok Somasundar (Suwon, KR); Sung Hoon Park (Ulsan, KR)

Assignee: NOROO IC CO., LTD.

C07C51/48B01D11/04C07C51/377C07C59/01C12N15/78B01D5/0027B01D17/0214C07C57/04C12N2830/002

Abstract

Claims (16)

1 . A nucleic acid, comprising a first and a second promoter, wherein the first promoter is an inducible promoter and is inducible by a small molecule; the first and second promoters are operably linked to a first gene; and the first gene encodes a protein involved in the synthesis of 3-hydroxypropionic acid (3-HP), a salt of 3-HP, or coenzyme B12 wherein the nucleic acid comprises a sequence selected from the group consisting of SEQ ID NO:22-28 and 64 (UTR 0-6).

2 . The nucleic acid of claim 1 , wherein the first gene encodes a protein involved in the synthesis of 3-hydroxypropionic acid (3-HP) and is selected from the group consisting of a glycerol dehydratase, a glycerol dehydratase reactivase, and an aldehyde dehydrogenase,

wherein:

the glycerol dehydratase is selected from the group consisting of dhaB1, dhaB2, dhaB3;

the glycerol dehydratase reactivase is selected from the group consisting of gdrA and gdrB; and

the aldehyde dehydrogenase is selected from the group consisting of kgsA.

3 . The nucleic acid of claim 1 , wherein the first gene comprises a dhaB1 gene, a dhaB2 gene, a dhaB3 gene, a gdrA gene, and a gdrB gene, and the sequence comprises a sequence that is at least 95% identical to a SEQ ID selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 5 and SEQ ID NO: 6.

4 . The nucleic acid of claim 1 , wherein the small molecule acid or alcohol is selected from the group consisting of L-lactic acid (LAC), acetic acid (AcOH), propionic acid (PA), 3-hydroxypropionic acid (3-HP), 3-hydroxybutyrate (3-HB), 1,3-propanediol (1,3-PDO), 2,3-butanediol (2,3-BDO), L-valine (L-val), and 3-hydroxyisobutyrate (3-HIB).

5 . The nucleic acid of claim 1 , wherein the second promoter is derived from a P mmsA promoter, a P hbdH-1 promoter, a P hbdH-4 promoter, a P hpdH promoter, or a P zwf promoter, and the promoter comprises a sequence that it at least 95% identical to a SEQ ID selected from the group consisting of: SEQ ID NO: 7; SEQ ID NO: 8; SEQ ID NO: 9; SEQ ID NO: 10; SEQ ID NO: 11; SEQ ID NO: 12; SEQ ID NO: 13; SEQ ID NO: 14; SEQ ID NO: 15; SEQ ID NO: 16; SEQ ID NO: 17; SEQ ID NO: 18; SEQ ID NO: 19; SEQ ID NO: 52-63; and SEQ ID NO: 65-72.

6 . The nucleic acid of claim 1 , wherein the nucleic acid further comprises a gene encoding a transcriptional regulator that regulates expression of the first gene, wherein the transcriptional regulator binds to the first or second promoter.

7 . The nucleic acid of claim 6 , wherein the transcriptional regulator is a LysR-type transcriptional regulator (LTTR), a MmsR regulator or a HpdR regulator.

8 . The nucleic acid of claim 6 , wherein the transcriptional regulator binds to the first promoter and has enhanced binding to the first promoter in the presence of the small molecule.

9 . The nucleic acid of claim 6 , wherein the transcriptional regulator is self-regulating.

10 . The nucleic acid of claim 1 , wherein the first gene is fused at its 5′-terminus to a sequence encoding up to 20 (5, 10, 15, or 20) amino acids that are derived from the 5′terminus of a second gene encoding a native Pseudomonas denitrificans protein, thereby creating a fusion gene, and the second promoter is derived from the native promoter for the second gene, and the fusion gene comprises a sequence encoding at least 5, 10, 15, or 20 amino acids in the N-terminus of a native protein.

11 . The nucleic acid of claim 1 , wherein the second promoter comprises the P mmsA promoter and the fusion gene comprises a sequence encoding five or more amino acids from the N-terminus of a native MmsA gene, and the fusion gene is operably linked to the 3′ end of the P mmsA promoter, and wherein the nucleic acid comprises a sequence selected from the group consisting of SEQ ID NOs: 32-35.

12 . The nucleic acid of claim 11 , wherein the mRNA of the fusion gene has a higher stability and increases the gene translation when compared to the first gene alone.

Assignments (3)

Continuity (6)

References Cited (128)