IP Library Granted Patent US 12,173,329
Granted Patent B2
US 12,173,329 · App. 18/355,990 · Granted Dec 24, 2024

Acyl-ACP reductase with improved properties

Inventors: Mathew Rude (San Diego, CA); Na Trinh (Burlingame, CA); Andreas Schirmer (San Diego, CA); Jacob Gano (San Diego, CA)
Assignee: Genomatica, Inc.
C12N9/0008C12P7/04C12P7/24C12P7/6409C12Y102/0108
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Quick Facts
Patent No.
US 12,173,329
App. No.
18/355,990
Granted
Dec 24, 2024
Kind
B2
Abstract

The disclosure relates to acyl-ACP reductase (AAR) enzyme variants that result in improved fatty aldehyde and fatty alcohol production when expressed in recombinant host cells. The disclosure further relates to methods of making and using such AAR variants for the production of fatty alcohol compositions having particular characteristics.

Claims (31)

1. A variant acyl-ACP reductase (AAR) polypeptide comprising at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 81, wherein said variant AAR polypeptide comprises one or more mutations at an amino acid position corresponding to position 8, 10, 11, 16, 17, 18, 19, 21, 22, 23, 24, 31, 34, 38, 43, 50, 58, 63, 64, 65, 83, 86, 96, 112, 113, 116, 117, 118, 120, 135, 148, 150, 153, 154, 155, 157, 159, 168, 172, 177, 179, 180, 187, 188, 191, 209, 210, 211, 236, 238, 253, 274, 277, 281, 282, 283, 284, 285, 286, 291, 316, 324, 328, 335, and 337 of SEQ ID NO: 81, and wherein said variant AAR polypeptide catalyzes the conversion of an acyl-ACP to a fatty aldehyde.

2. The variant AAR polypeptide of claim 1 , wherein expression of said variant AAR polypeptide in a recombinant host cell results in a higher titer of a fatty alcohol composition as compared to a titer produced by expression of a wild type AAR polypeptide in a corresponding wild type host cell.

3. The variant AAR polypeptide of claim 2 , wherein said fatty alcohol composition is a C12 or C14 fatty alcohol composition.

4. The variant AAR polypeptide of claim 1 , wherein said variant AAR polypeptide comprises one or more mutations selected from L8A, S10G, L11F, D16L, V17C, V17L, V17N, V17W, S18C, S18F, S18L, S18M, S18T, S18V, S18W, S18Y, R19A, R19H, R19I, R19K, R19M, R19S, R19T, R19V, M21L, G22S, Y23N, D24E, D24P, D24V, D24Y, L31M, L31V, W34F, P38T, D43E, A50Q, R58S, C63A, C63G, C63H, C63R, C63Y, F64V, L65F, N83G, S86G, S96T, A112R, S113K, Q116G, V117L, R118Q, T120S, A135S, T148C, T148E, T148V, G150D, I153P, T154A, Q155C, Q155G, Q155L, T157V, A159V, I168V, C172L, L177H, L179H, G180S, T187V, T188H, T188V, Q191A, L209R, E210Y, A211W, T236C, Q238H, S253N, I274N, Q277V, A281F, A281L, A281V, A281Y, A282T, A282V, E283G, E283K, E283S, M2841, A285V, R286Q, M291V, Q316K, A324T, A324V, A328S, Q335N, L337V, and L337W.

5. The variant AAR polypeptide of claim 4 , wherein said variant AAR polypeptide comprises one or more mutations selected from L8A, D16L, M21L, D24E, D24Y, D24V, D24P, L31V, L31M, W34F, D43E, A50Q, C63A, C63G, C63Y, S86G, A112R, S113K, Q116G, R118Q, T120S, A135S, T148C, T148E, T148V, 1153P, T154A, Q155C, Q155L, T157V, A159V, I168V, C172L, T187V, T188H, T188V, Q191A, L209R, E210Y, A211W, T236C, Q277V, A281L, E283G, E283S, A285V, M291V, A324T, A328S, Q335N, L337V, and L337W.

6. The variant AAR polypeptide of claim 5 , wherein said variant AAR polypeptide comprises an M21L mutation, a C63G mutation, an S113K mutation, aft T154A mutation, an A281L mutation, or an L8A mutation.

7. The variant AAR polypeptide of claim 1 , wherein said variant AAR polypeptide comprises:

(a) an M21L mutation, a C63G mutation, an S113K mutation, a T154A mutation, and an A281L mutation;

(b) an L8A mutation, an M21L mutation, a C63G mutation, an S113K mutation, a T154A mutation, and an A281L mutation;

(c) a D16L mutation, an M21L mutation, a C63G mutation, an S113K mutation, a T154A mutation, and an A281L mutation;

(d) an L8A mutation, a D24V mutation, a C63G mutation, an S113K mutation, a Q155L mutation, and an A281L mutation;

(e) a D24P mutation, an L31M mutation, a C63G mutation, an S113K mutation, a T154A mutation, and an A281L mutation;

(f) an L8A mutation, a D16L mutation, a D24V mutation, a C63G mutation, an S113K mutation, a T154A mutation, and an A281L mutation; or

(g) a D24E mutation, a C63G mutation, an S113K mutation, a T154A mutation, and an A281L mutation.

8. The variant AAR polypeptide of claim 1 , comprising at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 81.

9. A recombinant host cell expressing the variant AAR polypeptide of claim 1 .

10. The recombinant host cell of claim 9 , wherein the recombinant host cell produces a fatty alcohol composition with a titer that is at least 10% greater than the titer of a fatty alcohol composition produced by a host cell expressing a corresponding wild type AAR polypeptide, when cultured in medium containing a carbon source under conditions effective to express the variant AAR polypeptide.

11. The recombinant host cell of claim 10 , wherein the fatty alcohol composition is produced extracellularly.

12. The recombinant host cell of claim 10 , wherein the fatty alcohol composition is produced at a titer of 30 g/L to 250 g/L.

13. A cell culture comprising the recombinant host cell of claim 9 .

14. The cell culture of claim 13 , wherein the cell culture is capable of producing a fatty alcohol composition, and wherein the fatty alcohol composition comprises one or more of a C6, C8, C10, C12, C13, C14, C15, C16, C17, and a C18 fatty alcohol.

15. The cell culture of claim 14 , wherein the fatty alcohol composition comprises:

(a) one or more of a C10:1, C12:1, C14:1, C16:1, and a C18:1 unsaturated fatty alcohol;

(b) an unsaturated fatty alcohol;

(c) a fatty alcohol having a double bond at position 7 in the carbon chain between C7 and C8 from the reduced end of the fatty alcohol; or

(d) a saturated fatty alcohol.

16. The cell culture of claim 14 , wherein the fatty alcohol composition comprises a fatty alcohol having a double bond at position 7 in the carbon chain between C7 and C8 from the reduced end of the fatty alcohol.

17. A method of producing a fatty alcohol composition, the method comprising culturing the recombinant host cell of claim 9 with a carbon source, wherein the fatty alcohol composition is produced at a titer that is greater than the titer of a fatty alcohol composition produced by a host cell expressing a wild-type AAR of SEQ ID NO: 81.

18. The method of claim 17 , further comprising harvesting the fatty alcohol composition.

19. A recombinant host cell expressing the variant AAR polypeptide of claim 4 .

20. A method of producing a fatty alcohol composition, the method comprising culturing the recombinant host cell of claim 19 with a carbon source, wherein the fatty alcohol composition is produced at a titer that is greater than the titer of a fatty alcohol composition produced by a host cell expressing a wild-type AAR of SEQ ID NO:81.

Assignments (5)
SECURITY INTEREST Recorded Feb 10, 2026
From: GENOMATICA, INC.
To: AGAIN BIO APS
Reel/Frame 074708/0001 →
SECURITY INTEREST Recorded Dec 9, 2025
From: GENOMATICA, INC.
To: NOVO HOLDINGS A/S, AS COLLATERAL AGENT
Reel/Frame 073915/0027 →
SECURITY INTEREST Recorded Jun 2, 2025
From: GENOMATICA, INC.
To: OXFORD FINANCE LLC
Reel/Frame 071471/0770 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 6, 2024
From: RUDE, MATHEW; TRINH, NA; GANO, JACOB; SCHIRMER, ANDREAS
To: REG LIFE SCIENCES, LLC
Reel/Frame 068195/0438 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 6, 2024
From: REG LIFE SCIENCES, LLC
To: GENOMATICA, INC.
Reel/Frame 068348/0444 →