IP Library › Granted Patent US 12,460,239
Granted Patent B2
US 12,460,239 · App. 18/767,621 · Granted Nov 4, 2025

UDP-glycosyltransferase variants and uses thereof

Inventors: Sean Lund (Emeryville, CA); Gale Wichmann (Emeryville, CA)
Assignee: Corn Products Development, Inc.
C12P19/56C12N15/81C12Y106/02004C12Y114/14C12Y205/01029C12Y402/03019C12Y505/01012
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Quick Facts
Patent No.
US 12,460,239
App. No.
18/767,621
Granted
Nov 4, 2025
Kind
B2
Abstract

Provided herein are genetically modified host cells, compositions, and methods for improved production of steviol glycosides. The host cells are genetically modified to contain a heterologous nucleic acid that expresses novel and optimized variants of UGT76G1. The host cell further contains one or more heterologous nucleotide sequence encoding further enzymes of a pathway capable of producing one or more steviol glycosides in the host cell. The host cells, compositions, and methods described herein provide an efficient route for the heterologous production of rebaudioside M.

Claims (27)

1 . A method for producing one or more steviol glycosides comprising

culturing a population of host cells in a medium with a carbon source under conditions suitable for making one or more steviol glycosides to yield a culture broth; and

recovering the one or more steviol glycosides from the culture broth,

wherein the host cells comprise:

(i) a variant UDP-glycosyltransferase polypeptide comprising the amino acid sequence of SEQ ID NO: 1 with one or more amino acid substitutions, wherein the variant UDP-glycosyltransferase polypeptide produces a higher ratio of RebM/RebD compared to the UDP-glycosyltransferase polypeptide of SEQ ID NO: 1, and wherein the one or more amino acid substitutions are selected from the group consisting of (L85H), (L200G), (L379Q), (H155I), (H155V), (L200K), (V20G), (L200C), (A125F), (H155G), (H155T), (H155L), (L200E), (L200M), and (L379M);

(ii) a variant UDP-glycosyltransferase polypeptide comprising the amino acid sequence of SEQ ID NO: 1 with two or more amino acid substitutions, wherein the variant UDP-glycosyltransferase polypeptide produces a higher ratio of RebM/RebD compared to the UDP-glycosyltransferase polypeptide comprising SEQ ID NO: 1, and wherein the two or more amino acid substitutions are selected from the group consisting of (H155V, L200M), (H155G, L200E), (L85H, A125F, H155G, L200M), (A125F,H155G,L200K,L379M), (L200K,L379Q), (A125F,H155T,L200M), (L85H,L200M,L379Q), (L85H,H155G,L200G,L379M), (L85H,A125F,H155V,L200M), (L85H,A125F,H155I,L200C,L379Q), (A125F,H155L,L200E,L379Q), (H155T,L200G,L379M), (H155L,L200M), (H155V,L200C), (L85H,H155T,L200G), (L85H,H155I,L200C,L379Q), (H155I,L200M,L379Q), (L85H,A125F,H155T,L200E,L379M), (L85H,H155T,L200M), (H155V,L200C,L379M), (L85H,A125F,L200M,L379M), (H155V,L379Q), (L85H,L200K,L379M), (L85H,A125F,H155L,L200M,L379Q), (L85H,A125F,H155I,L200K,L379M), (H155I,L379M), (L200M,L379M), (L85H,H155V,L200G,L379Q), (H155T,L200E), (H155V,L379M), (A125F,H155I,L379M), (H155V,L200C,L379Q), (L85H,H155G,L200K,L379Q), (A125F,H155G,L200M,L379M), (H155T,L200E,L379Q), (L200K,L379G), (L85H,H155L,L200C,L379M), (A125F,H155I,L200K,L379M), (L85H,H155V,L200M), (L85H,H155V,L200E,L379V), (L85H,H155V,L200M,L379Q), (L85H,L200M,L379M), (H155V,L200E,L379M), (L85H,H155I,L200M,L379M), (L85H,H155I,L200M,L379Q), (L85H,A125F,H155T,L200C,L379M), (L85H,A125F,H155L,L200K,L379M), (V20L,L85M,A125P), (L85H,H155L,L200K,L379M), (L85H,H155V,L200C,L379Q), (H155I,L200K,L379M), (L85H,H155L,L200C), (L85H,A125F,H155L,L379M), (L85H,H155I,L200E,L379M), (L85H,A125F,H155G,L200M,L379M), (L85H,H155L,L200E,L379M), (H155L,L200C), (L85H,H155L,L200C,L379E), (L85H,H155V,L200K,L379Q), (H155T,L200K,L379M), (H155V,L200G,L379M), (L85H,A125F,H155V,L200M,L379M), (A125F,H155G,L200C,L379M), (A125F,H155L,L379M), (L85H,H155L,L200G,L379M), (L85H,H155L,L200E,L379Q), (H155G,L200E,L379M), (H155L,L200C,L379Q), (L85H,H155V,L379M), (L85H,A125F,H155I,L200C,L379M), (H155G,L200C,L379M), (A125F,H155V,L200M,L379M), and (H155L,L200E); or

(iii) a nucleic acid encoding the variant UDP-glycosyltransferase polypeptide of (i) or (ii);

wherein the host cells are capable of producing one or more steviol glycosides.

2 . The method of claim 1 , wherein the one or more steviol glycosides are selected from the group consisting of RebA, RebB, RebD, RebE, and RebM.

3 . The method of claim 1 , wherein the host cells comprise one or more nucleic acids encoding one or more enzymes of a pathway for making a steviol glycoside.

4 . The method of claim 1 , wherein the one or more steviol glycosides is RebM.

5 . The method of claim 3 , wherein the one or more steviol glycosides is RebM.

6 . The method of claim 1 , wherein the host cells further comprise a nucleic acid encoding a geranyl diphosphate.

7 . The method of claim 1 , wherein the host cells further comprise a nucleic acid encoding a copalyl diphosphate synthase.

8 . The method of claim 1 , wherein the host cells further comprise a nucleic acid encoding an ent-kaurene synthase.

9 . The method of claim 1 , wherein the host cells further comprise a nucleic acid encoding a kaurenoic acid 13-hydroxylase.

10 . The method of claim 1 , wherein the host cells further comprise a nucleic acid encoding a kaurene oxidase.

11 . The method of claim 1 , wherein the host cells further comprise a nucleic acid encoding a cytochrome P450 reductase.

12 . The method of claim 1 , wherein the host cells further comprise a nucleic acid encoding one or more uridine 5′-disphosphate-dependent glycosyltransferases.

13 . The method of claim 1 , wherein the host cells further comprise one or more nucleic acids encoding a geranyl diphosphate synthase, a copalyl diphosphate synthase, an ent-kaurene synthase, a kaurenoic acid 13-hydroxylase, a kaurene oxidase, a cytochrome P450 reductase, a UGT40087, UGT74G1, UGT85C2, EUGT11, and UGT91D.

14 . The method of claim 1 , wherein the host cells are selected from the group consisting of bacterial cells, yeast cells, algal cells, insect cells, and plant cells.

15 . The method of claim 13 , wherein the host cells are selected from the group consisting of bacterial cells, yeast cells, algal cells, insect cells, and plant cells.

16 . The method of claim 1 , wherein the host cells are yeast cells.

17 . The method of claim 13 , wherein the host cells are yeast cells.

18 . The method of claim 16 , wherein the host cells are Saccharomyces cerevisiae cells.

19 . The method of claim 17 , wherein the host cells are Saccharomyces cerevisiae cells.

20 . The method of claim 19 , wherein the one or more steviol glycosides is RebM.

Assignments (4)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 27, 2025
From: LUND, SEAN; WICHMANN, GALE
To: AMYRIS, INC.
Reel/Frame 071542/0697 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 4, 2025
From: AMYRIS, INC.
To: CORN PRODUCTS DEVELOPMENT, INC.
Reel/Frame 071314/0668 →
RELEASE OF SECURITY INTEREST Recorded May 29, 2025
From: EUAGORE, LLC
To: AMYRIS, INC.
Reel/Frame 071260/0237 →
SECURITY INTEREST Recorded Dec 5, 2024
From: AMYRIS, INC.
To: EUAGORE, LLC
Reel/Frame 069502/0957 →
Continuity (4)
Division 18368369 · Sep 14, 2023
Continuation 16871324 · May 11, 2020
Provisional Application 62846909 · May 13, 2019
Related Publication 20250313877A1 · Oct 9, 2025
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