Biosynthetic production of gamma-lactones

Patent metadata

Field	Value
Publication number	US-11549131-B2
Application number	US-202117150774-A
Country	US
Kind code	B2
Filing date	Jan 15, 2021
Priority date	Jul 17, 2018
Publication date	Jan 10, 2023
Grant date	Jan 10, 2023

Abstract

Provided herein are methods for making gamma lactones comprising reacting a carboxylic acid substrate with a heterologous cytochrome P450 (CYP450) protein with carboxylic acid 4-hydroxylase activity.

First claim

The invention claimed is:

1. A method for producing a C4-C20 gamma lactone, the method comprising:
providing a reaction mixture comprising a recombinant CYP450 protein having carboxylic acid 4-hydroxylase activity and a C4-C20 carboxylic acid substrate to provide a 4-hydroxy C4-C20 carboxylic acid; and
subjecting the 4-hydroxy C4-C20 carboxylic acid to acidic conditions to produce the C4-C20 gamma lactone,
wherein the recombinant CYP450 protein comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 1.

2. The method of claim 1 , wherein the recombinant CYP450 protein is expressed by a transformed isolated host cell in the reaction mixture.

3. The method of claim 2 , wherein the recombinant CYP450 protein comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 1.

4. The method of claim 1 , wherein the recombinant CYP450 protein comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 1.

5. The method of claim 1 , wherein the C4-C20 carboxylic acid substrate comprises a C4-C20 carboxylic acid; a salt of a C4-C20 carboxylic acid; an ester of a C4-C20 carboxylic acid; a mono-, di- or triglyceride of a C4-C20 carboxylic acid; or a combination thereof.

6. The method of claim 1 , wherein the C4-C20 carboxylic acid comprises a straight-chain C4-C20 carboxylic acid, a branched C4-C20 carboxylic acid, a saturated C4-C20 carboxylic acid, an unsaturated C4-C20 carboxylic acid, or a combination thereof.

7. The method of claim 4 for producing a C4-C20 gamma lactone selected from γ-heptalactone, γ-octalactone, γ-nonalactone, γ-decalactone, γ-undecalactone, γ-dodecalactone, γ-tridecalactone, and γ-tetradecalactone, wherein the C4-C20 carboxylic acid substrate is selected from heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, and tetradecanoic acid.

8. The method of claim 1 for producing a C4-C20 gamma lactone selected from γ-undecalactone and γ-undecenolactone, wherein the C4-C20 carboxylic acid substrate is selected from undecanoic acid and undecenoic acid.

9. The method of claim 1 for producing a C4-C20 gamma lactone selected from γ-hexalactone, γ-heptalactone, γ-decalactone, γ-undecalactone, γ-dodecalactone, γ-tridecalactone, and γ-tetradecalactone, wherein the C4-C20 carboxylic acid substrate is selected from hexanoic acid, heptanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, and tetradecanoic acid.

10. The method of claim 2 , wherein the transformed isolated host cell is selected from a bacterial cell and a yeast cell.

11. The method of claim 2 , wherein the transformed isolated host cell is selected from a group comprising of Escherichia; Salmonella; Bacillus; Acinetobacter; Streptomyces; Corynebacterium; Methylosinus; Methylomonas; Rhodococcus; Pseudomonas; Rhodobacter; Synechocystis; Saccharomyces; Zygosaccharomyces; Kluyveromyces; Candida; Hansenula; Debaryomyces; Mucor; Pichia; Torulopsis; Aspergillus; Arthrobotlys; Brevibacteria; Microbacterium; Arthrobacter; Citrobacter; Klebsiella; Mucor; Pantoea; Corynebacterium ; and Clostridium.

12. The method of claim 1 , wherein the recombinant CYP450 protein comprises the amino acid sequence of SEQ ID NO: 1.

Assignees

• Conagen Inc

Inventors

• Chen Hui
• Yu Oliver

Classifications

• C12P17/04 Primary

  containing a five-membered hetero ring, e.g. griseofulvin {, vitamin C} · CPC title

• C12N15/52 Primary

  Genes encoding for enzymes or proenzymes · CPC title