Methods of using capsaicin synthase for the microbial production of capsaicinoids

Patent metadata

Field	Value
Publication number	US-9951358-B2
Application number	US-201515111901-A
Country	US
Kind code	B2
Filing date	Jan 16, 2015
Priority date	Jan 17, 2014
Publication date	Apr 24, 2018
Grant date	Apr 24, 2018

Abstract

A biosynthetic method of making a capsaicinoid including expressing a first gene product of CS/AT3/Pun1 in a cellular system, growing the cellular system in a medium, and collecting the capsaicinoid. The biosynthetic method further includes expressing a second gene product of ACS1 and expressing a third gene product of pAMT in the cellular system.

First claim

The claims are the following:

1. A bioconversion method of making a capsaicinoid comprising:
(a) expressing a first gene product of CS/AT3/Pun1 in a mixture;
(b) providing a first substrate to the mixture;
(c) expressing a second gene product of ACS1 in said mixture;
(d) providing a second substrate to the mixture;
(e) collecting the capsaicinoid; and,
wherein the CS/AT3/Pun1 comprises the DNA sequence of SEQ ID No. 1 wherein the gene products are first expressed within a cellular system.

2. The bioconversion method of making a capsaicinoid of claim 1 , wherein the CS/AT3/Pun1 DNA sequence is derived from ghost chili pepper.

3. The bioconversion method of making a capsaicinoid of claim 1 , wherein the first substrate is an activated fatty acid selected from the group consisting of 8-methyl-6-nonenoyl-CoA, 8-methyl nonanoyl-CoA, octanoyl-CoA, decanoyl-CoA, other medium to long-chain acyl CoAs and a combination thereof.

4. The bioconversion method of making a capsaicinoid of claim 1 , wherein the ACS1 is derived from ghost chili pepper.

5. The bioconversion method of making a capsaicinoid of claim 1 , wherein the second substrate is a fatty acid selected from the group consisting of 8-methyl-6-nonenoic acid, 8-methyl nonanoic acid, octanoic acid, decanoic acid, other medium-to long-chain fatty acids and a combination thereof.

6. The bioconversion method of making a capsaicinoid acid of claim 1 , wherein the cellular system comprises a microorganism selected from the group consisting of bacteria, yeast and a combination thereof.

7. The bioconversion method of making a capsaicinoid of claim 1 , further comprising providing a third substrate vanillylamine.

8. The bioconversion method of making a capsaicinoid acid of claim 7 , wherein the cellular system comprises a microorganism selected from the group consisting of bacteria, yeast and a combination thereof.

9. A bioconversion method of making a capsaicinoid comprising:
(a) expressing a gene product of CS/AT3/Pun1 in a cellular system;
(b) providing fatty acid-CoA;
(c) expressing a second gene product of ACS1 in said cellular system;
(d) providing vanillylamine;
(e) growing the cellular system in a medium;
(f) collecting the capsaicinoid; and,
wherein the of CS/AT3/Pun1 comprises the DNA sequence of SEQ ID No. 1 and wherein the gene products are expressed within said cellular system.

10. The bioconversion method of making a capsaicinoid acid of claim 9 , wherein the cellular system comprises a microorganism selected from the group consisting of bacteria, yeast and a combination thereof.

Assignees

• Conagen Inc

Inventors

• Chen Hui
• Wang Hongxue
• Yu Oliver

Classifications

• C12N9/1096

  transferring nitrogenous groups (2.6) · CPC title

• C12N9/20

  Triglyceride splitting, e.g. by means of lipase · CPC title

• C12Y602/01001

  Acetate-CoA ligase (6.2.1.1) · CPC title

• A61K31/165 Primary

  having aromatic rings, e.g. colchicine, atenolol, progabide · CPC title

• C12N9/1029

  transferring groups other than amino-acyl groups (2.3.1) · CPC title