# US 11,306,335 B2 · Genes and proteins for aromatic polyketide synthesis

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- Number: US 11,306,335 B2
- Document ID: US11306335B2
- Status: Granted
- Kind: B2
- Application number: 16/917,362
- Filed: 2020-06-30
- Published: 2020-10-15
- Granted: 2022-04-19
- Earliest claimed: 2010-04-15
- Assignees: National Research Council of Canada; University of Saskatchewan
- Applicants: National Research Council of Canada; University of Saskatchewan
- Inventors: Jonathan E. Page; Steve Gagne
- Web page: https://patentgrep.com/patent/US11306335B2

## CPC

- C12P 17/06: containing a six-membered hetero ring, e.g. fluorescein
- C12N 9/1025: {Acyltransferases (2.3)}
- C12N 15/52: Genes encoding for enzymes or proenzymes
- C12N 15/70: Vectors or expression systems specially adapted for E. coli
- C12N 15/8243: {involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine}
- C12P 7/22: aromatic
- C12P 7/42: Hydroxy-carboxylic acids
- C12Y 203/01206: 3,5,7-Trioxododecanoyl-CoA synthase (2.3.1.206)
- C12Y 404/01026: Olivetolic acid cyclase (4.4.1.26)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## Abstract

Nucleic acid molecules encoding polypeptides having polyketide synthase activity have been identified and characterized. Expression or over-expression of the nucleic acids alters levels of cannabinoid compounds in organisms. The polypeptides may be used in vivo or in vitro to produce cannabinoid compounds.

## Claims

### Claim 1

A process of synthesizing a polyketide in an in vitro cell-free reaction, said process comprising:
reacting an acyl carboxylic acid with coenzyme A through the action of an acyl CoA synthetase to form an alkanoyl CoA in the presence of a type III polyketide synthase enzyme and a polypeptide with at least 95% sequence identity to SEQ ID NO: 2 or that is a conservatively substituted amino acid sequence of the sequence set forth in SEQ ID NO: 2 and having polyketide cyclase activity.

### Claim 2 (depends on 1)

The process of claim 1, further comprises reacting the alkanoyl CoA with a malonyl CoA.

### Claim 3 (depends on 2)

The process of claim 2, wherein the alkanoyl CoA is CoA-S—C(O)—(CH2)n—CH3, where n is an integer from 0 to 6.

### Claim 4 (depends on 3)

The process of claim 3, wherein the alkanoyl CoA comprises hexanoyl CoA, butyryl CoA, or octanoyl CoA.

### Claim 5 (depends on 2)

The process of claim 2, wherein the type III polyketide synthase enzyme is polyketide synthase/olivetol synthase.

### Claim 6 (depends on 4)

The process of claim 4, wherein the alkanoyl CoA is hexanoyl CoA.

### Claim 7 (depends on 6)

The process of claim 6, wherein the polyketide is olivetolic acid.

### Claim 8 (depends on 4)

The process of claim 4, wherein the alkanoyl CoA is butyryl CoA.

### Claim 9 (depends on 4)

The process of claim 4, wherein the alkanoyl CoA is octanoyl CoA.