# US 11,028,417 B1 · Isolated codon sequence

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- Number: US 11,028,417 B1
- Document ID: US11028417B1
- Status: Granted
- Kind: B1
- Application number: 16/553,103
- Filed: 2019-08-27
- Granted: 2021-06-08
- Earliest claimed: 2015-04-09
- Assignees: CB Therapeutics, Inc.
- Applicants: CB Therapeutics, INC.
- Inventors: Sher Ali Butt; Jacob Michael Vogan
- Web page: https://patentgrep.com/patent/US11028417B1

## CPC

- C12P 13/02: Amides, e.g. chloramphenicol {or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes (peptides C12P21/00 or C07K)}
- C12N 15/52: Genes encoding for enzymes or proenzymes
- C12P 7/22: aromatic
- C12P 7/42: Hydroxy-carboxylic acids
- C12P 17/06: containing a six-membered hetero ring, e.g. fluorescein
- 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)

## Abstract

The present invention is a method for the biosynthesis of hundreds of compounds, mainly found in the cannabis plant. The starting material for these compounds can be any biological compound that is used/produced in a biological organism from the sugar family starting materials or other low cost raw materials processed via enzymes or within organisms to give final products. These final products include, but are not limited to: cannabinoids, terpenoids, stilbenoids, flavonoids, phenolic amides, lignanamides, spermidine alkaloids, and phenylpropanoids. Specifically, the present invention relates to the regular, modified, or synthetic gene(s) of select enzymes that are processed and inserted into an expression system (for example, a vector, cosmid, BAC, YAC, phage) to produce modified hosts. The modified host is then optimized for efficient production and yield via manipulation, silencing, and amplifying inserted or other genes in the host, leading to an efficient system for product.

## Claims

### Claim 1

An isolated nucleic acid molecule comprising a nucleotide sequence that is at least 95% identical to the nucleotides 5052 to 6671 of SEQ ID NO: 35, wherein the isolated nucleic acid molecule: (i) is non-naturally occurring; (ii) has a nucleotide sequence that is codon-optimized for expression in a host organism and encodes olivetolic acid cyclase (OAC) polypeptide; and (iii) produces olivetolic acid or divarinolic acid in the host organism when inserted and expressed in the host organism, and wherein the host organism is Escherichia coli, Saccharomyces cerevisiae or Pichia pastoris.

### Claim 2

A host organism that comprises an isolated nucleic acid molecule comprising a nucleotide sequence that is at least 95% identical to the nucleotides 5052 to 6671 of SEQ ID NO: 35, wherein the isolated nucleic acid molecule: (i) is non-naturally occurring; (ii) has a nucleotide sequence that is codon-optimized for expression in the host organism and encodes olivetolic acid cyclase (OAC) polypeptide; and (iii) produces olivetolic acid or divarinolic acid in the host organism when inserted and expressed in the host organism, and wherein the host organism is a bacterium, plant alga or fungus.

### Claim 3 (depends on 2)

The host organism of claim 2, wherein the host organism is Escherichia coli, Saccharomyces cerevisiae or Pichia pastoris.

### Claim 4 (depends on 1)

The isolated nucleic acid molecule of claim 1, wherein the nucleic acid molecule comprises a nucleotide sequence that is at least 95% identical to the sequence set forth in SEQ ID NO: 35.

### Claim 5 (depends on 1)

The isolated nucleic acid molecule of claim 1, comprising the nucleotide sequence set forth in the nucleotides 5052 to 6671 of SEQ ID NO: 35.

### Claim 6 (depends on 4)

The isolated nucleic acid molecule of claim 4, comprising the nucleotide sequence set forth in SEQ ID NO: 35.