# US 9,518,277 B2 · Genetically engineered microbial strains including Chlorella protothecoides lipid pathway genes

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- Number: US 9,518,277 B2
- Document ID: US9518277B2
- Status: Granted
- Kind: B2
- Application number: 14/099,704
- Filed: 2013-12-06
- Published: 2014-06-26
- Granted: 2016-12-13
- Earliest claimed: 2012-12-07
- Assignees: TerraVia Holdings, Inc.
- Applicants: TerraVia Holdings, Inc.
- Inventors: Scott Franklin; Shane Brubaker; George N. Rudenko; Riyaz Bhat; Matthew Shoa-Azar; Aravind Somanchi; Aren Ewing
- Web page: https://patentgrep.com/patent/US9518277B2

## CPC

- C12P 7/6463: obtained from glyceride producing microorganisms, e.g. single cell oil
- C12N 15/52: Genes encoding for enzymes or proenzymes
- 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

Genetically engineered microbial, e.g., Chlorella protothecoides, cells producing microbial oils are useful as a food additive and a source of renewable fuels and industrial chemicals. Lipid biosynthesis genes for Auxenochlorella protethecoides are disclosed that are useful for increasing lipid production and altering fatty acid and triacylglycerol profiles in recombinant microrganisms including cells of the genus Auxenochlorella or Prototheca.

## Claims

### Claim 1

A recombinant cell transformed with an expression vector that comprises an exogenous nucleic acid encoding the polypeptide of SEQ ID NO:49127.

### Claim 2 (depends on 1)

The recombinant cell of claim 1, wherein the nucleic acid is operably linked to a promoter.

### Claim 3 (depends on 1)

The recombinant cell of claim 1, wherein the nucleic acid is operably linked to an untranslated control element.

### Claim 4 (depends on 1)

The recombinant cell of claim 1, wherein the nucleic acid is operably linked to a sequence encoding a targeting peptide.

### Claim 5 (depends on 4)

The recombinant cell of claim 4, wherein the targeting peptide is a transit peptide selected from the group of a plastidial targeting peptide and a mitochondrial targeting peptide.

### Claim 6 (depends on 1)

The recombinant cell of claim 1, wherein the nucleic acid is a DNA molecule.

### Claim 7 (depends on 1)

The recombinant cell of claim 1, wherein the vector further comprises a nucleic acid that encodes a sucrose invertase.

### Claim 8 (depends on 1)

The recombinant cell of claim 1, wherein the cell is a cell of the genus Prototheca or Chlorella.

### Claim 9 (depends on 8)

The recombinant cell of claim 8, wherein the cell is a cell of the species Chlorella protothecoides.

### Claim 10 (depends on 9)

A method for obtaining microbial oil comprising culturing the recombinant Chlorella protothecoides cell of claim 9 under conditions such that oil is produced.