# US 10,246,695 B2 · Methyl butenol synthase

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- Number: US 10,246,695 B2
- Document ID: US10246695B2
- Status: Granted
- Kind: B2
- Application number: 15/797,632
- Filed: 2017-10-30
- Published: 2018-05-31
- Granted: 2019-04-02
- Earliest claimed: 2009-05-22
- Assignees: Board of Trustees of Michigan State University
- Applicants: Board of Trustees of Michigan State University
- Inventors: Dennis Gray; Thomas D Sharkey
- Web page: https://patentgrep.com/patent/US10246695B2

## CPC

- C12N 9/88: Lyases (4.)
- C12P 7/04: acyclic
- C12P 7/16: Butanols
- Y02E 50/10: Biofuels, e.g. bio-diesel
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)}
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

The present invention provides novels genes encoding methyl butenol (MBO) synthase, methyl butenol synthases and their use in methyl butenol production.

## Claims

### Claim 1

An isolated and purified methyl butenol (MBO) synthase cDNA molecule, wherein the MBO synthase cDNA molecule comprising any one of SEQ ID NOs: 12-17 or at least 95% identity thereto, or a cDNA molecule encoding MBO synthase polypeptide comprising any one of SEQ ID NOs: 29-34.

### Claim 2 (depends on 1)

An expression vector comprising a MBO synthase cDNA molecule of claim 1.

### Claim 3 (depends on 1)

An isolated prokaryotic or eukaryotic host cell transformed with a MBO synthase cDNA molecule of claim 1.

### Claim 4 (depends on 2)

A prokaryotic or eukaryotic host cell transformed with the vector of claim 2.

### Claim 5 (depends on 3)

The host cell of claim 3, wherein the transformed cell expresses MBO synthase.

### Claim 6

An isolated and purified methyl butenol (MBO) synthase cDNA, wherein the MBO synthase cDNA comprising any one of SEQ ID NOs: 12-17 or a MBO synthase cDNA molecule encoding MBO synthase polypeptide comprising any one of SEQ ID NOs: 29-34.

### Claim 7 (depends on 1)

A method to produce methyl butenol comprising transforming a host cell with a cDNA molecule of claim 1 and expressing said molecule in a host cell so as to yield methyl butenol.

### Claim 8 (depends on 7)

The method of claim 7, wherein said host cell is a strain of yeast, bacteria, cyanobacteria, eukaryotic micro algae or a combination thereof.

### Claim 9 (depends on 8)

The method of claim 8, wherein said yeast is Saccharomyces cerevisiae.

### Claim 10 (depends on 8)

The method of claim 8, wherein said bacteria is a C5- or C6-fermentative organism.

### Claim 11 (depends on 10)

The method of claim 10, wherein said C5-fermenative organism is a species of Zymomonas.

### Claim 12 (depends on 8)

The method of claim 8, wherein said bacteria is Klebsiella oxytoca.

### Claim 13 (depends on 8)

The method of claim 8, wherein said cyanobacteria is Synechococcussp., Synechocystis sp., or Anabaena sp.

### Claim 14 (depends on 8)

The method of claim 8, wherein said eukaryotic micro algae is Chlorella sp., Scenedesnius sp., Bracteococcus sp. or Chlamydomonus sp.