Transformed Synechococcus elongatus having capability of producing biodiesel from carbon dioxide and method for producing biodiesel using the same

Patent metadata

Field	Value
Publication number	US-10036046-B2
Application number	US-201615299188-A
Country	US
Kind code	B2
Filing date	Oct 20, 2016
Priority date	Oct 22, 2015
Publication date	Jul 31, 2018
Grant date	Jul 31, 2018

Abstract

The present disclosure discloses a transformed Synechococcus elongatus strain capable of producing biodiesel directly from carbon dioxide and a method for producing biodiesel and a method for removing carbon dioxide using the same. In an aspect, the transformed Synechococcus elongatus strain of the present disclosure can produce biodiesel in large scale using carbon dioxide as a carbon source. The Synechococcus elongatus strain is environment-friendly because it can be used to remove or reduce carbon dioxide in the atmosphere. The strain of the present disclosure is advantageous in that it can produce biodiesel in large scale because it grows faster and exhibits excellent carbon dioxide fixation capability as compared to other photosynthetic microorganisms.

First claim

What is claimed is:

1. A Synechococcus elongatus strain comprising: a gene encoding an enzyme which produces acetaldehyde from pyruvate; a gene encoding an enzyme which produces ethanol from acetaldehyde; and a gene encoding an enzyme which produces biodiesel from acyl-coenzyme A and ethanol,
wherein the enzyme which produces acetaldehyde from pyruvate is pyruvate decarboxylase, the enzyme which produces ethanol from acetaldehyde is alcohol dehydrogenase and the enzyme which produces biodiesel from acyl-coenzyme A and ethanol is wax-ester synthase, and
wherein the pyruvate decarboxylase gene comprises the nucleotide sequence of SEQ ID NO: 1, the alcohol dehydrogenase gene comprises the nucleotide sequence of SEQ ID NO: 2, and the wax-ester synthase gene comprises the nucleotide sequence of SEQ ID NO: 3 or SEQ ID NO: 4.

2. The Synechococcus elongatus strain according to claim 1 , wherein the strain further comprises a gene encoding an enzyme which produces a free fatty acid from an acyl-acyl carrier protein (acyl-ACP),
wherein the enzyme which produces a free fatty acid from an acyl-acyl carrier protein (acyl-ACP) is thioesterase,
wherein the thioesterase gene comprises the nucleotide sequence of SEQ ID NO: 5.

3. The Synechococcus elongatus strain according to claim 1 , wherein the strain is transformed with a first vector comprising a sequence derived from a pdc (pyruvate decarboxylase) gene of Zymomonas mobilis and a sequence derived from an adh (alcohol dehydrogenase) gene of Zymomonas mobilis ; and a second vector comprising a sequence derived from an atfA gene of Acinetobacter sp. or a Ws2 gene of Marinobacter hydrocarbonoclasticus.

4. The Synechococcus elongatus strain according to claim 3 , wherein the strain is further transformed with a third vector comprising a sequence derived from a tesA (thioesterase) gene of E. coli.

5. The Synechococcus elongatus strain according to claim 3 , wherein the first vector is inserted at the neutral site II (NSII) of the parent strain Synechococcus elongatus and the second vector is inserted at the neutral site I (NSI) of the parent strain Synechococcus elongatus.

6. The Synechococcus elongatus strain according to claim 4 , wherein the third vector is inserted at the FadE site of the parent strain Synechococcus elongatus.

7. The Synechococcus elongatus strain according to claim 3 ,
wherein the first vector comprises, in sequence, a kanamycin resistance gene, a lacI (lactose) repressor, a trc promoter, the gene encoding pyruvate decarboxylase and the gene encoding alcohol dehydrogenase; or
wherein the second vector comprises, in sequence, a spectinomycin resistance gene, a lacI repressor, a trc promoter and the gene encoding wax-ester synthase.

8. The Synechococcus elongatus strain according to claim 4 , wherein the third vector comprises, in sequence, a chloramphenicol resistance gene, a lacI repressor, a trc promoter and the gene encoding thioesterase.

9. The Synechococcus elongatus strain according to claim 3 , wherein the first vector comprises the nucleotide sequence of SEQ ID NO 6 and the second vector comprises the nucleotide sequence of SEQ ID NO 7 or SEQ ID NO 8.

10. The Synechococcus elongatus strain according to claim 4 , wherein the third vector comprises the nucleotide sequence of SEQ ID NO 9.

11. The Synechococcus elongatus strain according to claim 4 , wherein the strain transformed with the vectors is derived from the parent strain Synechococcus elongatus PCC7942 (ATCC 33912).

12. The Synechococcus elongatus strain according to claim 1 , wherein the biodiesel is a fatty acid ethyl ester (FAEE).

13. The Synechococcus elongatus strain according to claim 1 , wherein the strain is a Synechococcus elongatus strain with an accession number KCTC 12883BP or an accession number KCTC 12884BP.

14. The Synechococcus elongatus strain according to claim 1 , wherein the strain absorbs and fixes carbon dioxide.

15. A method for producing biodiesel comprising culturing the Synechococcus elongatus strain according to claim 14 .

16. A method for removing or reducing carbon dioxide comprising culturing the Synechococcus elongatus strain according to claim 14 .

Assignees

• Korea Inst Sci & Tech

Inventors

• Woo Han Min
• Lee Hyun Jeong
• Um Youngsoon
• Gong Gyeong Taek
• Lee Sun Mi
• Kim Yunje

Classifications

• C12Y101/01001

  Alcohol dehydrogenase (1.1.1.1) · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12N9/16

  acting on ester bonds (3.1) · CPC title

• C12Y401/01001

  Pyruvate decarboxylase (4.1.1.1) · CPC title

• C12N9/88

  Lyases (4.) · CPC title