Modified photosynthetic microorganisms with reduced glycogen and their use in producing carbon-based products
This disclosure describes genetically modified photosynthetic microorganisms, including Cyanobacteria, that contain one or more mutations or deletions in a glycogen biosynthesis or storage pathway, which accumulate a reduced amount of glycogen as compared to a wild type Cyanobacterium, and which are capable of producing an increased amount of lipids and/or fatty acids. In certain embodiments, the genetically modified photosynthetic microorganisms also contain one or more exogenous genes encoding a diacyglycerol acyltransferase, a phosphatidate phosphatase, and/or an acetyl-CoA carboxylase, and which are capable of producing increased amounts of lipids or fatty acids and/or synthesizing triglycerides.
1. A modified photosynthetic microorganism that accumulates a reduced amount of glycogen as compared to the wild type photosynthetic microorganism,
wherein said modified photosynthetic microorganism has reduced expression, reduced activity, or mutation of a glucose-1-phosphate adenyltransferase (glgC) gene as compared to the wild type photosynthetic microorganism, wherein said glgC gene comprises one or more mutations or complete or partial gene deletions,
wherein said modified photosynthetic microorganism comprises one or more introduced polynucleotides encoding diacylglycerol acyltransferase (DGAT),
wherein said modified photosynthetic microorganism accumulates an increased amount of triglycerides, wax esters, or both as compared to a photosynthetic microorganism having the introduced polynucleotide(s) encoding DGAT without reduced expression or mutation of the glgC gene, and
wherein said photosynthetic microorganism is a Cyanobacterium.
2. The modified photosynthetic microorganism of claim 1 , wherein said modified photosynthetic microorganism accumulates a reduced amount of glycogen under stress conditions as compared to the wild type photosynthetic microorganism.
3. The modified photosynthetic microorganism of claim 1 , wherein said Cyanobacterium is a Synechococcus elongatus.
4. The modified Cyanobacterium of claim 3 , wherein the Synechococcus elongatus is strain PCC 7942.
5. The modified Cyanobacterium of claim 4 , wherein the Cyanobacterium is a salt tolerant variant of Synechococcus elongatus strain PCC 7942.
6. The modified photosynthetic microorganism of claim 1 , wherein said Cyanobacterium is Synechococcus sp. PCC 7002 or Synechocystis sp. PCC 6803.
7. The modified photosynthetic microorganism of claim 1 , wherein said one or more polynucleotides encoding DGAT are present in one or more expression vectors.
8. The modified photosynthetic microorganism of claim 1 , wherein said glgC gene is an endogenous glgC gene that hybridizes to at least one of SEQ ID NOs: 59, 61, 63, 65, 67, 69, 71 or 73 under low stringency conditions.
9. A method of producing a modified photosynthetic microorganism that accumulates a reduced amount of glycogen as compared to the wild type photosynthetic microorganism, comprising,
modifying a photosynthetic microorganism to have a reduced level of expression, reduced activity, or mutation of a glucose-1-phosphate adenyltransferase (glgC) gene as compared to the level of expression of the glgC gene in a wild type photosynthetic microorganism by introducing one or more mutations or complete or partial gene deletions into said glgC gene, and
modifying the photosynthetic microorganism to include one or more introduced polynucleotides encoding diacylglycerol acyltransferase (DGAT),
wherein said modified photosynthetic microorganism accumulates an increased amount of triglycerides, wax esters, or both as compared to a photosynthetic microorganism having the introduced polynucleotide(s) encoding DGAT without reduced expression or mutation of the glgC gene, and
wherein said photosynthetic microorganism is a Cyanobacterium.
10. The method of claim 9 , wherein said glgC gene is an endogenous glgC gene that hybridizes to at least one of SEQ ID NOs: 59, 61, 63, 65, 67, 69, 71 or 73 under low stringency conditions.
11. A method of producing a triglyceride, a wax ester, or both, comprising cultivating a modified photosynthetic microorganism in a suitable media to produce said triglyceride, wax ester, or both, said modified photosynthetic microorganism having:
reduced expression, reduced activity, or mutation of a glucose-l-phosphate adenyltransferase (glgC) gene wherein said glgC gene comprises one or more mutations or complete or partial gene deletions,
wherein said modified photosynthetic microorganism comprises one or more introduced polynucleotides encoding diacylglycerol acyltransferase (DGAT),
wherein said modified photosynthetic microorganism accumulates an increased amount of triglycerides, wax esters, or both as compared to a photosynthetic microorganism having the introduced polynucleotide(s) encoding DGAT without reduced expression or mutation of said glgC gene, and
wherein said photosynthetic microorganism is a Cyanobacterium.
12. The method of claim 11 , wherein said glgC gene is an endogenous glgC gene that hybridizes to at least one of SEQ ID NOs: 59, 61, 63, 65, 67, 69, 71 or 73 under low stringency conditions.