Glycolipid production

Abstract

The present invention relates to a microbial cell that produces at least one lipid represented by the general formula II: 【Chemical 1】 JPEG2026519410000018.jpg53170(wherein R 1 and R 2 each independently have the same or different organic radicals having 5 to 13 carbon atoms.) and is a non-pathogenic cell genetically modified to increase the heterologous expression of the following enzymes: - Enzyme E1: a 3-(3-hydroxyalkanoyloxy)alkanoic acid (HAA) synthase comprising SEQ ID NO: 1, SEQ ID NO: 7, SEQ ID NO: 11, or SEQ ID NO: 15, or a variant thereof having 60% sequence identity with SEQ ID NO: 1, SEQ ID NO: 7, SEQ ID NO: 11, or SEQ ID NO: 15, and - Enzyme E2: a glycosyltransferase (EC 2.4) comprising SEQ ID NO: 2, SEQ ID NO: 8, or SEQ ID NO: 12, or a variant thereof having 60% sequence identity with SEQ ID NO: 2, SEQ ID NO: 8, or SEQ ID NO: 12 The present invention relates to a cell which is a non-pathogenic cell genetically modified to increase the heterologous expression of the above enzymes.​

Claims

1. From at least one carbon substrate, General Formula II: 【Chemistry 1】 General formula II (In the formula, R １ and R ２ Each of these independently possesses the same or different organic radicals having 5 to 13 carbon atoms. A microbial cell that produces at least one lipid represented by, Compared to wild-type cells, the following enzymes: - Enzyme E １ : 3-(3-hydroxyalkanoyloxy)alkanoic acid (HAA) synthase containing SEQ ID NO: 1, SEQ ID NO: 7, SEQ ID NO: 11, or SEQ ID NO: 15, or a variant thereof having 60% sequence identity with SEQ ID NO: 1, SEQ ID NO: 7, SEQ ID NO: 11, or SEQ ID NO: 15, and - Enzyme E ２ : Glycosyltransferase (EC2.4) containing SEQ ID NO: 2, SEQ ID NO: 8, or SEQ ID NO: 12, or a variant thereof having 60% sequence identity with SEQ ID NO: 2, SEQ ID NO: 8, or SEQ ID NO: 12 Cells that are non-pathogenic cells that have been genetically modified to increase heterologous expression of a particular gene.

2. The cell according to claim 1, wherein R in the lipid represented by the general formula II is a saturated alkyl radical.

3. The cell according to claim 2, wherein the alkyl radical is selected from the group consisting of pentyl, heptyl, nonyl, undecyl, and tridecyl.

4. From the aforementioned carbon substrate, General Formula I: 【Chemistry 2】 General formula I (In the formula, R １ and R ２ Each of these independently possesses the same or different organic radicals having 5 to 13 carbon atoms. A cell according to any one of claims 1 to 3, which further produces a lipid represented by [the specified formula].

5. The cell according to claim 4, wherein R in the lipid represented by the general formula I is a saturated alkyl radical.

6. The cell according to any one of claims 1 to 5, wherein the carbon source is selected from the group consisting of glucose, dextrose, sucrose, xylose, mannose, galactose, arabinose, monosaccharides, polysaccharides, cellulose, hemicellulose, vegetable oil, animal fat, fatty acids, fatty acid esters, carbonaceous gases, alkanes, glycerol, acetate, ethanol, and methanol.

7. A cell according to any one of claims 1 to 6, selected from the group consisting of Acinetobacter, Bacillus, Brevibacterium, Burkholderia, Chlorella, Clostridium, Corynebacterium, Cyanobakterien, Escherichia, Pseudomonas, Klebsiella, Salmonella, Rhizobium, Saccharomyces, Pichia, and Nostoc.

8. A cell according to any one of claims 1 to 7, selected from the group consisting of Bacillus subtilis, Burkholderia thailandensis, Corynebacterium glutamicum, E. coli, Klebsiella oxytoca, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas stutzeri, Rhizobium meliloti, Saccharomyces cerevisiae, and Pichia pastoris.

9. General formula II and / or general formula I: 【Transformation 3】 General formula II General formula I (In the formula, R １ and R ２ Each of these independently possesses the same or different organic radicals having 5 to 13 carbon atoms. A method for producing at least one lipid represented by, A method comprising the step of bringing at least one cell according to any one of claims 1 to 8 into contact with at least one carbon source.

10. The method according to claim 9, wherein the carbon source is selected from the group consisting of glucose, dextrose, sucrose, xylose, mannose, galactose, arabinose, monosaccharides, polysaccharides, cellulose, hemicellulose, vegetable oil, animal fat, fatty acids, fatty acid esters, carbonaceous gases, alkanes, glycerol, acetate, ethanol, and methanol.

11. General formula I and / or general formula II: 【Chemistry 4】 General formula II General formula I (wherein, R １ and R ２ each independently has the same or different organic radicals having 5 to 13 carbon atoms.) Use of cells according to any one of claims 1 to 8 for producing at least one lipid represented by

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