Microorganism producing polyunsaturated fatty acid and method for producing polyunsaturated fatty acid

A microorganism capable of producing a ω-6 PUFA,

• in which genes encoding respective domains described in the following (a) to (j) having an activity of synthesizing a PUFA (hereinafter referred to as PUFA-PKS activity) have been introduced into a microorganism that does not have a PUFA metabolic pathway:
• (a) a β-ketoacyl-ACP synthase (hereinafter referred to as KS) domain;
• (b) a malonyl-CoA:ACP acyltransferase (hereinafter referred to as MAT) domain;
• (c) an acyl carrier protein (ACP) domain;
• (d) a ketoreductase (hereinafter referred to as KR) domain;
• (e) a PS-DH domain;
• (f) a chain elongation factor (hereinafter, referred to as CLF) domain;
• (g) an acyltransferase (hereinafter referred to as AT) domain;
• (h) a FabA-DH domain;
• (i) an enoyl-ACP reductase (hereinafter referred to as ER) domain;
• and
• (j) a phosphopantetheine transferase (hereinafter referred to as PPT) domain, and
• wherein
• the reactivity of a PS-DH domain with 3-hydroxyhexanoyl ACP in the microorganism is higher than the reactivity of a FabA-DH domain with 3-hydroxyhexanoyl ACP in the microorganism
• wherein the introduced genes are one of the following combinations:
• (i) phoA, phoB, phoC, phoD, epaE, araB,
• (ii) epaA, phoB, phoC, phoD, epaE, araB,
• (iii) epaA, araB, araC, araD, epaE,
• (iv) phoA, araB, araC, araD, epaE, and
• (v) orfA, orfB, orfC, epaE, araB, and
• wherein
• the amino acid sequence of the protein encoded by the arab gene is SEQ ID NO:
• 58,
• the amino acid sequence of the protein encoded by the araC gene is SEQ ID NO:
• 60,
• the amino acid sequence of the protein encoded by the araD gene is SEQ ID NO:
• 62,
• the amino acid sequence of the protein encoded by the phoA gene is SEQ ID NO:
• 66,
• the amino acid sequence of the protein encoded by the phoB gene is SEQ ID NO:
• 68,
• the amino acid sequence of the protein encoded by the phoC gene is SEQ ID NO:
• 70,
• the amino acid sequence of the protein encoded by the phoD gene is SEQ ID NO:
• 72,
• the amino acid sequence of the protein encoded by the epaA gene is SEQ ID NO:
• 74,
• the amino acid sequence of the protein encoded by the epaE gene is SEQ ID NO:
• 82,
• the amino acid sequence of the protein encoded by the orfA gene is SEQ ID NO:
• 84,
• the amino acid sequence of the protein encoded by the orfB gene is SEQ ID NO:
• 86 or 90, and
• the amino acid sequence of the protein encoded by the orfC gene is SEQ ID NO:
• 88 or 92.

A method for producing a ω-6 PUFA or a ω-6 PUFA-containing composition using the following microorganism (I) or (II) capable of producing a ω-6 PUFA:

• (I) a microorganism capable of producing a ω-6 PUFA, in which the reactivity of a PS-DH domain with 3-hydroxyhexanoyl ACP in the microorganism is higher than the reactivity of a FabA-DH domain with 3-hydroxyhexanoyl ACP in the microorganism,
• wherein the microorganism has an anaerobic pathway involving a protein complex having PUFA-PKS activity,
• wherein proteins constituting the protein complex having PUFA-PKS activity are one of the following combinations (i) to (v):
• (i) PhoA, PhoB, PhoC, PhoD, EpaE, AraB,
• (ii) EpaA, PhoB, PhoC, PhoD, EpaE, AraB,
• (iii) EpaA, AraB, AraC, AraD, EpaE,
• (iv) PhoA, AraB, AraC, AraD, EpaE, and
• (v) OrfA, OrfB, OrfC, EpaE, AraB, and
• wherein
• the amino acid sequence of AraB is SEQ ID NO:
• 58,
• the amino acid sequence of AraC is SEQ ID NO:
• 60,
• the amino acid sequence of AraD is SEQ ID NO:
• 62,
• the amino acid sequence of PhoA is SEQ ID NO:
• 66,
• the amino acid sequence of PhoB is SEQ ID NO:
• 68,
• the amino acid sequence of PhoC is SEQ ID NO:
• 70,
• the amino acid sequence of PhoD is SEQ ID NO:
• 72,
• the amino acid sequence of EpaA is SEQ ID NO:
• 74,
• the amino acid sequence of EpaE is SEQ ID NO:
• 82,
• the amino acid sequence of OrfA is SEQ ID NO:
• 84,
• the amino acid sequence of OrfB is SEQ ID NO:
• 86 or 90, and
• the amino acid sequence of OrfC is SEQ ID NO:
• 88 or 92,
• or
• (II) a microorganism capable of producing a ω-6 PUFA, in which genes encoding respective domains described in the following (a) to (j) having a PUFA-PKS activity have been introduced into a microorganism that does not have a PUFA metabolic pathway:
• (a) a KS domain;
• (b) a MAT domain;
• (c) an ACP domain;
• (d) a KR domain;
• (e) a PS-DH domain;
• (f) a CLF domain;
• (g) an AT domain;
• (h) a FabA-DH domain;
• (i) an ER domain;
• and
• (j) a PPT domain,
• wherein the reactivity of a PS-DH domain with 3-hydroxyhexanoyl ACP in the microorganism is higher than the reactivity of a FabA-DH domain with 3-hydroxyhexanoyl ACP in the microorganism
• wherein the introduced genes are one of the following combinations:
• (i) phoA, phoB, phoC, phoD, epaE, araB,
• (ii) epaA, phoB, phoC, phoD, epaE, araB,
• (iii) epaA, araB, araC, araD, epaE,
• (iv) phoA, araB, araC, araD, epaE, and
• (v) orfA, orfB, orfC, epaE, araB, and
• wherein
• the amino acid sequence of the protein encoded by the arab gene is SEQ ID NO:
• 58,
• the amino acid sequence of the protein encoded by the araC gene is SEQ ID NO:
• 60,
• the amino acid sequence of the protein encoded by the araD gene is SEQ ID NO:
• 62,
• the amino acid sequence of the protein encoded by the phoA gene is SEQ ID NO:
• 66,
• the amino acid sequence of the protein encoded by the phoB gene is SEQ ID NO:
• 68,
• the amino acid sequence of the protein encoded by the phoC gene is SEQ ID NO:
• 70,
• the amino acid sequence of the protein encoded by the phoD gene is SEQ ID NO:
• 72,
• the amino acid sequence of the protein encoded by the epaA gene is SEQ ID NO:
• 74,
• the amino acid sequence of the protein encoded by the epaE gene is SEQ ID NO:
• 82,
• the amino acid sequence of the protein encoded by the orfA gene is SEQ ID NO:
• 84,
• the amino acid sequence of the protein encoded by the orfB gene is SEQ ID NO:
• 86 or 90, and
• the amino acid sequence of the protein encoded by the orfC gene is SEQ ID NO:
• 88 or 92.