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AssigneeGenomatica, Inc.
InventorsAndreas W. SCHIRMER, Erin Frances PERRY, Alma Itzel RAMOS-SOLIS, Angelica ZABALA-BAUTISTA
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Recombinant proteobacteria, including γ-proteobacteria, comprising a heterologous acyl-ACP desaturase and a heterologous acyl-ACP thioesterase, wherein the native dual 3-hydroxy acyl-ACP dehydratase/isomerase is deleted are provided herein. The recombinant proteobacteria produce non-native monounsaturated free fatty acids or derivatives thereof. Methods of producing non-native monounsaturated free fatty acids or derivatives thereof are also provided, in addition to cell cultures and fatty acid compositions produced by the recombinant proteobacteria. The recombinant proteobacteria may be used to produce insect pheromones or precursors thereof, and fragrances or precursors thereof.
A recombinant proteobacterium comprising a heterologous acyl-ACP desaturase and a heterologous acyl-ACP thioesterase;
The recombinant proteobacterium of any one of the preceding claims, wherein the heterologous acyl-ACP-desaturase is Δ9-tetradecanoyl-acyl-ACP desaturase.
The recombinant proteobacterium of any one of the preceding claims, wherein the heterologous acyl-ACP desaturase has about 85% sequence identity to SEQ ID NO:
The recombinant proteobacteria of any one of the preceding claims, wherein the heterologous acyl-ACP thioesterase is a plant FatA-type thioesterase, a plant FatB-type thioesterase, or a bacterial acyl-ACP thioestersase.
The recombinant proteobacterium of any one of the preceding claims, wherein the heterologous acyl-ACP thioesterase has about 85% sequence identity to SEQ ID NO:
The recombinant proteobacterium of any one of the preceding claims, wherein the non-native monounsaturated free fatty acid is a ω5-monounsaturated free fatty acid or a derivative thereof.
The recombinant proteobacterium of any one of the preceding claims, wherein an amount of non-native monounsaturated free fatty acids or derivatives thereof produced by the recombinant proteobacterium is greater than 90% of the total amount of native and non-native monounsaturated free fatty acids or derivatives thereof produced by the recombinant proteobacterium.
The recombinant proteobacterium of any one of the preceding claims, wherein the non-native monounsaturated free fatty acid is selected from the group consisting of z9-tetradecenoic acid, z11-hexadecenoic acid, z13-octadecenoic acid, and a combination thereof.
The recombinant proteobacterium of any one of the preceding claims, further comprising a heterologous ferredoxin.
The recombinant proteobacterium of any one of the preceding claims, wherein the recombinant bacterium overexpresses endogenous flavodoxin reductase or ferredoxin reductase or expresses heterologous ferredoxin reductase or heterologous flavodoxin reductase.
The recombinant proteobacterium of any one of the preceding claims, wherein the recombinant proteobacterium further comprises a heterologous 3-hydroxy acyl-ACP-dehydratase (FabZ).
The recombinant proteobacterium of any one of the preceding claims, wherein the recombinant proteobacterium further comprises (1) an increased level of expression of FadR polypeptide, and/or (2) an increased level of DNA binding activity, compared to a corresponding wild-type proteobacterium.
The recombinant proteobacterium of any one of the preceding claims, wherein the recombinant proteobacterium further comprises a heterologous carboxylic acid reductase.
The recombinant proteobacterium of any one of the preceding claims, wherein the recombinant bacterium further comprises a heterologous alcohol dehydrogenase.
The recombinant proteobacterium of any one of the preceding claims, wherein the recombinant bacterium further comprises a heterologous ω-hydroxylase.
The recombinant proteobacterium of any one of the preceding claims, wherein the fatty acid derivative comprises a fatty alcohol, an ω-hydroxy fatty acid, a fatty alcohol acetate, a fatty aldehyde, or an α/ω-dicarboxylic acid.
A nucleotide sequence encoding a Δ9-tetradecanoyl-acyl-ACP desaturase having at least 85% sequence identity to SEQ ID NO:
A vector comprising a nucleotide sequence encoding a Δ9-tetradecanoyl-acyl-ACP desaturase having at least 85% sequence identity to SEQ ID NO:
A recombinant host cell comprising a nucleotide sequence encoding a Δ9-tetradecanoyl-acyl-ACP desaturase having at least 85% sequence identity to SEQ ID NO:
A recombinant proteobacterium comprising a heterologous acyl-ACP desaturase, a heterologous ferredoxin, a heterologous flavodoxin reductase or a heterologous ferredoxin reductase, a heterologous carboxylic acid reductase, and a heterologous 3-hydroxy-acyl-ACP dehydratase;
A recombinant proteobacterium comprising a heterologous acyl-ACP desaturase, a heterologous ferredoxin, a heterologous flavodoxin reductase or a heterologous ferredoxin reductase, a heterologous acyl-ACP reductase and a heterologous alcohol dehydrogenase;
A recombinant proteobacterium comprising a heterologous acyl-ACP desaturase, a heterologous ferredoxin, a heterologous flavodoxin reductase or a heterologous ferredoxin reductase, a heterologous ω-hydroxylase, and a heterologous 3-hydroxy-acyl-ACP dehydratase;
Dependencies are inferred from the claim text.