Fermentative production of n-butylacrylate using alcohol acyl transferase enzymes
AssigneeBASF SE
InventorsStephan Saum, Woncheol Kim, Oskar Zelder, Jennifer Jaitzig, Zheyuan Guo
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Abstract
Provided herein is a recombinant nucleic acid molecule, a recombinant microorganism, and a method for fermentative production of n-butylacrylate and other esters from alcohols and acyl-CoA units using alcohol acyl transferase enzymes.
Claims9
A recombinant microorganism comprising a nucleic acid molecule encoding an alcohol acyl transferase (AAT) gene encoding an AAT enzyme having an n-butylacrylate (n-BA) forming activity, wherein the microorganism also comprises a butanol producing pathway and an acryloyl-CoA producing pathway, and wherein the microorganism has an introduced, increased, or enhanced n-butylacrylate forming activity and/or expression of said AAT enzyme;
- wherein the AAT gene encoding an AAT enzyme having an n-BA forming activity is selected from the group consisting of:
- (I) a nucleic acid molecule comprising a sequence selected from the group consisting of SEQ ID NO:
- 1, 3, 5, 7, 9, 11, and 13;
- (II) a nucleic acid molecule having at least 95% identity to a nucleic acid molecule selected from the group consisting of SEQ ID NO:
- 1, 3, 5, 7, 9, 11, and 13;
- (III) a nucleic acid molecule hybridizing to the full complement of a nucleic acid molecule having a sequence selected from the group consisting of SEQ ID NO:
- 1, 5, 7, 9, 11, and 13;
- (IV) a nucleic acid molecule encoding a polypeptide selected from the group consisting of SEQ ID NO:
- 2, 6, 8, 10, 12, and 14;
- and
- (V) a nucleic acid molecule encoding a polypeptide having at least 95% identity to a polypeptide selected from the group consisting of SEQ ID NO:
- 2, 6, 8, 10, 12, and 14.
A method for producing a recombinant microorganism producing n-BA comprising the steps of:
- (I) introducing, increasing, or enhancing the activity and/or expression of an alcohol acyl transferase (AAT) gene encoding an AAT enzyme having an n-BA forming activity in a microorganism;
- and
- (II) further introducing in the microorganism a butanol producing pathway and an acryloyl-CoA producing pathway;
- wherein the AAT gene encoding an AAT enzyme having an n-BA forming activity is selected from the group consisting of:
- (I) a nucleic acid molecule comprising a sequence selected from the group consisting of SEQ ID NO:
- 1, 3, 5, 7, 9, 11, and 13;
- (II) a nucleic acid molecule having at least 95% identity to a nucleic acid molecule selected from the group consisting of SEQ ID NO:
- 1, 3, 5, 7, 9, 11, and 13;
- (III) a nucleic acid molecule hybridizing to the full complement of a nucleic acid molecule having a sequence selected from the group consisting of SEQ ID NO:
- 1, 5, 7, 9, 11, and 13;
- (IV) a nucleic acid molecule encoding a polypeptide selected from the group consisting of SEQ ID NO:
- 2, 6, 8, 10, 12, and 14;
- and
- (V) a nucleic acid molecule encoding a polypeptide having at least 95% identity to a polypeptide selected from the group consisting of SEQ ID NO:
- 2, 6, 8, 10, 12, and 14.
A recombinant expression construct comprising a promoter functional in a microorganism functionally linked to a nucleic acid molecule having a sequence selected from the group consisting of:
- (I) a nucleic acid molecule comprising a sequence selected from the group consisting of SEQ ID NO:
- 1, 3, 5, 7, 9, 11, and 13;
- and
- (II) a nucleic acid molecule having at least 95% identity to a nucleic acid molecule selected from the group consisting of SEQ ID NO:
- 1, 3, 5, 7, 9, 11, and 13;
- and
- (III) a nucleic acid molecule hybridizing to the full complement of a nucleic acid molecule having a sequence selected from the group consisting of SEQ ID NO:
- 1, 5, 7, 9, 11, and 13;
- and
- (IV) a nucleic acid molecule encoding a polypeptide of SEQ ID NO:
- 2, 6, 8, 10, 12, and 14;
- and
- (V) a nucleic acid molecule encoding a polypeptide having at least 95% identity to a polypeptide selected from the group consisting of SEQ ID NO:
- 2, 6, 8, 10, 12, and 14,
- wherein the promoter is heterologous to the nucleic acid molecule.
Dependencies are inferred from the claim text.