Acyl-ACP reductase with improved properties
The disclosure relates to acyl-ACP reductase (AAR) enzyme variants that result in improved fatty aldehyde and fatty alcohol production when expressed in recombinant host cells. The disclosure further relates to methods of making and using such AAR variants for the production of fatty alcohol compositions having particular characteristics.
1. A variant acyl-ACP reductase (AAR) polypeptide comprising at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 79, wherein said variant AAR polypeptide comprises a tryptophan at amino acid position 18, and wherein said AAR polypeptide catalyzes the conversion of an acyl-ACP to a fatty aldehyde.
2. The variant AAR polypeptide of claim 1 , wherein expression of said variant AAR polypeptide in a recombinant host cell results in a higher titer of a fatty alcohol composition as compared to a titer produced by expression of a wild type AAR polypeptide in a corresponding wild type host cell.
3. The variant AAR polypeptide of claim 2 , wherein said fatty alcohol composition is a C12 or C14 fatty alcohol composition.
4. The variant AAR polypeptide of claim 1 , wherein said variant AAR polypeptide further comprises a mutation at an amino acid position selected from the group consisting of amino acids 8, 16, 21, 24, 31, 34, 35, 43, 50, 63, 86, 112, 113, 116, 118, 120, 135, 148, 153, 154, 155, 157, 159, 168, 172, 187, 188, 191, 209, 210, 211, 236, 277, 281, 283, 285, 291, 324, 328, 335, 337 and 338.
5. The variant AAR polypeptide of claim 4 , wherein said mutation is selected from the group consisting of L8A, D16L, M21L, D24E, D24Y, D24V, D24P, L31V, L31M, W34F, W35F, D43E, A50Q, C63A, C63G, C63Y, S86G, A112R, S113K, Q116G, R118Q, T1205, A1355, T148C, T148E, T148V, I153P, T154A, Q155C, Q155L, T157V, A159V, I168V, C172L, T187V, T188H, T188V, Q191A, L209R, E210Y, A211W, T236C, Q277V, A281L, E283G, E283S, A285V, M291V, A324T, A328S, Q335N, L337V and L338W.
6. The variant AAR polypeptide of claim 5 , wherein said variant AAR polypeptide comprises a M21L mutation, a C63GA mutation, an S113K mutation, an T154A mutation, an A281L mutation, or an L8A mutation.
7. The variant AAR polypeptide of claim 1 , wherein said variant AAR polypeptide further comprises
(a) a M21L mutation, a C63G mutation, a S113K mutation, a T154A mutation, and an A281L mutation;
(b) a L8A mutation, a M21L mutation, a C63G mutation, a S113K mutation, a T154A mutation, and an A281L mutation;
(c) a D16L mutation, a M21L mutation, a C63G mutation, a S113K mutation, a T154A mutation, and an A281L mutation;
(d) a L8A mutation, a D24V mutation, a C63G mutation, a S113K mutation, a Q155L mutation, and an A281L mutation;
(e) a D24P mutation, a L31M mutation, a C63G mutation, a S113K mutation, a T154A mutation, and an A281L mutation;
(f) L8A mutation, a D16L mutation, a D24V mutation, a C63G mutation, a S113K mutation, a T154A mutation, and an A281L mutation; or
(g) a D24E mutation, a C63G mutation, a S113K mutation, a T154A mutation, and an A281L mutation.