Omega-hydroxylase-related fusion polypeptides with improved properties
The disclosure relates to omega-hydroxylase-related fusion polypeptides that result in improved omega-hydroxylated fatty acid derivative production when expressed in recombinant host cells. The disclosure further relates to microorganisms for expressing the omega-hydroxylase-related fusion polypeptides for the production of omega-hydroxylated fatty acid derivatives.
1. A CYP153A-reductase hybrid fusion polypeptide variant comprising at least 95% sequence identity to SEQ ID NO: 38 and having at least one mutation at an amino acid position corresponding to amino acid position 9, 10, 12, 13, 14, 28, 61, 119, 327, 413, 703, 745, 747, 749, 757, 770, 771 and 784.
2. The CYP153A-reductase hybrid fusion polypeptide variant of claim 1 , wherein the CYP153A-reductase hybrid fusion polypeptide variant is a hybrid cyp153ARedRhF fusion protein variant.
3. The CYP153A-reductase hybrid fusion polypeptide variant of claim 1 , wherein said mutation is selected from the group consisting of D9N, D9K, D10Y, Q12W, Q12R, Q12T, S13K, R14F, Q28M, Q28T, N61L, K119R, P327D, Y413R, L703G, P745R, D747N, E749L, E757A, T770G, V771F, and M784I.
4. The CYP153A-reductase hybrid fusion polypeptide variant of claim 3 , wherein expression of said CYP153A-reductase hybrid fusion polypeptide variant in a recombinant host cell results in a higher titer of an omega-hydroxylated fatty acid as compared to a titer of an omega-hydroxylated fatty acid produced by expression of a CYP153A-reductase hybrid fusion polypeptide of SEQ ID NO: 38.
5. The CYP153A-reductase hybrid fusion polypeptide variant of claim 3 , comprising any one of SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 74, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, and SEQ ID NO: 96.