ENGINEERED ALPHA-GUAIENE SYNTHASES
The current disclosure relates to compositions and methods relating to engineered polypeptides that can encode guaiene synthases that are selective for alpha-guaiene. The engineered polypeptides can also catalyze the production of increased amounts of alpha-guaiene. The disclosure also relates to host cells including the engineered polypeptides that can be used to produce large quantities of alpha-guaiene and its derivatives. The disclosure also relates to methods of producing alpha-guaiene employing said host cells.
1 . An engineered polypeptide encoding a guaiene synthase, wherein the engineered polypeptide comprises an amino acid sequence at least about 85% identical to SEQ ID NO:4 and wherein the amino acid sequence comprises a deletion of amino acids corresponding to positions 60-64 of SEQ ID NO:4.
2 - 40 . (canceled)
41 . The engineered polypeptide of claim 1 , wherein the engineered polypeptide comprises an amino acid sequence at least about 90% identical to SEQ ID NO:8.
42 . The engineered polypeptide of claim 1 , wherein the engineered polypeptide comprises an amino acid sequence at least about 95% identical to SEQ ID NO:8.
43 . The engineered polypeptide of claim 1 , wherein the engineered polypeptide comprises an amino acid sequence at least about 98% identical to SEQ ID NO:8.
44 . The engineered polypeptide of claim 1 , wherein the engineered polypeptide comprises an amino acid sequence that is SEQ ID NO:8.
45 . A cell expressing the engineered polypeptide of claim 1 .
46 . The cell of claim 45 , wherein the engineered polypeptide comprises an amino acid sequence at least about 90% identical to SEQ ID NO:8.
47 . The cell of claim 45 , wherein the engineered polypeptide comprises an amino acid sequence at least about 95% identical to SEQ ID NO:8.
48 . The cell of claim 45 , wherein the engineered polypeptide comprises an amino acid sequence at least about 98% identical to SEQ ID NO:8.
49 . The cell of claim 45 , wherein the engineered polypeptide comprises an amino acid sequence that is SEQ ID NO:8.
50 . The cell of claim 45 , wherein the cell is a microbial cell.
51 . The cell of claim 50 , wherein the microbial cell is yeast or E. coli.
52 . The cell of claim 51 , wherein the microbial cell is yeast.
53 . The cell of claim 52 , wherein the yeast is Saccharomyces cerevisiae.
54 . The cell of claim 45 , capable of producing total guaiene products comprising alpha-guaiene, wherein the percentage of the alpha-guaiene is at least greater than about 15% of the total guaiene products.
55 . The cell of claim 54 , wherein the percentage of alpha-guaiene is about 30% to about 90% of the total guaiene products.
56 . A nucleic acid molecule encoding the engineered polypeptide of claim 1 .
57 . The nucleic acid molecule of claim 56 , comprising a nucleic acid sequence of SEQ ID NO: 7.
58 . A method for producing guaiene, the method comprising:
growing the cell of claim 45 in a culture, and
collecting guaiene from the culture.