Microbial production of artemisinic acid and derivatives

An ADS variant comprising an amino acid substitution at one or more positions corresponding to the following positions of SEQ ID NO:

• 1:
• 396, 104, 162, 474, 118, 363, 322, 173, 112, 431, 151, 291, 134, 341, 230, 245, 44, 385, 100, 469, 500, 292, 471, 207, 463, 189, 340, 510, 260, 247, 211, 430, 277, 318, 275, 170, 124, 125, 145, 169, 445, 155, 152, 507, 520, 393, 447, 455, 498, 409, 204, and 261 wherein the ADS variant has increased amorphadiene synthase activity as compared to SEQ ID NO:
• 1.

An AO variant comprising an amino acid substitution at one or more positions corresponding to the following positions of SEQ ID NO:

• 3:
• 239, 257, 408, 410, 421, 320, 130, 489, 198, 119, 102, 186, 252, 294, 314, 457, 474, 319, 322, 390, 125, 155, 251, 445, 424, 250, 387, 462, 153, 151, 243, 308, 495, 73, 103, 59, 123, 124, 146, 256, 261, 369, 469, and 64 wherein the AO variant has increased amorphadiene oxidase activity as compared to SEQ ID NO:
• 3.

An ADH variant comprising an amino acid substitution at one or more positions corresponding to the following positions of SEQ ID NO:

• 6:
• 82, 302, 155, 360, 299, 258, 304, 19, 107, 193, 263, 168, 78, 20, 253, 75, 191, 302, 80, 153, 203, 169, 229, 221, 329, 150, 6, 305, 60, 25, 310, 92, 233, 257, and 170 wherein the ADH variant has increased alcohol dehydrogenase activity.

A method for producing artemisinin, comprising:

• contacting one or more enzymes comprising an amino acid sequence having at least 70%, or at least about 80%, at least about 90%, or at least about 95% sequence identity to one of SEQ ID NOs:
• 11 to 488, with a substrate selected from one or more of artemisinic acid (AA) and dihydroartemisinic acid (DHAA), and
• recovering artemisinin.