US 11,920,177 B2
Synthesis of beta-hydroxyisovalerate and methods of use
Goutham Vemuri, Maple Grove, MN (US); Christopher Lindsay, St. Paul, MN (US); Kevin Roberg-Perez, Minneapolis, MN (US); Christopher D. Snow, St. Paul, MN (US); and Elizabeth A. Cameron, New Brighton, MN (US)
Assigned to SASYA INC., St. Paul, MN (US)
Filed by Sasya Inc., St. Paul, MN (US)
Filed on Apr. 6, 2022, as Appl. No. 17/658,217.
Claims priority of provisional application 63/171,418, filed on Apr. 6, 2021.
Prior Publication US 2022/0315954 A1, Oct. 6, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 7/16 (2006.01); C12N 1/20 (2006.01); C12N 9/04 (2006.01); C12N 9/88 (2006.01); C12N 15/52 (2006.01)
CPC C12P 7/16 (2013.01) [C12N 1/20 (2013.01); C12N 9/0006 (2013.01); C12N 9/88 (2013.01); C12N 15/52 (2013.01); C12Y 101/01086 (2013.01)] 23 Claims
OG exemplary drawing
 
1. A non-natural microorganism capable of producing beta-hydroxyisovalerate (βHIV), wherein the non-natural microorganism comprises a non-natural enzyme, wherein the non-natural enzyme has been modified or mutated to inncrease the ability of the enzyme to preferentially utilize α-ketoisocaproate as its substrate, and wherein the non-natural enzyme comprises one or more modifications or mutations at substrate-specificity positions corresponding to amino acids selected from N187, W210, V212, V217, L224, S226, I227, V228, P239, N241, Q251, I252, Q265, L289, L323, F336, F347, A361, G362, N363, F364, L367, F368, and F371, of any of SEQ ID NO: 1 or SEQ ID NO: 6, and wherein the non-natural microorganism has been modified or mutated to provide more βHIV than its wild-type parent.