	US 12,234,495 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. 17/658,214.
Claims priority of provisional application 63/171,418, filed on Apr. 6, 2021.
Prior Publication US 2022/0325304 A1, Oct. 13, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 7/16 (2006.01); C12N 1/20 (2006.01); C12N 1/21 (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)]

20 Claims

1. A non-natural enzyme capable of producing beta-hydroxyisovalerate (βHIV), the non-natural enzyme comprising one or more amino acid modifications or mutations relative to a corresponding enzyme lacking the substitutions, wherein the non-natural enzyme comprises one or more modifications or mutations at substrate-specificity positions corresponding to amino acids selected from V212, L224, Q251 and F371, of any of SEQ ID NO: 1 or SEQ ID NO: 6, and wherein the non-natural enzyme is modified or mutated to provide more beta-hydroxyisovalerate (βHIV) synthase activity than a corresponding enzyme with the same amino acid sequence lacking the one or more amino acid modifications or mutations.