	US 12,416,027 B2
	Squalene hopene cyclase (SHC) variants
Eric Eichhorn, Zürich (CH); and Christophe Ullmann, Nimes (FR)
Assigned to GIVAUDAN SA, Vernier (CH)
Appl. No. 17/776,686
Filed by Givaudan SA, Vernier (CH)
PCT Filed Dec. 3, 2020, PCT No. PCT/EP2020/084491 § 371(c)(1), (2) Date May 13, 2022, PCT Pub. No. WO2021/110848, PCT Pub. Date Jun. 10, 2021.
Claims priority of application No. 1917688 (GB), filed on Dec. 4, 2019.
Prior Publication US 2023/0021613 A1, Jan. 26, 2023
Int. Cl. C12P 17/02 (2006.01); C12N 9/90 (2006.01)

CPC C12P 17/02 (2013.01) [C12N 9/90 (2013.01); C12Y 504/99017 (2013.01)]

13 Claims

1. A process for preparing (−)-Ambrox or a mixture comprising (−)-Ambrox, the process comprising enzymatically converting (3E,7E)-homofarnesol (EEH) or a mixture of isomers of homofarnesol comprising EEH to (−)-Ambrox or a mixture comprising (−)-Ambrox using a squalene hopene cyclase/homofarnesol Ambrox cyclase (SHC/HAC) enzyme variant,

wherein the SHC/HAC enzyme variant has an amino acid sequence having at least 70.0% identity to SEQ ID NO: 1,

wherein the SHC/HAC enzyme variant amino acid sequence has amino acid alterations relative to SEQ ID NO: 1 at positions corresponding to positions 132, 224 and 432 of SEQ ID NO: 1 which are M132R, A224V and I432T respectively, and

wherein the SHC/HAC enzyme variant amino acid sequence has amino acid alterations relative to SEQ ID NO: 1 at a position corresponding to position 557 of SEQ ID NO: 1 and at least one position corresponding to position 81, 431 or 613 of SEQ ID NO: 1.