	US 12,404,310 B2
	Maltose dependent degrons, maltose-responsive promoters, stabilization constructs, and their use in production of non-catabolic compounds
Penelope R. Chua, Emeryville, CA (US); Hanxiao Jiang, Emeryville, CA (US); and Adam Leon Meadows, Emeryville, CA (US)
Assigned to AMYRIS, INC., Emeryville, CA (US)
Filed by AMYRIS, INC., Emeryville, CA (US)
Filed on Sep. 4, 2020, as Appl. No. 17/013,246.
Application 17/013,246 is a division of application No. 15/738,555, granted, now 10,808,015, previously published as PCT/US2016/039386, filed on Jun. 24, 2016.
Claims priority of provisional application 62/184,793, filed on Jun. 25, 2015.
Claims priority of provisional application 62/266,436, filed on Dec. 11, 2015.
Prior Publication US 2020/0399326 A1, Dec. 24, 2020
Int. Cl. C07K 14/395 (2006.01); C12N 1/16 (2006.01); C12N 15/52 (2006.01); C12N 15/63 (2006.01); C12N 15/65 (2006.01); C12P 5/02 (2006.01); C12P 7/64 (2022.01); C12P 19/62 (2006.01); C12P 21/00 (2006.01); C12P 21/02 (2006.01)

CPC C07K 14/395 (2013.01) [C12N 1/16 (2013.01); C12N 15/52 (2013.01); C12N 15/635 (2013.01); C12N 15/65 (2013.01); C12P 5/02 (2013.01); C12P 5/026 (2013.01); C12P 7/64 (2013.01); C12P 19/62 (2013.01); C12P 21/00 (2013.01); C12P 21/02 (2013.01); C07K 2319/95 (2013.01); C12N 2330/51 (2013.01); C12Y 503/03002 (2013.01)]

12 Claims

1. A genetically modified host cell comprising:

a heterologous nucleic acid encoding a fusion protein comprising a protein of interest fused in frame to a maltose dependent degron,

wherein the fusion protein is most stable when the maltose dependent degron is in contact with a maltose based inducer compared to when the maltose dependent degron is not in contact with the maltose based inducer,

wherein the maltose degron in the fusion protein comprises:

i. the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 28;

ii. amino acid residues 1-365 of the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 28; or

iii. a homolog comprising at least 90% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 28; and

wherein the maltose dependent degron further comprises one or more variant amino acid residue substitutions selected from the group consisting of K7R, I10T, W11G, L21S, V24A, F28Y, D42V, K43E, A64T, F68S, D83G, D88N, P92T, W95R, V98I, N101I, A110T, I117V, P134S, A135T, L136M, M149I, Y168C, Y168N, Y177H, N186S, A187P, L193S, D198V, D210E, A216V, A217D, G229C, I236N, D237N, N242D, L263M, L291V, A304S, T321N, M322L, A339T, A351T, T357S, T367S, S370P, and N374S, wherein said one or more variant amino acid residue substitutions correspond to the amino acid positions of the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 28.