US 12,134,787 B2
Method for increasing the yield of oxidosqualene, triterpenes and/or triterpenoids and host cell therefore
Christian Schulze Gronover, Muenster (DE); Lowis Gerrit Boje Mueller, Rheine (DE); Nicole Van Deenen, Muenster (DE); and Jan Niklas Broeker, Muenster (DE)
Assigned to Fraunhofer-Gesellschaft zur Foerderung der Angewandten Forschung E.V., Munich (DE); and Westfaelische Wilhelms-Universitaet Muenster, Muenster (DE)
Appl. No. 17/046,207
Filed by Fraunhofer-Gesellschaft zur Foerderung der Angewandten Forschung E.V., Munich (DE); and Westfaelische Wilhelms-Universitaet Muenster, Muenster (DE)
PCT Filed Apr. 8, 2019, PCT No. PCT/EP2019/058789
§ 371(c)(1), (2) Date Oct. 8, 2020,
PCT Pub. No. WO2019/197327, PCT Pub. Date Oct. 17, 2019.
Claims priority of application No. 18166374 (EP), filed on Apr. 9, 2018.
Prior Publication US 2021/0071150 A1, Mar. 11, 2021
Int. Cl. C12N 9/04 (2006.01); C12N 1/16 (2006.01); C12N 1/18 (2006.01); C12N 9/90 (2006.01); C12N 15/81 (2006.01); C12P 5/00 (2006.01); C12P 33/00 (2006.01)
CPC C12N 9/0006 (2013.01) [C12N 1/16 (2013.01); C12N 1/18 (2013.01); C12N 9/90 (2013.01); C12N 15/81 (2013.01); C12P 5/007 (2013.01); C12P 33/00 (2013.01); C12Y 101/01034 (2013.01); C12Y 504/99007 (2013.01); C12Y 504/99041 (2013.01); C12N 2500/20 (2013.01)] 19 Claims
 
1. A recombinant host cell for manufacturing one or more of oxidosqualene(s), triterpene(s) and triterpenoid(s), wherein the host cell is engineered to overexpress a 3-hydroxy-3-methylglutaryl-coenzyme A reductase comprising the amino acid sequence as shown in SEQ ID NO: 1, and to overexpress a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 3, 4, 5, 6, 7 and 8, and wherein the one or more of oxidosqualene(s), triterpene(s) and triterpenoid(s) is/are selected from the group consisting of an oxidosqualene, a sterol, a pentacyclic triterpene, an acylated triterpene, β-amyrin, α-amyrin, taraxasterol, a triterpene acetate, an acylated triterpene, taraxerol, taraxerone, α-amyrone, β-amyrone, taraxasterone, friedelin, betulin, betulinic acid, cholesterol, ergosterol, lanosterol, a glucocorticoid, a mineralocorticoid, an estrogen, a gestagen, a cardenolide, a bufadienolide, and a steroidalkaloid,
wherein the host cell comprises a knock out of at least one locus selected from the group consisting of ROX1 (SEQ ID NO: 25), BTS1 (SEQ ID NO: 54), YPL062W (SEQ ID NO: 55), DOS2 (SEQ ID NO: 56), YER134C (SEQ ID NO: 57), VBA5 (SEQ ID NO: 58), YNR063W (SEQ ID NO: 59), YJL064W (SEQ ID NO: 60) and YGR259C (SEQ ID NO: 61).