US 12,480,146 B2 · Microbial production of mogrol and mogrosides

Abstract

The present invention provides host cells and methods for making mogrol glycosides, including Mogroside V (Mog.V), Mogroside VI (Mog.VI), Iso-Mogroside V (Isomog.V), siamenoside, and glycosylation products that are minor products in Siraitia grosvenorii. The invention provides engineered enzymes and engineered host cells for producing mogrol glycosylation products, such as Mog.V, Mog.VI, and Isomog.V, at high purity and/or yield. The present technology further provides methods of making products containing mogrol glycosides, such as Mog.V, Mog.VI, and Isomog.V, including food products, beverages, oral care products, sweeteners, and flavoring products.

Claims20

Dependent claims

A method for making mogrol or mogroside, comprising:

providing a recombinant microbial host cell expressing a heterologous enzyme pathway catalyzing the conversion of isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) to mogrol or mogroside, the heterologous enzyme pathway comprising:
a farnesyl diphosphate synthase (FPPS),
a squalene synthase (SQS),
a squalene epoxidase (SQE) catalyzing the formation of 2,3;22,23-dioxidosqualene from squalene,
a triterpene cyclase (TTC) catalyzing the formation of 24,25-epoxycucurbitadienol from 2,3;22,23-dioxidosqualene,
an epoxide hydrolase (EPH) catalyzing the formation of 24,25-dihydroxycucurbitadienol from 24,25-epoxycucurbitadienol, and
a cytochrome P450 enzyme catalyzing the formation of mogrol from 24,25-dihydroxycucurbitadienol;
wherein the cytochrome P450 enzyme comprises an amino acid sequence having at least 85% sequence identity with SEQ ID NO:
171;
and
culturing the host cell under conditions for producing the mogrol or mogroside.

8 more elements

Dependencies are inferred from the claim text.

Facts

Application

17/785,488Filed 16 Dec 2020 · us provisional application

Priority

16 Dec 2019

Published

US 2023/0042171 A19 Feb 2023

Granted

25 Nov 2025

Term ends

Not estimatedThe source records don’t support an estimate.

Classification22 codes

C07K 14/415
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof: from plants
C12P 17/02
Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms: oxygen as only ring hetero atoms
C12N 9/0077
Acting on paired donors with incorporation of molecular oxygen: with a reduced iron-sulfur protein as one donor
C12N 9/0083
Acting on paired donors with incorporation of molecular oxygen: miscellaneous

Family

Application62/948,657Us provisional application · filed 16 Dec 2019

Application63/075,631Us provisional application · filed 8 Sep 2020

Application63/085,557Us provisional application · filed 30 Sep 2020

US 2023/0042171 A1Pre-grant publication · published 9 Feb 2023

US 12,480,146 B2This patent

From the US filing history. Foreign family members aren’t listed.

Microbial production of mogrol and mogrosides

Life of this patent

Microbial production of mogrol and mogrosides

Life of this patent

Abstract

Claims20