# US 9,752,174 B2 · High-purity steviol glycosides

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- Number: US 9,752,174 B2
- Document ID: US9752174B2
- Status: Granted
- Kind: B2
- Application number: 14/287,837
- Filed: 2014-05-27
- Published: 2014-12-04
- Granted: 2017-09-05
- Earliest claimed: 2013-05-28
- Assignees: PureCircle Sdn Bhd; The Coca-Cola Company
- Applicants: Purecircle Sdn Bhd; The Coca-Cola Company
- Inventors: Avetik Markosyan; Indra Prakash; Cynthia Bunders; Pankaj Soni; Jarrin Cyrille; Aurélien Badie; Robert ter Halle
- Web page: https://patentgrep.com/patent/US9752174B2

## CPC

- C12P 19/56: having an oxygen atom of the saccharide radical directly bound to a condensed ring system having three or more carbocyclic rings, e.g. daunomycin, adriamycin
- A23L 2/60: Sweeteners
- A23L 27/33: {containing sugars or derivatives}
- A24B 15/10: Chemical features of tobacco products or tobacco substitutes
- A24B 15/302: {by natural substances obtained from animals or plants}
- C07H 15/256: Polyterpene radicals
- A23V 2002/00: Food compositions, function of food ingredients or processes for food or foodstuffs
- Y02P 20/582: Recycling of unreacted starting or intermediate materials
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B)
- A24B: MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- A23V: INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

Methods of preparing highly purified steviol glycosides, particularly rebaudiosides A, D and M are described. The methods include utilizing recombinant microorganisms for converting various staring compositions to target steviol glycosides. In addition, novel steviol glycosides reb D2 and reb M2 are disclosed, as are methods of preparing the same. The highly purified rebaudiosides are useful as non-caloric sweetener in edible and chewable compositions such as any beverages, confectioneries, bakery products, cookies, and chewing gums.

## Claims

### Claim 1

A method for preparing reb D2 comprising:
a. contacting a starting composition comprising reb A with UDP-glucosyltransferase, to produce a composition comprising reb D2; and
b. isolating the composition comprising reb D2.

### Claim 2 (depends on 1)

The method of claim 1, further comprising purifying the composition comprising reb D2 to provide reb D2 having a purity greater than about 95% by weight on an anhydrous basis.

### Claim 3

A method for preparing reb M2 comprising:
a. contacting a starting composition comprising reb D2 with UDP-glucosyltransferase to produce a composition comprising reb M2; and
b. isolating the composition comprising reb M2.

### Claim 4 (depends on 3)

The method of claim 3, further comprising purifying the composition comprising reb M2 to provide reb M2 having a purity greater than about 95% by weight on an anhydrous basis.