# US 7,745,170 B2 · Process for producing carotenoid compound

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- Number: US 7,745,170 B2
- Document ID: US7745170B2
- Status: Granted
- Kind: B2
- Application number: 10/571,902
- Filed: 2004-09-08
- Published: 2007-05-10
- Granted: 2010-06-29
- Earliest claimed: 2003-09-17
- Assignees: Nippon Oil Co., Ltd.
- Inventors: Akira Tsubokura; Hisashi Yoneda; Kazuaki Hirasawa
- Web page: https://patentgrep.com/patent/US7745170B2

## CPC

- C12P 23/00: Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes (containing heterorings C12P17/00)
- A23L 5/44: using carotenoids or xanthophylls
- A23L 33/105: Plant extracts, their artificial duplicates or their derivatives
- C09B 61/00: Dyes of natural origin prepared from natural sources {, e.g. vegetable sources}
- C12N 15/01: Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor
- A23V 2002/00: Food compositions, function of food ingredients or processes for food or foodstuffs
- 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)
- C09B: ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES {, e.g. PIGMENTS}; MORDANTS; LAKES (fermentation or enzyme using processes to synthesise a desired chemical compound C12P)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A23V: INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION

## Abstract

The present invention relates to a process for producing zeaxanthin, β-carotene, or lycopene, comprising inducing mutation in a carotenoid-producing microorganism in which the base sequence of DNA corresponding to 16S ribosomal RNA is substantially homologous to the base sequence described in SEQ ID NO: 1; screening for a mutant strain having a high product proportion of zeaxanthin, β-carotene, or lycopene to the whole production amount of carotenoids to provide a microorganism producing zeaxanthin, β-carotene, or lycopene; culturing the mutant microorganism; and harvesting zeaxanthin, β-carotene, lycopene or a carotenoid mixture containing the same from the resultant culture.

## Claims

### Claim 1

A process for producing a carotenoid mixture containing zeaxanthin and β-cryptoxanthin, comprising
mutating a parent astaxanthin-producing bacterium having a 16S ribosomal RNA that is not less than 98% homologous to sequence of SEQ ID NO: 1;
culturing the mutants obtained to form colonies on a solid medium;
selecting yellow to orange colonies on said solid medium;
culturing the selected mutant colonies in a culture medium;
extracting and analyzing the carotenoids in the culture;
screening for a mutant strain having 20% or more proportion by mass of zeaxanthin to total carotenoids in said culture to provide a bacterium producing zeaxanthin and β-cryptoxanthin;
culturing said bacterium; and harvesting a carotenoid mixture containing zeaxanthin and β-cryptoxanthin from the resultant culture.

### Claim 2 (depends on 1)

The process according to claim 1, wherein the proportion of each of echinenone, 3-hydroxyechinenone, asteroidenone, canthaxanthin, adonirubin, adonixanthin, and astaxanthin produced by said zeaxanthin-producing microorganism is less than 10% proportion by mass to the total amount of carotenoids in the carotenoid mixture produced.

### Claim 3 (depends on 1)

The process according to claim 1, wherein said carotenoid mixture containing zeaxanthin further comprises β-carotene.

### Claim 4 (depends on 1)

The process according to claim 1, wherein said parent astaxanthin-producing bacterium is selected from the group consisting of strain E-396 (FERM BP-4283) and strain A-581-1 (FERM BP-4671).