DNA strands useful for the synthesis of xanthophylls and the pro

Details DNA strands useful for the synthesis of xanthophylls and the pro DNA strands useful for the synthesis of xanthophylls and the pro

1996-09-23

1998-09-22

Wax, Robert A.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing oxygen-containing organic compound

435189, 4352521, 4352523, 43525233, 435 67, 4352551, 4353201, 536 231, 536 232, 536 237, C12P 726

Patent

active

058112731

DESCRIPTION:

BRIEF SUMMARY
TECHNICAL FIELD

The present invention relates to DNA strands useful for the synthesis of keto group-containing xanthophylls (ketocarotenoids) such as astaxanthin which are useful for heightening the color of cultured fishes and shellfishes such as sea breams, salmons, lobster and the like and is used for foods as a coloring agent and an antioxidant, and to a process for producing keto group-containing xanthophylls (ketocarotinoids) such as astaxanthin with use of a microorganism into which the DNA strands have been introduced.


BACKGROUND ART

The term xanthophylls mean carotenoid pigments having an oxygen-containing group such as a hydroxyl group, a keto group or an epoxy group. Carotenoids are synthesized by the isoprenoid biosynthetic process which is used in common halfway with steroids and other terpenoids with mevalonic acid as a starting material. C15 farnesyl pyrophosphate (FPP) resulting from isoprene basic biosynthetic pathway is condensed with C5 isopentenyl pyrophosphate (IPP) to give C20 geranylgeranyl pyrophosphate (GGPP). Two molecules of GGPP are condensed to synthesize a colorless phytoene as an initial carotenoid. The phytoene is converted into phytofluene, .zeta.-carotene, neurosporene and then lycopene by a series of desaturation reactions, and lycopene is in turn converted into .beta.-carotene by the cyclization reaction. It is believed that a variety of xanthophylls are synthesized by introducing a hydroxyl group or a keto group into the .beta.-carotene (See Britton, G., "Biosynthesis of Carotenoids"; Plant Pigments, Goodwin, T. W. ed., London, Academic Press, 1988, pp. 133-182).
The present inventors have recently made it possible to clone a carotenoid biosynthesis gene cluster from a epiphytic non-photosynthetic bacterium Erwinia uredovora in Escherichia coli with an index of the yellow tone of the bacterium, a variety of combinations of the genes being expressed in microorganisms such as Escherichia coli to produce phytoene, lycopene, .beta.-carotene, and zeaxanthin which is a derivative of .beta.-carotene into which hydroxyl groups have been introduced (See FIG. 10; Misawa, N., Nakagawa, M., Kobayashi, K., Yamano, S., Izawa, Y., Nakamura, K., Harashima, K.; "Elucidation of the Erwinia uredovora Carotenoid biosynthetic Pathway by Functional Analysis of Gene Products Expressed in Escherichia coli", J. Bacteriol., 172, p.6704-6712, 1990; Misawa, N., Yamano, S., Ikenaga, H., "Production of .beta.-carotene in Zymomonas mobilis and Agrobacterium tumefaciencs by Introduction of the Biosynthesis Genes from Erwinia uredovora", Appl. environ. Microbiol., 57, p. 1847-1849, 1991; and Japanese Patent Application No. 58786/1991 (Japanese Patent Application No. 53255/1990): "DNA Strands useful for the Synthesis of Carotenoids").
On the other hand, astaxanthin, a red xanthophyll, is a typical animal carotenoid which occurs particularly in a wide variety of marine animals including red fishes such as a sea bream and a salmon, and crustaceans such as a crab and a lobster. In general, animals cannot biosynthesize carotenoids, so that it is necessary for them to ingest carotenoids synthesized by microorganisms or plants from their environments. Thus, astaxanthin has hitherto been used widely for strengthening the color of cultured fishes and shellfishes such as a sea bream, a salmon, a lobster and the like. Moreover, astaxanthin has attracted attention not only as a coloring matter in foods but also as an anti-oxidant for removing active oxygen generated in bodies, which causes carcinoma (see Takao Matsuno ed., "Physiological Functions and Bioactivities of Carotenoids in Animals", Kagaku to Seibutsu, 28, p. 219-227, 1990). As the sources of astaxanthin, there have been known crustaceans such as a krill in the Antarctic Ocean, cultured products of a yeast Phaffia, cultured products of a green alga Haematococcus, and products obtained by the organic synthetic methods. However, when crustaceans such as a krill in the Antarctic Ocean or the like are used, it requires laborious works and much expenses f

REFERENCES:
Yokoyama, et al. "Production of Astaxanthin and 4-Ketozeaxanthin by the Marine Bacterium, Agro-bacterium aurantiacum," Biosci. Biotech. Biochem. 58(10): 1842-44 (1994).
Misawa, et al. "Elucidation of the Erwinia unredovora Carotenoid Biosynthetic Pathway by Functional Analysis of Gene Products Expressed in Escherichia coli," J. Bacteriology 172(12): 6704-12 (Dec. 1990).
Misawa, et al. "Production of .beta.-Carotene in Zymomonas mobilis and Agrobacterium tumefaciens by Introduction of the Biosynthesis Genes from Erwinia uredovora," Applied and Environmental Microbiology 57(6): 1847-49 (Jun. 1991).
Misawa et al. "Structure and functional Analysis of a marine bacterial carotenoid biosynthesis gene cluster and astaxanthin biosynthetic pathway proposed at the gene level" J. Bacteriol. 177 (22), 6575-6584, Nov. 1995.

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