Patent ID: 6613543
Filing Date: 2003-09-02
Classification: C07K,C12N,C12P

Abstract:
A process for producing zeaxanthin comprising:a) culturing, in a suitable medium containing a starting material selected from the group consisting of farnesyl pyrophosphate, isopentyl pyrophosphate, geranylgeranyl pyrophosphate, prephytoene, phytoene, lycopene, &bgr;-carotene and combinations thereof, a transformed host cell containing an expression cassette comprising a polynucleotide having the following DNA sequences: i) a DNA sequence which encodes the geranygeranyl pyrophosphate (GGPP) synthase of Flavobacterium sp. R1534 (crtE) or a DNA sequence which specifically hybridizes with a complementary strand of the DNA sequence which encodes the GGPP synthase of Flavobacterium sp. R1534 (crtE) under conditions of hybridization utilizing 7% sodium dodecyl sulfate (SDS), 1% bovine serum albumin (BSA), 0.5 M Na2HPO4 pH 7.2 at 65Â° C., and encodes a protein having GGPP synthase activity, ii) a DNA sequence which encodes the prephytoene synthase of Flavobacterium sp. R1534 (crtB) or a DNA sequence which specifically hybridizes with a complementary strand of the DNA sequence which encodes the prephytoene synthase of Flavobacterium sp. R1534 (crtB) under conditions of hybridization utilizing 7% sodium dodecyl sulfate (SDS), 1% bovine serum albumin (BSA), 0.5 M Na2HPO4 pH 7.2 at 65Â° C., and encodes a protein having prephytoene synthase activity, iii) a DNA sequence which encodes the phytoene desaturase of Flavobacterium sp. R1534 (crtI) or a DNA sequence which specifically hybridizes with a complementary strand of the DNA sequence which encodes the phytoene desaturase of Flavobacterium sp. R1534 (crtI) under conditions of hybridization utilizing 7% sodium dodecyl sulfate (SDS), 1% bovine serum albumin (BSA), 0.5 M Na2HPO4 pH 7.2 at 65Â° C., and encodes a protein having phytoene desaturase activity, iv) a DNA sequence which encodes the lycopene cyclase of Flavobacterium sp. R1534 (crtY) or a DNA sequence which specifically hybridizes with a complementary strand of the DNA sequence which encodes the lycopene cyclase of Flavobacterium sp. R1534 (crtY) under conditions of hybridization utilizing 7% sodium dodecyl sulfate (SDS), 1% bovine serum albumin (BSA), 0.5 M Na2HPO4 pH 7.2 at 65Â° C., and encodes a protein having lycopene cyclase activity, and v) a DNA sequence which encodes the &bgr;-carotene hydroxylase of Flavobacterium sp. R1534 (crtZ) or a DNA sequence which specifically hybridizes with a complementary strand of the DNA sequence which encodes the &bgr;-carotene hydroxylase of Flavobacterium sp. R1534 (crtZ) under conditions of hybridization utilizing 7% sodium dodecyl sulfate (SDS), 1% bovine serum albumin (BSA), 0.5 M Na2HPO4 pH 7.2 at 65Â° C., and encodes a protein having &bgr;-carotene hydroxylase activity, the polynucleotide being substantially free of other polynucleotides of Flavobacterium sp. R1534; andb) isolating zeaxanthin from such cells or the culture medium.