Patent ID: 7063956

Claim:
A process for the preparation of canthaxanthin wherein the process comprises culturing a recombinant cell containing farnesyl pyrophosphate and isopentyl pyrophosphate under culture conditions sufficient for the expression of enzymes which catalyze the conversion of the farnesyl pyrophosphate and isopentyl pyrophosphate to canthaxanthin, the recombinant cell being a host cell transformed by an expression vector comprising a regulatory sequence and a polynucleotide containing the following DNA sequences which encode the enzymes: a) a DNA sequence which encodes the geranylgeranyl pyrophosphate (GGPP) synthase of Flavobacterium sp. R1534 (crtE) (SEQ ID NO: 2) or a DNA sequence that hybridizes to a complementary strand of SEQ ID NO: 1 under the following conditions: hybridization in 7% sodium dodecyl sulfate (SDS), 1% bovine serum albumin (BSA), 0.5 M Na 2 HPO 4 , pH 7.2, at 65° C., washing twice for 5 minutes each in 2×SSC. 1% SDS, at room temperature, followed by two additional washes for 15 minutes each in 0.1% SSC, 0.1% SDS, at 65° C., wherein the hybrid DNA encodes a polypeptide having geranylgeranyl pyrophosphate (GGPP) synthase activity, b) a DNA sequence which encodes the prephytoene synthase of Flavobacterium sp. R1534 (crtB) (SEQ ID NO: 3) or a DNA sequence that hybridizes to SEQ ID NO: 1 under the following conditions: hybridization in 7% sodium dodecyl sulfate (SDS), 1% bovine serum albumin (BSA), 0.5 M Na 2 HPO 4 , pH 7.2, at 65° C., washing twice for 5 minutes each in 2×SSC, 1% SDS, at room temperature, followed by two additional washes for 15 minutes each in 0.1% SSC, 0.1% SDS, at 65° C., wherein the hybrid DNA encodes a polypeptide having prephytoene synthase activity, c) a DNA sequence which encodes the phytoene desaturase of Flavobacterium sp. R1534 (crtl) (SEQ ID NO: 4) or a DNA sequence that hybridizes to SEQ ID NO: 1 under the following conditions: hybridization in 7% sodium dodecyl sulfate (SDS). 1% bovine serum albumin (BSA), 0.5 M Na 2 HPO 4 , pH 7.2, at 65° C., washing twice for 5 minutes each in 2×SSC 1% SDS, at room temperature, followed by two additional washes for 15 minutes each in 0.1% SSC, 0.1% SDS, at 65° C., wherein the hybrid DNA encodes a polypeptide haying phytoene desaturase activity, d) a DNA sequence which encodes the lycopene cyclase of Flavobacterium sp. R1534 (crtY) (SEQ ID NO: 5) or a DNA sequence that hybridizes to SEQ ID NO: 1 under the following conditions: hybridization in 7% sodium dodecyl sulfate (SDS), 1% bovine serum albumin (BSA), 0.5 M Na 2 HPO 4 , pH 7.2, at 65° C., washing twice for 5 minutes each in 2×SSC, 1% SDS, at room temperature, followed by two additional washes for 15 minutes each in 0.1% SSC, 0.1% SDS, at 65° C., wherein the hybrid DNA encodes a polypeptide having lycopene cyclase activity, and e) a DNA sequence which encodes the β-carotene β4-oxygenase of microorganism E-396 (crtW E396 ) (SEQ ID NO: 32); and isolating the canthaxanthin from such cells or the culture medium.