Abstract: Disclosed are the following DNA strands relating to the synthesis of keto group-containing xanthophylls such as astaxanthin and the like, and the techniques relating to the production of xanthophylls by genetic engineering:A DNA strand having a nucleotide sequence which encodes a polypeptide having an enzyme activity for converting a methylene group at the 4-position of a .beta.-ionone ring into a keto group.A DNA strand having a nucleotide sequence which encodes a polypeptide having an enzyme activity for converting a methylene group at the 4-position of a 3-hydroxy-.beta.-ionone ring into a keto group.A DNA strand having a nucleotide sequence which encodes a polypeptide having an enzyme activity for adding a hydroxyl group to the 3-carbon of a 4-keto-.beta.-ionone ring.It is possible to produce a variety of xanthophylls such as canthaxanthin, astaxanthin and the like by introducing the DNA strands into an appropriate microorganism such as Escherichia coli and the like.

Abstract: Novel proteins of Flavobacrerium sp. R1534 and the DNA sequences which encode these proteins are disclosed which provide an improved biosynthetic pathway from farnesyl pyrophosphate and isopentyl pyrophosphate to various carotenoid precursors and carotenoids, especially .beta.-carotene, lycopene, zeaxanthin and cantaxanthin. Processes are also provided for preparing zeaxanthin by culturing a transformed host cell containing an expression cassette that includes a polynucleotide having a DNA sequence which encodes the GGPP synthase of Flavobacterium sp. R1534 (crtE), the prephytoene synthase of Flavobacterium sp. R1534 (crtB), the phytoene desaturase of Flavobacterium sp. R1534 (crtI), the lycopene cyclase of Flavobacterium sp. R1534 (crtY), or the .beta.-carotene hydroxylase of Flavobacterium sp. R1534 (crtZ). The polynucleotide is substantially free of other polynucleotides of Flavobacterium sp. R1534. The process further includes isolating the zeaxanthin from such cells or the culture medium.

Abstract: A process for cultivating Haematococcus for the large scale production of astaxanthin-enriched Haematococcus cells comprises: (a) cultivating said Haematococcus cells under conditions suitable for optimal vegetative growth of said cells, wherein said conditions comprise growing the cells under a light intensity in the range of about 30-140 .mu.mol photons.m.sup.-2.S.sup.-1 and at a temperature of between about 15-28.degree. C.; and (b) collecting the cells grown according to (a) and cultivating them further under conditions suitable for optimal induction and accumulation of astaxanthin in said cells, wherein said conditions comprise inoculating said cells of (a) into a growth solution containing essentially a carbon source and growing said cells at a temperature of below 35.degree. C.

Abstract: Phaffia rhodozyma strains are described which produce greater than 3,000 ppm astaxanthin based on dry yeast solids when cultivated in a volume of nutrient medium of at least about 1,500 liters and containing in excess of 4 percent, preferably in excess of 6 percent, dry yeast solids. These and other strains are cultivated by an improved fermentation method comprising extending the maturation phase of the fermentation by one or more various techniques including exposing the yeast cells to a low-intensity light, slow feeding the cells with a rapidly metabolized energy source, e.g. glucose, and replacing the rapidly metabolized energy source with a slowly metabolized energy source, e.g. gylcerol. The cells of these strains are incorporated into animal feeds, particularly feeds for salmonid fishes, to impart or enhance the red pigmentation of these animals and products made from these animals.

Abstract: An isolated pure culture of a strain of the yeast Phaffia rhodozyma which produces large amounts of astaxanthin, in excess of about 600 micrograms per gram of Phaffia rhodozyma dry matter, as determined by HPLC analysis, is disclosed.

Abstract: A novel Paracoccus species type strain DSM 11574 which produces and secretes carotenoids such as .beta.-carotene, echinenone, .beta.-cryptoxanthin, canthaxanthin, adonirubin, cis-adonixanthin, adonixanthin, astaxanthin and zeaxanthin in vesicles; carotenoid containing vesicles; and a process for production of carotenoids comprising (a) culturing a bacterial species in an nutrient medium including sources of carbon, nitrogen and inorganic substances; and (b) recovering an individual carotenoid pigment or a mixture of carotenoid pigments from the cells, vesicles and/or medium.

Abstract: Phaffia rhodozyma strains are described which produce greater than 3,000 ppm astaxanthin based on dry yeast solids when cultivated in a volume of nutrient medium of at least about 1,500 liters and containing in excess of 4 percent, preferably in excess of 6 percent, dry yeast solids. These and other strains are cultivated by an improved fermentation method comprising extending the maturation phase of the fermentation by one or more various techniques including exposing the yeast cells to a low-intensity light, slow feeding the cells with a rapidly metabolized energy source e.g. glucose, and replacing the rapidly metabolized energy source with a slowly metabolized energy source, e.g. gylcerol. The cells of these strains are incorporated into animal feeds, particularly feeds for salmonid fishes, to impart or enhance the red pigmentation of these animals and products made from these animals.

Abstract: A process for dewatering an aqueous suspension of microalgae is disclosed in which the aqueous suspension of the algae is introduced into a bubble column or a modified bubble column for generating a froth of bubbles and adsorbed algal cells that can be separated from the aqueous suspension. In one advantageous embodiment, the bubble column is a multi-stage loop-flow flotation column that has three loop-flow zones, each of which is defined by a draft tube concentrically mounted in the column to divide each loop-flow zone into a riser and the downcomer. Fine bubbles of gas and brine are in cocurrent upward flow in the riser and in cocurrent downward flow the downcomer. A higher gas holdup is promoted in the riser than in the downcomer, thereby circulating the brine in loop-flow upwardly through the riser and downwardly through the downcomer. Liquid communication between adjacent loop-flow zones is substantially eliminated. A froth enriched in algae is generated that can be separated from the aqueous suspension.

Abstract: DNA sequences are described that encode genes for synthesizing ketocarotenoids such as astaxanthin. The DNA sequences, microorganisms containing them and encoded polypeptides are described. Also described are methods to obtain related sequences and to make host cells that contain such sequences. These genes and methods are useful to impart red coloration during culture of fish and crustaceans, as coloring in food, and as antioxidants.

Abstract: A control method for operating aqueous Haematococcus spp., such as H. pluvialis, microorganism growth processes is disclosed which can maintain viable growth conditions of this microorganism which have heretofore not been easily reproduced in commercially valuable quantities. The primary control parameters are the degree of turbulence in the aqueous growth medium and the scale of the apparatus relative to the scale of the turbulent eddies in vessels which are partially filled with the aqueous medium directly affect conditions which are required for optimum growth: light exposure, nutrient supply, sedimentation rate, bulk temperature, gas exchange rate and cell integrity. These control elements can be cast in terms of the Reynolds number (N.sub.re), pH, temperature, amount of impinging light, and NO.sub.2 concentration, depending upon the operative chlorophyll growth stages of Haematococcus spp. and its photoadaptive stages of producing astaxanthin.

Abstract: The present invention provides Phaffia rhodozyma strains having increased levels of astaxantin and a low percentage of 3-hydroxy-3',4'-didehydro-.beta.,.psi.-caroten-4-one (HDCO). Such strains are obtained by a combination of mutagenesis and selection. The invention further provides a method for obtaining such strains. The invention also provides astaxanthin obtained from a mutagenized strain of Phaffia rhodozyma characterized in that it contains a decreased relative amount of HDCO.

Abstract: Bacterium belonging to a new genus, and a process for production of carotenoid pigment selected from the group consisting of astaxanthin, adonixanthin, .beta.-carotene, echinenone, canthaxanthin and zeaxanthin, comprising the steps of culturing a bacterium capable of producing at least one of the carotenoid pigments, and recovering an individual carotenoid pigments.The producer bacterium belongs to new genus.According to the present process various carotenoids can be produced in an industrial scale. By the present process (3S, 3'S)-astaxanthin can be produced in an almost 100% purity.

Abstract: A process for isolating and purifying lycopene crystals from a biological lycopene source is disclosed. A lycopene-containing oleoresin is saponified in a composition of propylene glycol and aqueous alkali to form lycopene crystals. Crystallization is achieved without the use of added organic solvents. The crystals are isolated and purified. The substantially pure lycopene crystals so obtained are suitable for human consumption and can be used as a nutritional supplement and as an additive in food.

Abstract: A method is disclosed for making the 3R-3'R stereoisomer of zeaxanthin as a sole detectable or heavily dominant (more than about 90%) stereoisomer, for human ingestion. Zeaxanthin, a yellowish pigment which is naturally present in macular cells in the center of the human retina, absorbs blue and near-ultraviolet light radiation, thereby protecting the retinal cells against phototoxic damage. Zeaxanthin preparations which contain only the desired R--R isomer can be produced by a strain of Flavobacterium multivorum cells (ATCC accession number 55238). These cells (and other cells transformed with their zeaxanthin-producing genes) do not create any detectable quantities of the undesired S--S or S-R isomers of zeaxanthin, and they do not synthesize significant quantities of other carotenoids which might compete against zeaxanthin for alimentary uptake after oral ingestion.

Abstract: The therapeutic or dietetic use of a preparation based on phospholipids extracted from pig brains for treating aging disorders consisting of memory disorders, attention disorders, intellectual efficacy disorders, humor and affectivity disorders, and thinking difficulty in a human is disclosed. The purified brain phospholipids are used alone or in combination with carriers and diluents enabling oral delivery.

Abstract: Disclosed are the following DNA strands relating to the synthesis of keto group-containing xanthophylls such as astaxanthin and the like, and the techniques relating to the production of xanthophylls by genetic engineering:A DNA strand having a nucleotide sequence which encodes a polypeptide having an enzyme activity for converting a methylene group at the 4-position of .beta.-ionone ring into a keto group.A DNA strand having a nucleotide sequence which encodes a polypeptide having an enzyme activity for converting a methylene group at the 4-position of a 3-hydroxy-.beta.-ionone ring into a keto group.A DNA strand having a nucleotide sequence which encodes a polypeptide having an enzyme activity for adding a hydroxyl group to the 3-carbon of a 4-keto-.beta.-ionone ring.It is possible to produce a variety of xanthophylls such as canthaxanthin, astaxanthin and the like by introducing the DNA strands into an appropriate microorganism such as Escherichia coli and the like.

Abstract: Methods are disclosed for dewatering an aqueous suspension of microalgae, especially Dunaliella salina, with a Jameson cell. The algal cells are ruptured sufficiently to promote froth flotation of the cells from the suspension. The algal suspension is introduced into the top of the Jameson cell downcomer and passed through an orifice plate therein to generate a free liquid jet of the suspension. The jet is plunged downwardly through a gas and the gas is transported with the jet through a gas and liquid interface into the algal suspension. The momentum of the jet is dissipated, which disperses the gas into fine bubbles. The fine bubbles are intimately contacted with the aqueous suspension in cocurrent downward flow in a collection zone to absorb the algal bodies onto the surfaces of the bubbles. A froth of bubble and algal agglomerates is formed and collapsed to obtain a dewatered algal concentrate.

Abstract: The present invention relates to astaxanthin-containing zooplankton such as Brachionus pricatilis obtained by culturing, a method for culturing the zooplankton in a liquid containing astaxanthin and a method for breeding fry fish by feeding the above astaxanthin-containing zooplankton.The survival rate in breeding fry fish may be remarkably improved by feeding the astaxanthin-containing zooplankton to fry fish.

Abstract: Thermus aquaticus biovar. Nov. SK542 (FERM BP-3382) is an absolute aerobic bacteria. It grows at temperature limit of 40.degree.-82.degree. C. in a normal concentration medium, but its best growth is achieved at 72.degree.-76.degree. C. It produces protein decomposing enzymes functional at a temperature of 75.degree.-85.degree. C. and active in a wide pH range of 4.0-11.3, and a yellow pigment of carotenoid groups. A method for improving the quality of soil comprising applying to the soil a biologically pure culture of an absolute aerobic bacterium Thermus aquaticus biovar. nov. SK542 (FERM BP-3382) having a growth temperature limit of 40.degree.-82.degree. C. in a normal concentration medium, a growth optimum temperature of 72.degree.-76.degree. C., and producing protein decomposing enzymes functional at temperature range of 75.degree.-85.degree. C. and being active in a wide pH range of 4.0-11.3, and a yellow pigment of carotenoid.

Abstract: An isolated pure culture of a strain of Phaffia rhodozyma which produces astaxanthin in an amount of at least 600 .mu.g per g Phaffia rhodozyma dry matter, as determined by HPLC analysis.

Abstract: An isolated pure culture of a strain of Phaffia rhodozyma which produces astaxanthin in an amount of at least 600 .mu.g per g Phaffia rhodozyma dry matter, as determined by HPLC analysis.

Abstract: The present invention relates to a novel enzymatic process for the preparation of flavors, in particular of the ionones and C.sub.6 to C.sub.10 aldehydes. The invention discloses a process for the preparation of optically active alpha ionone, beta ionone, C.sub.6 aldehydes such as n-hexanal, trans-2-hexanal, C.sub.10 aldehydes such as trans-2, cis-4-decadidienal, and trans-2, trans-4-decadienal. The invention also relates to a process for the production of alcohols, particularly C.sub.6 and C.sub.10 alcohols.

Abstract: The subject of the present invention is Phaffia rhodozyma mutants blocked in the step of conversion of .beta.-carotene in the pathway of biosynthesis of astaxanthin.The present invention also relates to a process for producing .beta.-carotene using these mutants, for purposes of use in the preparation of foodstuffs or cosmetic or pharmaceutical products.

Abstract: DNA segments encoding the Erwinia herbicola enzymes geranylgeranyl pyrophosphate (GGPP) synthase, phytoene synthase, phytoene dehydrogenase-4H, lycopene cyclase, beta-carotene hydroxylase, and zeaxanthin glycosylase, DNA variants and analogs thereof encoding an enzyme exhibiting substantially the same biological activity, vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes, zeaxanthin and zeaxanthin diglucoside by recombinant DNA technology in transformed host organisms are disclosed.

Abstract: An isolated pure culture of a strain of Phaffia rhodozyma which produces astaxanthin in an mount of at least 600 .mu.g per g Phaffia rhodozyma dry matter, as determined by HPLC analysis.

Abstract: DNA segments encoding the Erwinia herbicola enzymes geranylgeranyl pyrophosphate (GGPP) synthase, phytoene synthase, phytoene dehydrogenase-4H and lycopene cyclase, vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes and beta-carotene by recombinant DNA technology in transformed host organisms are disclosed.

Abstract: The present invention provides Phaffia rhodozyma strains having increased levels of astaxantin and a low percentage of 3-hydroxy-3',4'-didehydro-.beta.,.psi.-caroten-4-one (HDCO). Such strains are obtained by a combination of mutagenesis and selection. The invention further provides a method for obtaining such strains. The invention also provides astaxanthin obtained from a mutagenized strain of Phaffia rhodozyma characterized in that it contains a decreased relative amount of HDCO.

Abstract: The present invention discloses intersexual heterozygous Phycomyces strains. These strains show an improved .beta.-carotene productivity and an increased stability when compared with both the wild type and heterokaryotic Phycomyces strains. The present invention also discloses a method for obtaining Phycomyces transformants without using selection markers. Furthermore, a process is provided for manufacturing .beta.-carotene comprising the use of intersexual heterozygous Phycomyces strains in a submerged culture.

Abstract: Bacterium belonging to a new genus, and a process for production of carotenoid pigment selected from the group consisting of astaxanthin, adonixanthin, .beta.-carotene, echinenone, canthaxanthin and zeaxanthin, comprising the steps of culturing a bacterium capable of producing at least one of the carotenoid pigments, and recovering an individual carotenoid pigments.The producer bacterium belongs to new genus.According to the present process various carotenoids can be produced in an industrial scale. By the present process (3S, 3'S)-astaxanthin can be produced in an almost 100% purity.

Abstract: An isolated pure culture of a strain of Phaffia rhodozyma which produces astaxanthin in an amount of at least 600 .mu.g per g Phaffia rhodozyma dry matter, as determined by HPLC analysis.

Abstract: A process for production of canthaxanthin comprising the steps of culturing a microorganism capable of producing canthaxanthin and belonging to the genus Corynebacterium, such as Corynebacterium sp. SQH 348 (FERM BP-4284), and recovering canthaxanthin from the culture.

Abstract: A method of growing microalgae is described, which uses the outdoor sunlight as a source of energy. Growth is confined to an assembly of vertical, transparent tubes through which nutrient and air is carried with carbon dioxide. The microalgae is periodically harvested from the tubes.

Abstract: A process for production of canthaxanthin comprising the steps of culturing a microorganism capable of producing canthaxanthin and belonging to the genus Corynebacterium, such as Corynebacterium sp. SQH 348 (FERM BP-4284), and recovering canthaxanthin from the culture.

Abstract: Phaffia rhodozyma strains are described which produce greater than 3,000 ppm astaxanthin based on dry yeast solids when cultivated in a volume of nutrient medium of at least about 1,500 liters and containing in excess of 4 percent, preferably in excess of 6 percent, dry yeast solids. These and other strains are cultivated by an improved fermentation method comprising extending the maturation phase of the fermentation by one or more various techniques including exposing the yeast cells to a low-intensity light, slow feeding the cells with a rapidly metabolized energy source, e.g. glucose, and replacing the rapidly metabolized energy source with a slowly metabolized energy source, e.g. gylcerol. The cells of these strains are incorporated into animal feeds, particularly feeds for salmonid fishes, to impart or enhance the red pigmentation of these animals and products made from these animals.

Abstract: Disclosed are DNA sequences which are useful for the synthesis of carotenoids such as lycopene, .beta.-carotene, zeaxanthin or zeaxanthin-diglucoside, that is, DNA sequences encoding carotenoid biosynthesis enzymes. These DNA sequences are the sequences 1-6 shown in the specification.Also disclosed is a process for producing a carotenoid or a carotenoid related compound which is selected from the group consisting of geranylgeranyl pyrophosphate, phytoene, lycopene, .beta.-carotene, zeaxanthin and zeaxanthin-diglucoside, which comprises transforming a host with at least one of the DNA sequences 1-6 described above and culturing the transformant.

Abstract: Zeaxanthin is produced using Flavobacterium multivorum. The process and the nutrient medium used in the process provide greater zeaxanthin and cell yields per liter, at a lower cost, and more rapidly than known methods and microorganisms. Biomass compositions containing the microorganism of this invention are also disclosed.

Abstract: The present invention is directed toward a method for producing beta-carotene using a mated culture of Mucorales fungi. The method includes mutating and selecting negative (minus mating type) and positive (plus mating type) Mucorales fungal microorganisms, culturing the selected negative and positive microorganisms in an effective medium to form a mated culture that produces beta-carotene, and recovering beta-carotene therefrom. The present invention provides mated cultures that overproduce beta-carotene and is also directed to certain negative and positive microorganisms used to overproduce beta-carotene. The present invention also provides beta-carotene formulations produced by the claimed method, and the use of such formulations, for example, to enhance pigmentation, to reduce damage caused by reactive oxygen species or phototoxic molecules, to prevent or treat cancer or cardiovascular disease, to provide a Vitamin A supplement, to enhance lactation, and to increase fertility.

Abstract: The present invention is directed toward a process for producing zeaxanthin which includes mutating algal microorganisms, selecting from said mutated microorganisms a microorganism capable of producing zeaxanthin, culturing the selected microorganism in an effective medium to produce zeaxanthin, and recovering zeaxanthin produced by the selected microorganism. The present invention also provides microorganisms capable of producing zeaxanthin, formulations containing zeaxanthin produced by the disclosed process and the use of such formulations to enhance pigmentation and to reduce damage caused by reactive oxygen species and phototoxic molecules.

Abstract: An isolated pure culture of a strain of Phaffia rhodozyma which produces astaxanthin in an amount of at least 600 .mu.g per g Phaffla rhodozyma dry matter, as determined by HPLC analysis.

Abstract: An economical process for in vivo production of the pigment astaxanthin, and particularly a process for enhancing astaxanthin content of cultures of microorganisms of genus Phaffia, the process comprising culturing a microorganism of genus Phaffia in a nutrient medium containing an antibiotic, a cytochrome B inhibitor, or a terpenoid synthetic pathway inhibitor, cultivating surviving pigment enhanced microorganisms, and harvesting the yeast.

Abstract: The present invention is directed toward a method for producing beta-carotene using negative (minus mating type) Mucorales fungal microorganisms. The method includes mutating and selecting negative Mucorales fungal microorganisms, culturing the selected negative microorganisms in an effective medium to produce beta-carotene, and recovering beta-carotene therefrom. The present invention provides negative microorganisms that overproduce beta-carotene, beta-carotene formulations produced by the disclosed method, and the use of such formulations to enhance pigmentation, to reduce damage caused by reactive oxygen species or phototoxic molecules, to prevent or treat cancer or cardiovascular disease, to provide a Vitamin A supplement, to enhance lactation, and to increase fertility.

Abstract: Zeaxanthin is produced using Flavobacterium multivorum. The process and the nutrient medium used in the process provide greater zeaxanthin and cell yields per liter, at a lower cost, and more rapidly than known methods and microorganisms. Biomass compositions containing the microorganism of this invention are also disclosed.

Abstract: The production and use of a high-intensity red natural colorant prepared from an anthocyanin derived from the cell line of Daucus carota (carrot) is described. The production of the high-intensity red natural colorant comprises the cultivation of Daucus carota in a cell tissue culture. Carrot cells are extracted from the cell tissue culture after subculturing and a suitable growth period. The extract is then purified in order to isolate the anthocyanin found in the carrot cells. The anthocyanin is then concentrated to produce the red natural colorant of the invention. The natural colorant produced is stable over a wide pH range under various conditions. The natural colorant is useful as a coloring agent for food products, cosmetics, and pharmaceuticals.

Abstract: The production and use of a high-intensity red natural colorant prepared from an anthocyanin derived from the cell line of Daucus carota (carrot) is described. The production of the high-intensity red natural colorant comprises the cultivation of Daucus carota in a cell tissue culture. Carrot cells are extracted from the cell tissue culture after subculturing and a suitable growth period. The extract is then purified in order to isolate the anthocyanin found in the carrot cells. The anthocyanin is then concentrated to produce the red natural colorant of the invention. The natural colorant produced is stable over a wide pH range under various conditions. The natural colorant is useful as a coloring agent for food products, cosmetics, and pharmaceuticals.

Abstract: A method of extracting carotenoid, tetrapyrrole and porphyrin from vegetable sources of such pigments, especially algae, as described. The method entails production of water soluble and oil soluble extracts and includes an extractive procedure for rendering carotenoids water soluble. The pigment extracts can be used as nutritional or medicinal compositions.

Abstract: A novel complex designated herein as rigolettone complex is produced by fermentation of Streptomyces aburaviensis strain C-38,242 (ATCC 39290). The complex and its purified bioactive components, jildamycin and mantuamycin, exhibit antitumor activity in mouse tumor systems.

Abstract: Process for extracting dyestuffs and particularly a blue pigment, called phycocyanine, from cyanophyceae algae, particularly of the Spirulina species, comprising contacting said algae with a first aqueous phase containing calcium ions, separating therefrom the mass of algae and contacting it with a second aqueous phase of alkaline character, separating the algae therefrom and extracting phycocyanine from said second aqueous phase by subjecting the same to an ultrafiltration step for concentrating the solution and then to a drying step.

Abstract: This invention refers to the obtention of strains of the fungus Phycomyces blakesleeanus accumulated by the .beta.-carotene pigments in amounts which permit the use of these strains in the production on an industrial scale of said pigment. By genetic manipulations, strains simultaneously and constitutively incorporating various stimulating effects have been isolated. The resulting strains produce .beta.-carotene in a proportion of approximately 2.5% of its dry weight, which constitutes almost 1000 times more than that which the natural microorganism would produce under the same conditions. This production level, together with the simplicity in cultivating and manipulating same, makes them interesting in the development of an industrial process for producing .beta.-carotene by fermentation. The .beta.-carotene produced by fermentation, besides its low cost, would have the advantage of not being contaminated with the reagents and catalysts used in the organic synthesis.