Abstract: The invention relates to coryneform bacteria which have, in addition to at least one copy, present at the natural site (locus), of an open reading frame (ORF), gene or allele which codes for the synthesis of a protein or an RNA, in each case a second, optionally third or fourth copy of this open reading frame (ORF), gene or allele at in each case a second, optionally third or fourth site in a form integrated into the chromosome and processes for the preparation of chemical compounds by fermentation of these bacteria.

Abstract: The present invention relates to the DNA sequence for eukaryotic genes encoding &egr; cyclase isolated from romaine lettuce as well as vectors containing the same and hosts transformed with said vectors. The present invention provides methods for controlling the ratio of various carotenoids in a host and to the production of novel carotenoid pigments. The present invention also provides a method for treating disease by administering carotenoids obtained from transformed hosts, or by administering a composition containing the transformed hosts.

Abstract: The invention relates to a process for the recovery of minor components and refining of vegetable oils and fats from crude vegetable oils and fats. The said invention describes the following process: A process for the recovery of minor components and refining of vegetable oils and fats from crude vegetable oils and fats without destroying naturally occurring components, said process comprising the steps of: a) removal of polar components from the crude vegetable oils and fats using lower alkyl alcohol or any lower alkyl alcohol-water mixture; b) removal of alcohol from the product obtained in step (a) by distillation; c) addition of suitable quantity of bleaching earth to the product obtained in step (b) at normal bleaching temperature followed by filtration; and d) deodorization of the product obtained in step (c) at a low temperature.

Abstract: A lycopene &egr; cyclase from spinach is described, as is production of lutein in microorganisms by expression of the lycopene &egr; cyclase.

Abstract: Nucleic acid sequences encoding ∈-cyclase, isopentenyl pyrophosphate isomerase and &bgr;-carotene hydroxylase as well as vectors containing the same and hosts transformed with the vectors. Methods for controlling the ratio of various carotenoids in a host and for the production of novel carotenoid pigments. The present invention also provides a method for screening for eukaryotic genes encoding carotenoid biosynthesis, and for modifying the disclosed enzymes.

Abstract: A process is described for preparing dry powders of one or more oxygenated carotenoids by a) dispersing one or more oxygenated carotenoids in an aqueous molecular dispersion or colloidal dispersion of a protecting colloid and b) converting the dispersion formed into a dry powder by removing the water and any solvents additionally used and drying, in the presence or absence of a coating material, which comprises using as protecting colloid in process step a) at least one partially hydrolyzed soybean protein having a degree of hydrolysis greater than 5%.

Abstract: The present invention relates to an isolated DNA sequence encoding an enzyme in the mevalonate pathway or the pathway from isopentenyl pyrophosphate to farnesyl pyrophosphate. Vectors and plasmids including such DNA are also set forth. The invention also includes host cells transformed by such DNAs, or vectors or plasmids containing such DNAs. A process for the production of isoprenoids and carotenoids using such transformed host cells is also provided.

Abstract: Genes isolated from Methylomonas sp. 16a have been determined to play a role in the carotenoid biosynthetic pathway. Specifically, crtN2 gene has the ability to produce omega-aldehyde functional groups on carotenogenic substrates, while the ald gene produced omega carboxyl functional groups. These genes will be useful for production of high levels of functionalized carotenoid compounds, especially those produced in microorganisms which metabolize single carbon substrates.

Abstract: A method for producing beta-carotene and carotene-related pigments in which a plurality of thermophilic microorganisms is collected and screened for the production of pigments. Those pigment-producing thermophilic microorganisms having yellow, red or orange coloration are identified and separated from the collection of thermophilic microorganisms. Thereafter, the selected pigment-producing thermophilic microorganisms are mutated by non-recombinant means to enhance pigment production, forming a mutant pigment-producing thermophilic microorganism. In accordance with one embodiment of this invention, a gene of interest suitable for producing a protein of interest is introduced into the mutant pigment-producing thermophilic microorganism, resulting in over-production of the carotene pigment and the protein of interest. Also disclosed are suitable plasmids and expression vectors suitable for use in the method of this invention.

Abstract: Novel proteins of Flavobacterium 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 &bgr;-carotene, lycopene, zeaxanthin and cantaxanthin.

Abstract: Isolated nucleic acid molecules, designated TCMRP nucleic acid molecules, which encode novel TCMRPs from e.g. Phycomitrella patens are described. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing TCMRP nucleic acid molecules, and host cells into which the expression vectors have been introduced. The invention still further provides isolated TCMRPs, mutated TCMRPs, fusion proteins, antigenic peptides and methods for the improvement of production of a desired compound from transformed cells, organisms or plants based on genetic engineering of TCMRP genes in these organisms.

Abstract: Genes have been isolated from Pantoea stewartii encoding geranylgeranyl pyrophosphate (GGPP) synthase (crtE), phytoene synthase (crtB), phytoene desaturase(crtI), lycopene cyclase(crtY), &bgr;-carotene hydroxylase(crtZ), and zeaxanthin glucosyl transferase (crtX) activity. The genes and their products are useful for the conversion of phytoene to the carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes and &bgr;-carotene by recombinant DNA technology in transformed host organisms are disclosed.

Abstract: Isolated polynucleotides encoding polypeptides having the activity of enzymes in the mevalonate pathway are provided. These sequences are useful for recombinantly producing isoprenoid compounds, such as carotenoids, in particular zeaxanthin. Expression vectors, cultured cells, and methods of making isoprenoid compounds are also provided.

Abstract: A process for the isolation of desired compound(s) from a microbial biomass is disclosed, wherein the microbial biomass (which, if necessary, is pretreated to give a dry matter content of from 25 to 80%) is granulated (e.g. by extrusion) and then dried to a dry matter content of at least 80%. The granulation of the biomass to granules significantly eases subsequent drying of the biomass (which can be stored as dried granules) and gives higher yields on extraction of the compound(s).

Abstract: Genes have been isolated from Rhodococcus and Deinococcus which encode a specific lycopene &bgr;-cyclase capable of converting acyclic carotenoids with at least one &psgr;-end group to the corresponding asymmetric carotenoid containing a single &bgr;-ionone ring end group. The genes are new. Transformed host cells expressing the present genes and methods for the bio-conversion of acylic carotenoid substrates to corresponding asymmetric carotenoid are also provided.

Abstract: The present invention relates to a crystalline carotenoid compound, such as &bgr;-carotene, with a purity of at least 95% and with substantially no solvent enclosed in the crystal lattice. The present invention further describes a process to prepare such a highly pure crystalline carotenoid compound from microbial biomass, without the use of a solvent extraction and/or an anti-solvent crystallization process.

Abstract: The present invention relates to an isolated DNA sequence encoding an enzyme in the mevalonate pathway or the pathway from isopentenyl pyrophosphate to farnesyl pyrophosphate. Vectors and plasmids including such DNA are also set forth. The invention also includes host cells transformed by such DNAs, or vectors or plasmids containing such DNAs. A process for the production of isoprenoids and carotenoids using such transformed host cells is also provided.

Abstract: A method of chromatographic isolation for non-glyceride components (squalenes, carotenes, vitamin E, sterols and/or the like) from a non-glyceride components-comprising compound by the steps of a. introducing the non-glyceride components-comprising compound onto a selective adsorbent to allow an adsorption of the non-glyceride components, and subsequently b. desorbing the non-glyceride components from the adsorbent, wherein the adsorption and/or desorption of the non-glyceride components is carried out under a supercritical fluid environment.

Abstract: Preparations are disclosed containing the 3R-3′R stereoisomer of zeaxanthin, packaged for oral ingestion by humans as a therapeutic drug or nutritional supplement. Zeaxanthin is a yellow carotenoid pigment found in the macula (in the center of the human retina), which helps protect retinal cells against phototoxic damage. The R-R stereoisomer can be prepared by fermenting cells, such as Flavobacterium multivorum (ATCC 55238), which do not create any detectable quantity of the undesired and potentially toxic S-S or S-R isomers, and which do not synthesize any other carotenoids. The R-R isomer can be concentrated, in large quantities and at low cost, into a viscous oily fluid containing about 5 to 20% zeaxanthin, by means of a simple solvent extraction process. This oily fluid can be mixed with a carrier such as vegetable oil and enclosed within a digestible capsule, comparable to a conventional capsule containing Vitamin E.

Abstract: A ketolase gene has been isolated from Rhodococcus erythropolis AN12 strain encoding a carotenoid modification enzyme of the carotenoid biosynthetic pathway. The gene and gene product are the first isolated from a Rhodococcus strain. Six conserved amino acid motifs have been identified as the characteristic of this type of ketolase enzymes. The gene and gene product of the present invention may be used in a variety of ways for the production of keto-carotenoid compounds in a variety of organisms.

Abstract: Isolated gene cluster involved in canthaxanthin biosynthesis, which comprises a polynucleotide wherein:

Abstract: Nucleic acid sequences coding for a protein having &bgr;,&bgr;-carotene 15,15′-monooxygenase activity and their uses in diagnostics, the synthesis of vitamin A and methods for the introduction of the &bgr;,&bgr;-carotene 15,15′-monooxygenase cDNA into host cells are disclosed.

Abstract: The present invention relates to a process for extracting astaxanthin pigment from the yeast cells of Phaffia rhodozyma comprising the steps of i) cultivating the yeast, ii) treating yeast culture suspension with microwave to destroy the cell walls and microbodies, iii) drying it or extracting astaxanthin pigment using the solvent selected from the group consisting of ethanol, methanol, acetone and mixture of them. Further, the present invention also relates to the astaxanthin extracted by above method.

Abstract: The present invention is directed to genetic materials useful for the preparation of astaxanthin from beta-carotene, such as polypeptides having astaxanthin synthase activity, DNA fragments coding for astaxanthin synthase, recombinant organisms and the like. Those novel genetic materials may be derived from Phaffia rhodozyma. The present invention also provides a process for the production of astaxanthin.

Abstract: A purified nucleic acid sequence which encodes for a protein having ketolase enzyme activity and has the nucleic acid sequence of SEQ ID NO: 1 or 3, or has a sequence which encodes the amino acid sequence of SEQ ID NO: 2 or 4, as well as vectors and host cells containing them. Methods of use of the nucleic acid sequences to produce ketocarotenoid in host cells and methods of use of the nucleic acid sequences to modify the production of carotenoids in a host cell are included.

Abstract: A method for production of high amounts of carotenoids in plants, bacteria or cells. Carotenoid-associated proteins CHRC and CHRD isolated from corollas of Cucumis sativus, chrc gene, CHRC deduced sequence, the molecular cloning, cDNA and RNA. A new CHRC promoter able to direct expression for foreign genes. The cloned promoter comprising approximately 3.5 Kb of the CHRC upstream region.

Abstract: The invention features retina-derived (retinal endothelial or retinal epithelial pigment) cell lines with extended life-span and capable of being implanted in the retina and of carrying a therapeutic substance to the eye and to the central nervous system. Such lines can also be used as a model for studying blood central nervous system interfaces. These lines are derived from primary retinal cultures selected from the group consisting of primary retinal endothelial cells and primary retinal epithelial cells, comprise a polynucleotide containing an oncogene, which polynucleotide is optionally associated with at least one selection gene, and have the morphological characteristics and at least the expression characteristics of the surface antigens of corresponding primary cultures.

Abstract: A process for producing carotenoids, which involves cultivating a microorganism obtained by treating a parent microorganism that produces carotenoids under conditions that induce a reduction in alternative oxidase activity and selecting a microorganism with enhanced carotenoid productivity, a method for obtaining the microorganism with enhanced carotenoid productivity, and the microorganism itself.

Abstract: The present invention provides a method for changing production ratios of carotenoid compounds in a process of microbiological production of a plurality of carotenoid compounds. By controlling the concentration of dissolved oxygen in the culture during cultivation, production ratios of carotenoid compounds such as astaxanthin, adonixanthin, &bgr;-carotene, echinenone, canthaxanthin, zeaxanthin, &bgr;-cryptoxanthin, 3-hydroxyechinenone, asteroidenone and adonirubin, are changed.

Abstract: The present invention describes the DNA sequence for eukaryotic genes encoding &egr; cyclase, isopentenyl pyrophosphate (IPP) isomerase and &bgr;-carotene hydroxylase as well as vectors containing the same and host cells transformed with said vectors. The &egr; cyclase and &bgr;-carotene hydroxylase genes disclosed include those from A. thaliana; the IPP isomerase genes disclosed include those from A. thaliana, H. pluvialis, and marigold. The present invention also provides methods for controlling the ratio of various carotenoids in a host cell and for the production of novel carotenoid pigments. The present invention also provides a method for screening for eukaryotic genes encoding carotenoid biosynthesis enzymes.

Abstract: Novel proteins of microorganism E-396 (FERM BP-4283) and the DNA sequences which encode these proteins have been discovered to provide an improved biosynthetic pathway from farnesyl pyrophosphate and isopentyl pyrophosphate to various carotenoids, especially zeaxanthin, astaxanthin, adonixanthin and canthaxanthin.

Abstract: A method for the production of carotenoid compounds is disclosed. The method relies on the use of microorganisms which metabolize single carbon substrates for the production of carotenoid compounds in high yields.

Abstract: A method of increasing a fraction of free carotenoids in a source of carotenoids in which at least some of the carotenoids are fatty acid esterified carotenoids is disclosed. The method is effected by contacting the source of carotenoids with an effective amount of an esterase under conditions effective in deesterifying the fatty acid esterified carotenoids, thereby increasing the fraction of free carotenoids in the source of carotenoids.

Abstract: A process is provided for producing a carotenoid, which process includes cultivating a recombinant organism having a gene for one or more active oxygen species-quenching factor(s) that is substantially disrupted with a disruption cassette specific to the gene, and recovering carotenoids from the culture.

Abstract: Novel proteins of Flavobacterium sp. R1534 and the DNA sequences which encode these proteins are disclosed which provide an improved biosynthetic pathway from famesyl pyrophosphate and isopentyl pyrophosphate to various carotenoid precursors and carotenoids, especially &bgr;-carotene, lycopene, zeaxanthin and cantaxanthin.

Abstract: Nucleic acid sequences encoding ∈-cyclase, isopentenyl pyrophosphate isomerase and &bgr;-carotene hydroxylase as well as vectors containing the same and hosts transformed with the vectors. Methods for controlling the ratio of various carotenoids in a host and for the production of novel carotenoid pigments. The present invention also provides a method for screening for eukaryotic genes encoding carotenoid biosynthesis, and for modifying the disclosed enzymes.

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 the isolation of crystalline carotenoid compound from microbial biomass comprising disrupting the microbial cell walls, separating cellular debris from the carotenoid-containing residue, washing one member of the group consisting of the microbial biomass, the disrupted cell mass and the carotenoid-containing reside with a solvent to remove lipid, suspending the obtained carotenoid crystals in water to float the crystals, recovering the crystals and, optionally, further purifying the crystals which avoids the use of large amounts of solvent of carotenoid extraction process.

Abstract: The present invention relates, in general, to a biotechnological method for production of (3S,3′S) astaxanthin. In particular, the present invention relates to a peptide having a &bgr;-C-4-oxygenase activity; a DNA segment coding for this peptide; an RNA segments coding for this peptide; a recombinant DNA molecule comprising a vector and the DNA segment; a host cell or organism containing the above described recombinant DNA molecule or DNA segment; and to a method of biotechnologically producing (3S,3′S) astaxanthin or a food additive containing (3S,3′S) astaxanthin, using the host.

Abstract: The present invention relates to engineering new biosynthetic pathways into microorganisms, in particular biosynthetic carotenoid pathways. New and improved catalytic functions of metabolic pathways are created by, for example, site-specific mutation or gene shuffling techniques, to provide for efficient biosynthesis of carotenoids. By applying the described directed evolution techniques, almost any carotenoid could be produced, in a host cell, from one or a few sets of genes. In addition, the described techniques are useful for creating gene or protein libraries for new and uncharacterized carotenoids.

Abstract: The present invention describes a process to recover highly pure 2-carotene crystals from a crude crystal preparation obtained from a natural source. To remove impurities, the crude crystals are stirred in a solvent in which 2-carotene has a low solubility, whereupon the crystals are filtered off and washed with fresh solvent. Using the process of the invention, a natural crystalline 2-carotene preparation with a very high purity is obtained, a purity comparable to that of synthetic 2-carotene.

Abstract: Disclosed is a method for producing an effective amount of one or more carotenoid, xanthophyll, or apo-carotenoid pigments from a microorganism from the Order Thraustochytriales. Suitable microorganisms put forth are of the genus Thraustochytrium and Schizochytrium. Preferred culture conditions and mediums are discussed. Examples are given of production of specific pigments, such as &bgr;-carotene, lutein, adonirubin, canthaxanthin, and astaxanthin, as are examples of methods of isolation of the pigment from the microorganism. Also disclosed is a food product that contains a microorganism from the Order Thraustochytriales, the microorganism having produced internally an effective amount of one or more carotenoid, xanthophyll, or apo-carotenoid pigments. The food product is used in a method for delivering one or more of the pigments to an animal or human for either pharmaceutical, nutritional or coloration purposes, by feeding the food product to the animal or human.

Abstract: A method of prophylactic treatment of mastitis in mammalian, including human, mothers, is described. The method comprises administration of a prophylactically effective dosage of a human or veterinary medicament containing at least one type of xanthophylls, such as astaxanthin, to said mothers. Preferably, the astaxanthin exists in a form esterified with fatty acids, e.g. in the form of algae meal produced by culturing of the alga Haematococcus sp. Further, use of at least one type of xanthophylls, such as astaxanthin, for the preparation of a human or veterinary medicament for the prophylactic treatment of mastitis is mammalian, including human, mothers is disclosed.

Abstract: The present invention provides a method for preparing astaxanthin from zeaxanthin. Specifically, the present invention provides a method for said conversion using a halogenating agent with the salt of chloric or bromic acid in an inert solvent.

Abstract: A mutant prenyl diphosphate synthase having a modified amino acid sequence of prenyl diphosphate synthase, wherein the amino acid residue located at the eighth position in the N-terminal direction from D of the N-terminal of the Asp-rich domain DDXX(XX)D (in the sequence X denotes any amino acid and the two X's in the parentheses may not be present) present in the region II has been substituted by another amino acid, or both of the amino acid residue located at the fifth position in the N-terminal direction from D of the N-terminal and the amino acid residue located at the eighth position in the N-terminal direction from D of the N-terminal have been substituted by other amino acids.

Abstract: Novel proteins of microorganism E-396 (FERM BP-4283) and the DNA sequences which encode these proteins have been discovered to provide an improved biosynthetic pathway from farnesyl pyrophosphate and isopentyl pyrophosphate to various carotenoids, especially zeaxanthin, astaxanthin, adonixanthin and canthaxanthin.

Abstract: The present invention relates to an isolated DNA sequence encoding an enzyme in the mevalonate pathway or the pathway from isopentenyl pyrophosphate to farnesyl pyrophosphate (i.e., HMG-CoA synthase, HMG-CoA reductase, mevalonate kinase, mevalonate pyrophosphate decarboxylase, and FPP synthase). Vectors and plasmids including such DNA, are also set forth. The invention also includes host cells transformed by such DNAs, or vectors or plasmids containing such DNAs. A process for the production of isoprenoids and carotenoids using such transformed host cells is also provided.

Abstract: A method for inhibiting the activity of the enzyme 5&agr;-reductase in a human subject is provided which comprises administering to the subject a composition comprising the carotenoid astaxanthin. Administration of the composition to inhibit the enzyme is useful to prevent and treat benign prostate hyperplasia (BPH) and prostate cancer in human males.

Abstract: The present invention provides a &bgr;-carotene hydroxylase and a DNA coding for the enzyme. The DNA of the invention codes for the following recombinant protein (a) or (b): (a) a protein consisting of the amino acid sequence as shown in SEQ ID NO: 2; (b) a protein which consists of the amino acid sequence as shown in SEQ ID NO: 2 having deletion, substitution or addition of one or several amino acids and which has &bgr;-carotene hydroxylase activity.

Abstract: Novel proteins of Flavobacterium 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 &bgr;-carotene, lycopene, zeaxanthin and cantaxanthin.