Abstract: A method to increase carotenoid production in carotenogenic microbial host cells is provided by down-regulating or disrupting glycogen synthesis. Disruption of glycogen synthase activity in a carotenogenic microbial host cell significantly increased carotenoid production. Carotenogenic microorganisms are also provided that have been optimized for the production of carotenoid compounds through the down-regulation and/or disruption of glycogen synthase activity.

Abstract: The present invention provides recombinant DNA comprising a transcription promoter and a downstream sequence to be expressed, in operable linkage therewith, wherein the transcription promoter comprises a region found upstream of the open reading frame of a highly expressed Phaffia gene, preferably a glycolytic pathway gene, more preferably the gene coding for Glyceraldehyde-3-Phosphate Dehydrogenase. Further preferred recombinant DNAs according to the invention contain promoters of ribosomal protein encoding genes, more preferably wherein the transcription promoter comprises a region found upstream of the open reading frame encoding a protein as represented by one of the amino acid sequences depicted in any one of SEQIDNOs: 24 to 50.

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 method for the production of aryl-carotenoids is provided through bioconversion of cyclic carotenoids having at least one ?-ionone ring. Expression of a heterologous gene encoding a carotene desaturase (crtU) enzyme in a host cell that produces a suitable cyclic carotenoid substrate effect the production of aryl carotenoids.

Abstract: The present invention provides methods of producing an isoprenoid or an isoprenoid precursor in a genetically modified host cell. The methods generally involve modulating the level of hydroxymethylglutaryl-CoA (HMG-CoA) in the cell, such that the level of HMG-CoA is not toxic to the cell and/or does not substantially inhibit cell growth, but is maintained at a level that provides for high-level production of mevalonate, IPP, and other downstream products of an isoprenoid or isoprenoid pathway, e.g., polyprenyl diphosphates and isoprenoid compounds. The present invention further provides genetically modified host cells that are suitable for use in a subject method. The present invention further provides recombinant nucleic acid constructs for use in generating a subject genetically modified host cell, including recombinant nucleic acid constructs comprising nucleotide sequences encoding one or more mevalonate pathway enzymes, and recombinant vectors (e.g., recombinant expression vectors) comprising same.

Abstract: Genes encoding bacterial oxygen binding proteins are provided. Recombinant expression of at least one of the present bacterial hemoglobin genes increased the growth characteristics and/or carotenoid production levels in microbial host cells grown under microaerobic conditions.

Abstract: A method for the in vivo bioconversion of cyclic carotenes having a ?-ionone ring to the corresponding aryl carotene is provided. Gram negative host cells expressing a heterologous, codon-optimized gene encoding a carotene desaturase are grown in the presence of a suitable cyclic carotene substrate to effect the production of aromatic carotenoids.

Abstract: The invention features a method of producing heterologous molecules in cells under the regulatory control of a metabolite and metabolic flux. The method can enhance the synthesis of heterologous polypeptides and metabolites.

Abstract: The present invention provides methods to engineer microorganisms for the production of C30-aldehyde carotenoids. Specifically, various combinations of crtM, sqs, crtN and crtN2 genes from Staphylococcus aureus and Methylomonas sp. 16a can be co-expressed in transformant hosts, leading to the production of diaponeurosporene monoaldehyde, diapocarotene monoaldehyde, and/or diapocarotene dialdehyde. In a preferred embodiment, the genetically engineered pathway is introduced into a strain of Escherichia coli that has been engineered for the expression of carotenoids, and the C30-carotenoid product is diapocarotene dialdehyde.

Abstract: Mutant phytoene desaturase genes are provided encoding polypeptides having the ability to convert a phytoene desaturase substrate to tetradehydrolycopene. Both in vivo and in vitro methods are provided using the present phytoene desaturases for tetradehydrolycopene production.

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 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: Genes have been isolated from Pectobacterium cypripedii encoding geranylgeranyl pyrophosphate (GGPP) synthase (CrtE), phytoene synthase (CrtB), phytoene desaturase (Crtl), lycopene cyclase (CrtY), ?-carotene hydroxylase (CrtZ), and zeaxanthin glucosyl transferase (CrtX) activity. The genes and their products are useful for the conversion of farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.

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 producing the high-value ketocarotenoid astaxanthin by the green microalga Chlorella zofingiensis in dark-heterotrophic cultures shows excellent growth and high-yield astaxanthin production on glucose-supplemented media in the dark. The specific growth rate and astaxanthin yield can be as high as 0.031 h?1, and 10.3 mg l?1, respectively, which are the highest so far reported in heterotrophic algal cultures. The light-independent astaxanthin-producing ability of Chlorella zofingiensis can be employed for commercial production of astaxanthin using industrial fermenters.

Abstract: Genes have been isolated from Rhodococcus and Deinococcus which encode a specific lycopene ?-cyclase capable of converting acyclic carotenoids with at least one ?-end group to the corresponding asymmetric carotenoid containing a single ?-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: Genes have been isolated from Methylomonas 16a sp. encoding the isoprenoid biosynthetic pathway. The genes and gene products are the first isolated from a Methylomonas strain that is capable of utilizing single carbon (C1) substrates as energy sources. The genes and gene products of the present invention may be used in a variety of ways for the production of isoprenoid compounds in a variety of organisms.

Abstract: The present invention relates to proteins which have an enzymatic activity for converting ?-carotene into zeaxanthin or canthaxanthin into astaxanthin, to nucleic acids which encode these proteins, to nucleic acid constructs comprising these nucleic acids, to genetically manipulated organisms where the genetic manipulation causes or increases the gene expression of this nucleic acid by comparison with a wild-type, and to processes for preparing xanthophyll derivatives.

Abstract: The present invention relates to a crystalline carotenoid compound, such as ?-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: 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: The invention relates to the development of a novel medium starting from fresh water using a salt solution complex comprising a non conventional salt, namely KCl and conventional salts such as NaCl and MgSO4 in the proportion defined in the specification to generate biomass of higher carotenogenesis in particular Betacarotene and its isomers using Dunaliella salina ARL5 in a single stage of active growth. D.salina ARL5, a local isolate, has an unique property of having a wide range of pH and temperature tolerance which is beneficial in production terms as it is grown in external conditions (i.e. outdoors).

Abstract: A unique carotenogenic biosynthetic gene cluster has been isolated from Panteoa agglomerans strain DC404, wherein the genetic organization of the cluster is crtE-idi-crtY-crtI-crtB-crtZ. The genes contained within this cluster encode geranylgeranyl pyrophosphate (GGPP) synthetase (CrtE), isopentenyl pyrophosphate isomerase (Idi), lycopene cyclase (CrtY), phytoene desaturase (CrtI), phytoene synthase (CrtB), and ?-carotene hydroxylase (CrtZ). The gene cluster, genes and their products are useful for the conversion of farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.

Abstract: The present invention relates to an isolated DNA sequence encoding an enzyme (e.g., farnesyl pyrophosphate synthase) in 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 astaxanthin and/or farnesyl pyrophosphate using such transformed host cells is also provided.

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: Genes have been isolated from strain DC260, a member of the Enterobacteriaceae family, encoding geranylgeranyl pyrophosphate (GGPP) synthetase (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 farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.

Abstract: A unique carotenogenic biosynthetic gene cluster has been isolated from Panteoa agglomerans strain DC404, wherein the genetic organization of the cluster is crtE-idi-crtY-crtI-crtB-crtZ. The genes contained within this cluster encode geranylgeranyl pyrophosphate (GGPP) synthetase (CrtE), isopentenyl pyrophosphate isomerase (Idi), lycopene cyclase (CrtY), phytoene desaturase (CrtI), phytoene synthase (CrtB), and &bgr;-carotene hydroxylase (CrtZ). The gene cluster, genes and their products are useful for the conversion of farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.

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: 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 production of carotenoid is accomplished using a DNA molecule that encodes a polypeptide as obtained from Haematococcus pluvialis, Phaffia rhodozyma, or Saccharomyces cerevisiae, having isopentonyl pyrophosphate (IPP) isomerase activity, or DNA molecule having a nucleotide sequence that hybridizes thereto. In particular, one can introduce such a DNA molecule into a carotenoid-producing microorganism, culture the microorganism thus transformed, and then obtain carotenoids in the culture broth and cells.

Abstract: A process for the conversion of (3R,3′R,6′R)-lutein to 3′-epilutein, a carotenoid precursor for industrial production of naturally occurring (3R,3′R)-zeaxanthin, is disclosed.

Abstract: Genes have been isolated from Pectobacterium cypripedii encoding geranylgeranyl pyrophosphate (GGPP) synthase (CrtE), phytoene synthase (CrtB), phytoene desaturase (Crtl), lycopene cyclase (CrtY), &bgr;-carotene hydroxylase (CrtZ), and zeaxanthin glucosyl transferase (CrtX) activity. The genes and their products are useful for the conversion of farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.

Abstract: The present invention relates to carotenoid overproducing bacteria The genes of the isoprenoid pathway in the bacterial hosts of the invention have been engineered such that certain genes are either up-regulated or down regulated resulting in the production of carotenoid compounds at a higher level than is found in the un-modified host. Genes that may be up-regulated include the dxs, idi, ispB, lytB and ygbBP genes. Additionally it has been found that a partial disruption of the yjeR gene has the effect of enhancing carotenoid production.

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: 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: The invention relates to methods for mapping and characterizing catalytic domains in enzymes, preferably plant enzymes and those enzymes within the carotene synthesis family and more specifically &egr;-cyclase enzymes regulating formation of &egr;,&egr;-carotene. The methods include reverse PCR and site-directed mutagenesis for generating chimera and truncations or site-directed mutations of enzymes, respectively. These chimera, truncations or site directed mutants of &egr;-cyclase enzymes are useful in the characterization of the sequence residues conferring catalytic domains for the enzymes, and more specifically, the identification of single residues regulating catalytic activity for enzymes that are important in plant growth and photosynthesis. Chimeric enzymes generated by the methods of the invention can also be used to create transgenci hosts which are augmentated in their expression of specific carotene products.

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 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: Method of production of &bgr;-carotene by fermentation in mixed culture using (+) and (−) strains of Blakeslea trispora

Abstract: Mutations in genes having no direct relationship to the carotenoid biosynthetic pathway have been found to increase carbon flux through that pathway. Complete disruption in the deaD, mreC, and yfhE genes were effective. Additionally where genes of the lower carotenoid pathway reside on a plasmid having either a p15A or pMB1 replicon, mutations in the thrS, rspA, rpoC, yjeR, and rhoL were found effective.

Abstract: There is disclosed herein a process for purifying a crude xanthophyll which is characterized by acting a lipase on the crude xanthophyll, which is obtained by extracting algae with a solvent, in the present of water under an acidic condition without adjusting the pH whereupon the contaminating neutral lipids (mono-, di- and triglycerides) are selectively hydrolyzed, subjecting the lipase-treated liquor to oil/water separation and removing free fatty acids from the oil layer fractioned thereby elevating the content of xanthophyll.

Abstract: This invention discloses a cyclic process for the production of astaxanthin rich biomass by cultivating Haematococcus algae. Modified nutrient medium containing 4 nitrogen sources is used for culturing the algae. Green motile cells produced are converted into dormant red cysts which are chilled and stressed for vigorous multiplication. Nutrients are added gradually and the initial germination is carried out without carbon dioxide sparging. The dilution stage is also effected in the absence of carbon dioxide. The stressed red cysts are regerminated and the cycle repeated to produce a biomass enriched with astaxanthin.

Abstract: Membranous bacteria that produce astaxanthin and other carotenoids are described, as well as isolated nucleic acids and expression vectors that can be used for producing carotenoids in microorganisms.

Abstract: The invention relates to a novel method for the production of &bgr;-carotene from submerged cultures of mucoral fungi such as Blakeslea, Choanephora or Phycomyces by adding lectin to the culture medium and performing pH control once fermentation has started. The method involves &bgr;-carotene recovery stage that makes it possible to simplify the process, optimize yields and increase purification of the product.

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: This invention provides a pigment-containing substance for feed additives consisting of a microorganism culture precipitate which contains a high concentration of carotenoid compounds. This pigment-containing substance for feed additives is resistant to oxygen, light and so on, and can be stably conserved for a long time period.

Abstract: The invention relates to coryneform bacteria which, instead of the singular copy of an open reading frame (ORF), gene or allele naturally present at the particular desired site (locus), have at least two copies of the open reading frame (ORF), gene or allele in question, preferably in tandem arrangement, and optionally at least a third copy of the open reading frame (ORF), gene or allele in question at a further gene site, and processes for the preparation of chemical compounds by fermentation of these bacteria.

Abstract: The present invention provides means and methods of transforming bacteria, fungi including yeast, animal and plant cells, seeds, tissues and whole plants in order to yield transformants capable of expressing &bgr;-carotene 15, 15′ dioxygenase and accumulating vitamin A aldehyde. The present invention further provides means and methods to biotechnically produce retinoids using cells, tissues, organs or whole organisms which natively or after transformation accumulate &bgr;-carotene or which take up &bgr;-carotene from the medium. The present invention also provides DNA molecules encoding &bgr;-carotene 15, 15′ dioxygenases derived from different sources and taxonomic groups of living organisms designed to be suitable for carrying out the method of the invention, and plasmids or vector systems comprising said molecules.

Abstract: The instant invention is drawn towards transformed strains of Fusarium sporotrichioides effective for the production of lycopene. The transformed strains comprise an expression cassette having three genes encoding, respectively, geranylgeranyl-pyrophosphate synthase, phytoene synthase and phytoene desaturase (i.e. Tri5crtE, Tri5crtB and Tri5crtl). The transformed strains of Fusarium sporotrichioides of the instant invention produce lycopene at levels of up to 0.5 milligrams per gram culture dry weight.

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: Described herein is a novel three domain gene from Schizochytrium, denoted carotene synthase, that encodes a protein with three different enzymatic activities: phytoene dehydrogenase (PD), phytoene synthase (PS), and lycopene cyclase (LC). Also described is the isolated gene encoding the carotene synthase, homologues thereof, the enzyme encoded by such gene, biologically active portions and homologues thereof, recombinant nucleic acid molecules, microorganisms and plants that have been genetically modified to increase or decrease the action of such gene, and methods of producing carotenoids and derivatives thereof or methods of producing microorganisms and lipid products lacking pigmentation using the knowledge of the carotene synthase described herein.