Abstract: Provided herein are biocatalysts and systems thereof for pterin-dependent enzymes and pathways and methods of making and using the same. Provided herein in some embodiments are biocatalysts having a pterin source and a pterin-dependent enzymatic pathway biologically coupled to the pterin source. Tetrahydrobiopterin (referred to herein as BH4 or BH 4) can be the pterin source. The BH4 can be synthesized by a tetrahydrobiopterin synthesis pathway. The tetrahydrobiopterin synthesis pathway can include a GTP cyclohydrase; a pyruvoyl tetrahydropterin synthase; a sepiapterin reductase, and/or any combination thereof. The biocatalyst can further contain a pterin-dependent enzymatic pathway. The pterin-dependent enzymatic pathway can be amino acid mono-oxygenase, phenylalanine hydroxylase, tryptophan hydroxylase, tyrosine hydroxylase, nitric oxide synthase, alkylglycerol monooxygenase, and/or any combination thereof.

Abstract: The invention provides an organism for expressing foreign DNA, the organism engineered to accept standard DNA carriers. The genome of the organism codes for intracytoplasmic membranes and features an interruption in at least one of the genes coding for restriction enzymes. Further provided is a system for producing biological materials comprising: selecting a vehicle to carry DNA which codes for the biological materials; determining sites on the vehicle's DNA sequence susceptible to restriction enzyme cleavage; choosing an organism to accept the vehicle based on that organism not acting upon at least one of said vehicle's sites; engineering said vehicle to contain said DNA; thereby creating a synthetic vector; and causing the synthetic vector to enter the organism so as cause expression of said DNA.

Abstract: The invention relates to processes of producing a fermentation product, comprising liquefying a starch containing material with an alpha-amylase; pre-saccharifying and/or saccharifying and fermenting using a fermentation organism in the presence of a carbohydrate source generating enzyme and a cellulolytic composition The invention also relates to methods of dewatering whole stillage.

Abstract: The present invention relates to isolated polypeptides having glucoamylase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Abstract: The present invention relates to processes for producing fermentation products from starch-containing material, wherein an alpha-amylase, a thermostable protease, and optionally a carbohydrate-source generating enzyme and/or pullulanase, are present and/or added during liquefaction. The invention also relates to compositions suitable for use in a process of the invention.

Abstract: Provided are isolated polypeptides having glucoamylase activity, catalytic domains, and polynucleotides encoding the polypeptides, catalytic domains. Also provided are nucleic acid constructs, vectors and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides, catalytic domains.

Abstract: The invention relates to processes and means for the biotechnologically fermentative production of riboflavin (hereinafter also referred to as vitamin B2) and means for the implementation of this process, in particular a modified microbial host cell with increased riboflavin yield. The invention thus provides new processes and means for the regulation of the expression of enzyme activities involved in the riboflavin production of the host cell.

Abstract: The present invention is directed to novel genes mediating the carbon catabolite repression (CCR) of gluconeogenic genes. Furthermore, the polypeptides encoded by said genes as well as the use of said genes in a process for the production of a target fermentation product is provided. Processes for generating such microorganisms are also provided by the present invention. The invention is also related to a genetically engineered microorganism and its use for the production of a target fermentation product, wherein the gluconeogenic genes are relieved from CCR within said microorganism.

Abstract: The present invention relates to newly identified mRNA stabilizing elements useful for the production of a target fermentation product, such as e.g. vitamins or enzymes, in particular riboflavin (vitamin B2), biotin, pantothenic acid (vitamin B5), folic acid, thiamin, pyridoxine (vitamin B6), vitamin B12, xylanase, amylase, protease, glucanase, amylomaltase or maltogenic amylase.

Abstract: The present invention relates to a improved process for the biotechnological production of compounds for which ribose-5-phosphate, ribulose-5-phosphate or xylulose-5-phosphate is biosynthetic precursor like riboflavin (vitamin B2), FAD, FMN, pyridoxal phosphate (vitamin B6), guanosine, GMP, adenosine, AMP. The invention further pertains to the generation of the organism producing those compounds. It furthermore relates to the generation of mutated transketolases that allow normal growth on glucose but reduced growth on gluconate when introduced into the production strains and to polynucleotides encoding them.

Abstract: The present invention relates to processes for producing fermentation products from starch-containing material, wherein liquefied mash is saccharified or pre-saccharified using a thermostable carbohydrate-source generating enzyme before fermentation or SSF.

Abstract: A truncated pullulanase variant of a parent pullulanase belonging to family GH57 comprising an X47 domain and the use thereof.

Abstract: The present invention relates to a method for producing a hydrolysate of from lignocellulose-containing material, comprising pre-treatment with low temperature, hydrolysis and fermentation, wherein hydrolysis is performed by contacting the lignocellulose-containing material with an enzyme composition comprising at least 10% xylanase enzyme protein w/w%.

Abstract: The present invention relates to a improved process for the biotechnological production of compounds for which ribose-5-phosphate, ribulose-5-phosphate or xylulose-5-phosphate is biosynthetic precursor like riboflavin (vitamin B2), FAD, FMN, pyridoxal phosphate (vitamin B6), guanosine, GMP, adenosine, AMP. The invention further pertains to the generation of the organism producing those compounds. It furthermore relates to the generation of mutated transketolases that allow normal growth on glucose but reduced growth on gluconate when introduced into the production strains and to polynucleotides encoding them.

Abstract: The present invention relates to isolated polypeptides having glucoamylase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Abstract: The present invention relates to isolated polypeptides having glucoamylase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Abstract: The invention relates to processes and means for the biotechnologically fermentative production of riboflavin (hereinafter also referred to as vitamin B2) and means for the implementation of this process, in particular a modified microbial host cell with increased riboflavin yield. The invention thus provides new processes and means for the regulation of the expression of enzyme activities involved in the riboflavin production of the host cell.

Abstract: The present invention provides an improved biotechnological production of riboflavin (also referred herein as vitamin B2) through modification in the operon containing the riboflavin biosynthetic genes (rib operon), in particular modifications of/in the leader sequences (rib leader) upstream of the corresponding riboflavin biosynthetic genes (rib operon). Furthermore, the present invention relates to genetically engineered microorganisms carrying said modified sequences, processes to generate said modified sequences/microorganisms and the use thereof for production of riboflavin.

Abstract: The present invention relates to newly identified mRNA stabilizing elements useful for the production of a target fermentation product, such as e.g. vitamins or enzymes, in particular riboflavin (vitamin B2), biotin, pantothenic acid (vitamin B5), folic acid, thiamin, pyridoxine (vitamin B6), vitamin B12, xylanase, amylase, protease, glucanase, amylomaltase or maltogenic amylase.

Abstract: The invention relates to a process for the purification of riboflavin comprising the steps of (a) precipitating a first crystalline form of riboflavin, (b) isolating the first crystalline form of riboflavin, (c) transforming the first crystalline form of riboflavin into a second crystalline form of riboflavin under conditions that decompose diluted DNA, and (d) isolating the second crystalline form of riboflavin, provided that at ambient temperature the first crystalline form of riboflavin is thermodynamically less stable than the second crystalline form of riboflavin.

Abstract: The present invention relates to a improved process for the biotechnological production of compounds for which ribose-5-phosphate, ribulose-5-phosphate or xylulose-5-phosphate is biosynthetic precursor like riboflavin (vitamin B2), FAD, FMN, pyridoxal phosphate (vitamin B6), guanosine, GMP, adenosine, AMP. The invention further pertains to the generation of the organism producing those compounds. It furthermore relates to the generation of mutated transketolases that allow normal growth on glucose but reduced growth on gluconate when introduced into the production strains and to polynucleotides encoding them.

Abstract: A titanylphthalocyanine crystal having an X-ray diffraction spectrum having plural diffraction peaks and a primary particle diameter not greater than 0.2 ?m, wherein a maximum diffraction peak is observed at a Bragg (2?) angle of 27.2±0.2°; main peaks are observed at 9.4°, 9.6° and 24.0°; and a minimum diffraction peak is observed at 7.3°; and preferably no diffraction peak is observed at an angle greater than 7.3° and less than 9.4° when a specific X-ray of CuK? having a wavelength of 1.542 ? irradiates the titanylphthalocyanine crystal.

Abstract: A process for the preparation of a target fermentation product comprising cultivation of an aerobic microorganism to produce such product, wherein in the microorganism the efficiency of the respiratory chain is increased, transformed and novel nucleotide sequences.

Abstract: Riboflavin-producing Bacillus subtilis which is resistant to proline analogue, and a method for producing riboflavin using the Bacillus subtilis are provided.

Abstract: Riboflavin-producing Bacillus subtilis which is resistant to threonine analogue, and a method for producing riboflavin using the Bacillus subtilis are provided. The subject Bacillus subtilis referred to as Bacillus subtilis CJKB0002 has the accession number KCCM-10446, having been deposited in Korean Culture Center of Microorganisms (KCCM), 361-221, Yurim B/D, Hongie-1-dong, Seodaemun-gut, Seoul 120-091 Republic of Korea, on Nov. 18, 2002.

Abstract: The invention provides a process of producing bio-available folate, i.e. folic acid having an increased portion of monoglutamyl folate and a decreased proportion of polyglutamyl folate, by culturing food-grade micro-organisms containing an active heterologous or homologous polyglutamyl hydrolase activity or containing increased activities of folate biosynthesis enzymes. The genes encoding the polyglutamyl hydrolase and the folate biosynthesis enzymes may be of various origin, e.g. from rodents or other mammals including man. Also provided is a foodstuff, especially a dairy product containing such monoglutamyl folate-producing micro-organisms.

Abstract: Riboflavin-producing Bacillus subtilis which is resistant to proline analogue, and a method for producing riboflavin using the Bacillus subtilis are provided.

Abstract: Riboflavin-producing Bacillus subtilis which is resistant to threonine analogue, and a method for producing riboflavin using the Bacillus subtilis are provided.

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 a transformed cell which is transformed by at least one gene of enzymes participating in biosynthesis of tetrahydrobiopterin and a process for the production of a biopterin compound using the same. In accordance with the present invention, the biopterin compound can be produced in large quantities in an industrial advantageous manner from less expensive materials.

Abstract: Vectors and recombinant bacteria for overproducing riboflavin, in which nucleic acid overproducing riboflavin biosynthetic proteins is introduced in the chromosome of the host organism, e.g. at multiple sites and in multiple copies per site. A rib operon having at least five genes is used to make such recombinant bacteria.

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 invention relates to a process for producing corynebacteria comprising one or more modified genomic sequences, where a vector is used which does not replicate in corynebacteria and whose nucleic acid is not recognized by corynebacteria as foreign.

Abstract: This invention is directed to the transformation of the flavinogenic yeasts, Pichia guilliermondii and Candida famata, and mutants thereof, by electroporation (electrotransformation) and by spheroplast transformation. The invention is also directed to nucleic acid constructs such as vectors, plasmids, and ARS sequences which transform flavinogenic yeasts, and mutants thereof, at a high level and in a stable manner so as to result in stably transformed yeast host cells which express/produce recombinant products. This invention also is directed to flavinogenic yeasts, Pichia guilliermondii and Candida famata, and mutants and temperature sensitive mutants thereof, which produce or overproduce riboflavin.

Abstract: Vectors and recombinant bacteria for overproducing riboflavin, in which nucleic acid overproducing riboflavin biosynthetic proteins is introduced in the chromosome of the host organism, e.g. at multiple sites and in multiple copies per site. A rib operon having at least five genes is used to make such recombinant bacteria.

Abstract: Provided are a monoclonal antibody making it possible to detect a native fibrin monomer, which is produced at the initial state of blood coagulation, and soluble fibrin; a hybridoma; and an immunoassay for detecting the initial stage of blood coagulation with high sensitivity, quickly, using the monoclonal antibody. Using a fibrinogen analog in blood as an immune source, cell fusion is carried out to prepare a monoclonal antibody which is not reactive with fibrinogen and is specifically and simultaneously reactive with a native fibrin monomer (that is, a fibrin monomer which is present in a body fluid, in particular in blood, and is not solubilized) and soluble fibrin. The fibrin monomer analog is preferably fibrinogen treated with bathroxobin, which is a snake venom.

Abstract: The present invention provides a recombinant bacterium for the over-production of riboflavin. The recombinant bacterium has has been transformed by three or four vectors, two of which each comprise either a DNA sequence coding for the riboflavin synthesizing enzymatic activities of Bacillus subtilis or a DNA sequence which is substantially homologous and one or more transcription elements and a third and/or fourth vector comprising either a DNA sequence coding for the ribA gene product of Bacillus subtilis or a DNA sequence which is substantially homologous and optionally a transcription element whereby one or a plurality of copies of each of these vectors has/have been integrated at three or four different sites within the bacterium's chromosome.

Abstract: An industrially more advantageous method for the production of metabolites biologically synthesized via phosphoribosyl pyrophosphate (PRPP) is provided, making use of metabolically modified strains in which transketolase activity is deficient or reduced in comparison with the parent strain.

Abstract: A process for increasing the riboflavin content in spray-dried discharges from riboflavin fermentations comprises inducing the lysis of the cells contained in the fermentation discharge by specific production or activation of enzymes which break down the cell walls and proteins. This autolysis can be induced in the riboflavin-producing strains of Ashbya gossypii, Eremothecium ashbyii and other microorganisms by incubation at from 35 to 55.degree. C. with substantial exclusion of oxygen for from 6 to 28 hours.

Abstract: Vectors and recombinant bacteria for overproducing riboflavin, in which nucleic acid overproducing riboflavin biosynthetic proteins is introduced in the chromosome of the host organism, e.g. at multiple sites and in multiple copies per site. A rib operon having at least five genes is used to make such recombinant bacteria.

Abstract: Vectors and recombinant bacteria for overproducing riboflavin, in which nucleic acid overproducing riboflavin biosynthetic proteins is introduced in the chromosome of the host organism, e.g. at multiple sites and in multiple copies per site. A rib operon having at least five genes is used to make such recombinant bacteria.

Abstract: The present invention relates to the genes for riboflavin biosynthesis in the fungus Ashbya gossypii and to genetic engineering processes for preparing riboflavin using these genes and gene products.

Abstract: The present invention provides a process for producing riboflavin efficiently at a low cost, wherein riboflavin is formed and accumulated in a medium by culturing a microorganism carrying a recombinant DNA prepared by inserting into a vector DNA a DNA which is derived from a microorganism belonging to the genus Corynebacterium or Brevibacterium and which, upon introduction into a riboflavin-requiring microorganism, confers on the microorganism the ability to complement its riboflavin requirement.

Abstract: A substantially pure recombinant human myeloperoxidase heme-containing precursor, comprising a glycoprotein of 84 KD with the amino acid sequence coded for by the nucleotide sequence from 145 to 2235 corresponding to codons 49 to 745 in phase after the first methionine codon in FIG. 1 produced by culturing prokaryotic or eukaryotic cells transformed by a vector for the expression of human myeloperoxidase precursor in said cells.

Abstract: Genetically engineered E. coli carry vectors containing inserts that code for Clostridium histolyticum collagenase. These inserts code for: (a) a form of collagenase having a molecular weight of about 68,000 daltons in the essential absence of larger forms of collagenase; (b) the 68 kd form of collagenase and a fusion polypeptide consisting of the collagenase protein fused to at least a portion of the .beta.-galactosidase protein of E. coli or (3) the 68 kd form of collagenase and polypeptides of molecular weight of from above about 68,000 daltons to about 100,000 daltons and having the enzymatic activity of C. histolyticum collagenase as indicated by digestion of .sup.3 H-acetylated collagen and by specific inhibition by 1,10-phenanthroline plus EDTA. The collagenase genes in the transformed E. coli are expressed efficiently in the transformed cells to yield enzymatically active and immunologically cross-reactive collagenase.

Abstract: The present invention provides a process for producing riboflavin by fermentation, a method for providing microorganisms having an improved riboflavin-producing capability, and strains of microorganisms having improved riboflavin-producing ability. The strains of the present invention belong to the genus Bacillus, have reduced activity of hydrolysing phosphoric acid from 5'-guanylic acid, and have the ability of producing riboflavin. Mutants used in the processes of this invention have an improved ability of producing riboflavin and are capable of producing or accumulating a large amount of riboflavin in the culture medium. The processes of this invention are therefore suitable for producing riboflavin in an effective manner at a low cost.

Abstract: A method of systematically generating and isolating highly flavinogenic strains of Candida famata. The strains Candida famata GA18Y8-6#2 dgr and Candida famata GA18Y8-6#2#11 have been developed and produce yields of ca. 7.0 to 7.5 grams per liter per 6 days. The method includes a combination of iterative mutagenizing steps and protoplast fusion steps performed on the parent strain and the descendant strains which are selected following each step according to a screening protocol.

Abstract: A method of purifying ferment-produced riboflavin, wherein the impure riboflavin is suspended in water or dilute aqueous acid, and the suspension is heated at a temperature of from 75.degree. to 130.degree. C. for from 0.3 to 3 hours with stirring, during which time crystal transformation takes place, after which the mixture is cooled and the purified product is isolated in known manner.

Abstract: Riboflavin is removed from fermentation suspensions by them being heated at from 50.degree. to 90.degree. C. for from 1 to 3 hours, than cooled to from 0.degree. to 30.degree. C. over a period of from 1 to 5 hours, and subsequently being centrifuged to give a sediment fraction and liquid fraction in such a way that the sediment fraction contains predominantly riboflavin crystals as solid, and the liquid fraction contains virtually no crystalline riboflavin, and, where appropriate, resuspending the sediment fraction in from 0.5 to 2 parts by volume of water per part by volume of sediment fraction and repeating procedure c.

Abstract: Strains of yeast of the species Candida famata are disclosed which can produce 10 grams of riboflavin per liter in 6 days, and in particular, strains identified by ATCC Accession Nos. 20849 and 20850. Riboflavin yields of more than 20 grams per liter in 200 hours have been achieved. Strains of the present invention have increased sensitivity to iron inhibition of flavinogenesis and have enhanced riboflavin production per cell at increased iron concentrations in the fermentation medium. The invention also is directed toward a process for selecting improved microorganisms which are resistant to inhibition of growth by depleted medium. Such selected mircoorganisms are then tested for riboflavin overproduction. The present invention is also directed toward a selection process in which mutated microorganisms are cultured in the presence of tubercidin, a purine analog. Mutant strains resistant to tubercidin are then tested for riboflavin over-production.