Abstract: A gene encoding an enzyme required for operation of a novel biochemical pathway for oxidation of the reduced phosphorus (P) compound phosphite was cloned from Pseudomonas and also found in other bacteria. The enzyme (designated PtxD) was overproduced in the host Escherichia coli by use of a recombinant system and purified to homogeneity via a two-step affinity protocol and characterized. The enzyme stoichiometrically produces NADH and phosphate from NAD and phosphite. Mechanistic studies indicate stereoselective transfer of hydride from phosphite to the Re-face of NAD+ with observed steady-state kinetic isotope effects of 2.1 on Vmax and 1.8 on Vmax/Km. The novel enzyme is useful for methods requiring regenerating the cofactor NADH, for use in synthetic oxidoreductases, and to synthesize chiral compounds, complex carbohydrates, and isotopically-labelled compounds.

Abstract: A glyoxal-guanosine-group compound is prepared either by reacting glyoxal-guanine with any one of ribose-1-phosphate and 2-deoxyribose-1-phosphate in the presence of purine nucleoside phosphorylase, or by reacting glyoxal-guanine with any one selected from the group consisting of uridine, 2′-deoxyuridine and thymidine, together with phosphate ion, in the presence of purine nucleoside phosphorylase and pyrimidine nucleoside phosphorylase. The glyoxal-guanosine-group compound is then decomposed by alkali, whereby a guanosine-group compound consisting of guanosine and 2′-deoxyguanosine is prepared.

Abstract: A glyoxal-guanosine-group compound is prepared either by reacting glyoxal-guanine with any one of ribose-1-phosphate and 2-deoxyribose-1-phosphate in the presence of purine nucleoside phosphorylase, or by reacting glyoxal-guanine with any one selected from the group consisting of uridine, 2′-deoxyuridine and thymidine, together with phosphate ion, in the presence of purine nucleoside phosphorylase and pyrimidine nucleoside phosphorylase. The glyoxal-guanosine-group compound is then decomposed by alkali, whereby a guanosine-group compound consisting of guanosine and 2′-deoxyguanosine is prepared.

Abstract: According to the present invention, UDP-N-acetylgalactosamine and an N-acetylgalactosamine-containing carbohydrate can be produced using a protein having UDP-N-acetylglucosamine 4-epimerase activity.

Abstract: The present invention relates to a process for producing 5′-inosinic acid or 5′-guanylic acid for use in seasonings or the like from inosine or guanosine or a precursor thereof using an adenosine triphosphate (ATP)-regenerating microorganism containing a DNA encoding a protein that has the activity of forming 5′-inosinic acid or 5′-guanylic acid from inosine or guanosine. Further, the present invention relates to a novel protein having the inosine-guanosine kinase activity, a gene encoding said protein, a recombinant DNA containing said gene, and a microorganism which is transformed with said recombinant DNA.

Abstract: The present invention relates to the production of biomass for the isolation of ccc plasmid DNA comprising culturing a bacterial transformant in a bioreactor containing an antibiotic-free batch medium under batch-conditions and, at the end of the batch phase, feeding under feed-back conditions the portion of a feed-back medium after the rise of the concentration of dissolved oxigen above a threshold-set point. Said feed-back medium comprises besides a carbon source a magnesium salt, preferably in concentrations above 20 mM. Preferably, the bacterial transformant is harvested after the end of the culture and frozen or freeze-dried. Also preferred is that ccc plasmid DNA is, optionally directly, isolated after harvesting the bacteria.

Abstract: The present invention provides a method of nucleic acid, including DNA, immunization of a host, including humans, against disease caused by infection by a strain of Chlamydia, specifically C. pneumoniae, employing a vector, containing a nucleotide sequence encoding a CPN100605 polypeptide of a strain of Chlamydia pneumoniae and a promoter to effect expression of the CPN100605 polypeptide in the host. Modifications are possible within the scope of this invention.

Abstract: The invention provides a method for producing a cytosine nucleoside compound from pentose-1-phosphate and cytosine or a derivative thereof using a nucleoside phosphorylase reactive to cytosine or a bacterium having the enzyme activity. The invention also provides a method for specifically reducing an activity to degrade the substrates or the product, resulting in efficient production of the cytosine nucleoside compound. According to the invention, little by-product is produced in producing cytonucleocide compounds.

Abstract: A glyoxal-guanosine-group compound is prepared either by reacting glyoxal-guanine with any one of ribose-1-phosphate and 2-deoxyribose-1-phosphate in the presence of purine nucleoside phosphorylase, or by reacting glyoxal-guanine with any one selected from the group consisting of uridine, 2′-deoxyuridine and thymidine, together with phosphate ion, in the presence of purine nucleoside phosphorylase and pyrimidine nucleoside phosphorylase. The glyoxal-guanosine-group compound is then decomposed by alkali, whereby a guanosine-group compound consisting of guanosine and 2′-deoxyguanosine is prepared.

Abstract: The present invention is directed methods of purine synthesis using a modified amidotransferase, wherein the modified glutamine PRPP amidotransferase has reduced sensitivity to end-product inhibition of synthesizing purine nucleotides.

Abstract: This invention provides methods for practical enzymatic conversion of GDP-mannose to GDP-fucose. These methods are useful for efficient synthesis of reactants used in the synthesis of fucosylated oligosaccharides.

Abstract: A method for preparing saccharide compositions is disclosed. The method is reiterative and comprises the following three steps.

Abstract: The present invention is directed to a modified amidotransferase. The amino acid sequence of the modified amidotransferase differs from the amino acid sequence of a native E. coli glutamine PRPP amidotransferase in that one or more amino acid residues of the modified amidotransferase are substituted at positions equivalent to amino acid positions in Bacillus amidotransferase selected from the group consisting of 282, 283, 307, and 347 of SEQ ID NO:1, wherein the modified amidotransferase is less sensitive to end-product inhibition than is the native E. coli glutamine PRPP amidotransferase.

Abstract: The present invention is directed to isolated polynucleotides coding for phosphoglucose isomerase (pgi) from coryneform bacteria. In addition, the invention includes methods for increasing the metabolic flux through pentose phosphate cycle of bacteria by reducing or eliminating the activity of pgi. These methods may be used to increase the fermentative production of nucleotides, vitamins and amino acids.

Abstract: The invention provides a GDP-mannose-pyrophosphorylase, which is suitable for use in continuous multiple stage processes. The mannose- or mannose-derivative-specific GDP-mannose-prophosphorylase, which can be isolated from microorganisms, has a specific activeity of≧2U/mg, is prepared.

Abstract: The invention provides methods for producing products comprising improved host cells genetically engineered to have uncoupled productive and catabolic pathways. In particular, the present invention provides host cells having a modification in nucleic acid encoding an endogenous enzymatic activity that phosphorylates D-glucose at its 6th carbon and/or a modification of nucleic acid encoding an enzymatic activity that phosphorylates D-gluconate at its 6th carbon. Such improved host cells are used for the production of products, such as, ascorbic acid intermediates. Methods for making and using the improved host cells are provided. Nucleic acid and amino acid sequences for glucokinase and gluconokinase are provided.

Abstract: The present invention relates to an ATP-regeneration reaction system comprising the steps of acting adenylate kinase on AMP to convert it into ADP and acting a polyphosphoric acid synthetase in the presence of a polyphosphoric acid compound to convert it into ATP and a polyphosphoric acid compound; an ATP-regeneration reaction system comprising the steps of acting a phosphotransferase on AMP in the presence of a polyphosphoric acid compound to convert it into ADP and then acting a polyphosphoric acid synthetase on the resulting ADP to convert it into ATP; and a method for detecting or inspecting adenine nucleotide or RNA and an ATP-amplification method, which make use of the foregoing regeneration system.

Abstract: The present invention provides methods directed to detecting antibodies that specifically bind to a varicella zoster polypeptide, detecting the presence of a varicella zoster virus in an animal, diagnosing a disease caused by varicella zoster virus, and detecting a varicella zoster virus having a single nucleotide polymorphism in ORF68. The present invention also provides a vaccine composition, a method for producing a modified attenuated varicella zoster virus, isolated polynucleotides, and isolated polypeptides, and viruses.

Abstract: Propargylethoxyamino nucleosides are disclosed having the structure wherein R1 and R2 are —H, lower alkyl, or label; B is a 7-deazapurine, purine, or pyrimidine nucleoside base; W1 is —H or —OH; W2 is —OH or a moiety which renders the nucleoside incapable of forming a phosphodiester bond at the 3′-position; and W3 is —PO4, —P2O7, —P3O10, phosphate analog, or —OH. Additionally, a primer extension method is provided employing the above propargylethoxyamino nucleosides.

Abstract: Disclosed are purified and isolated DNA sequences encoding eukaryotic proteins possessing biological properties of inosine 5′-monophosphate dehydrogenase (“IMPDH”). Illustratively, mammalian (e.g., human) IMPDH-encoding DNA sequences are useful in transformation or transfection of host cells for the large scale recombinant production of the enzymatically active expression products and/or products (e.g., GMP) resulting from IMPDH catalyzed synthesis in cells. Vectors including IMPDH-encoding DNA sequences are useful in gene amplification procedures. Recombinant proteins and synthetic peptides provided by the invention are useful as immunological reagents and in the preparation of antibodies (including polyclonal and monoclonal antibodies) for quantitative detection of IMPDH.

Abstract: In a method for producing a target substance utilizing a microorganism comprising culturing the microorganism in a medium to produce and accumulate the target substance in the medium and collecting the target substance, there is used, as the microorganism, a mutant strain or a genetic recombinant strain constructed from a parent strain of the microorganism having a respiratory chain pathway of high energy efficiency and a respiratory chain pathway of low energy efficiency as respiratory chain pathways, and having either one or both of the following characteristics:

Abstract: Long chain nucleic acids such as plasmids are separated from contaminants including protein, RNA, DNA and mixtures thereof by using hydrophobic interaction chromatography or hydrophobic interaction chromatography and ion exchange chromatography. To carry out hydrophobic interaction chromatography, first and second columns containing first and second column materials are used. The first column material adsorbs protein and RNA at a salt concentration at which plasmid is not adsorbed to produce an eluate containing plasmid and DNA. The second column material adsorbs plasmid and DNA from the eluate at a salt concentration which the first column material adsorbs protein and RNA but not the plasmid. The plasmid is eluted from the second column material at a salt concentration at which plasmid is eluted. A tandem column may be formed by connecting in series the first and second columns. The connection between the columns is broken before eluting plasmid from the second column material.

Abstract: Nucleoside 5′-phosphate ester is produced by culturing a bacterium belonging to the genus Escherichia having an ability to produce nucleoside 5′-phosphate ester, in which ushA gene and aphA gene do not function normally, in a medium to produce and accumulate nucleoside 5′-phosphate ester in the medium, and collecting the nucleoside 5′-phosphate ester from the medium.

Abstract: The present invention discloses nucleic acid enzymes capable of cleaving single-stranded DNA in a site specific manner.

Abstract: This invention relates to a process for producing a sugar nucleotide, in which a) a culture broth of a microorganism capable of producing NTP from a nucleotide precursor, or a treated product of the culture broth, and b) a culture broth of a microorganism capable of producing a sugar nucleotide from a sugar and NTP, or a treated product of the culture broth, are used as enzyme sources; a process for producing a complex carbohydrate, in which the above-described a) and b) and c) a culture broth of a microorganism, an animal cell or an insect cell capable of producing a complex carbohydrate from a sugar nucleotide and a complex carbohydrate precursor, or a treated product of the culture broth, are used as enzyme sources; a process for producing a complex carbohydrate, in which a culture broth of a microorganism, an animal cell or an insect cell capable of producing a complex carbohydrate from a sugar nucleotide and a complex carbohydrate precursor, or a treated product of the culture broth, is as an enzyme sour

Abstract: 2′-deoxy-2′-alkylnucleotides useful for stabilizing enzymatic nucleic acid molecules and antisense molecules.

Abstract: This invention relates to a process for producing a sugar nucleotide, in which a) a culture broth of a microorganism capable of producing NTP from a nucleotide precursor, or a treated product of the culture broth, and b) a culture broth of a microorganism capable of producing a sugar nucleotide from a sugar and NTP, or a treated product of the culture broth, are used as enzyme sources; a process for producing a complex carbohydrate, in which the above-described a) and b) and c) a culture broth of a microorganism, an animal cell or an insect cell capable of producing a complex carbohydrate from a sugar nucleotide and a complex carbohydrate precursor, or a treated product of the culture broth, are used as enzyme sources; a process for producing a complex carbohydrate, in which a culture broth of a microorganism, an animal cell or an insect cell capable of producing a complex carbohydrate from a sugar nucleotide and a complex carbohydrate precursor, or a treated product of the culture broth, is as an enzyme sour

Abstract: Subject of this invention is a process for the chemical synthesis of polynucleotides having totally or partially random sequences, based on the utilization, as synthesis monomer units for the random sequence part, or presynthesized mononucleotides and dinucleotides. Said synthesis is carried out on separate supports, so that on each of those supports is alternated at least one reaction cycle wherein a mixture of said dinucleotides is bound, with at least one reaction cycle wherein a mononucleotide is bound, and that in a preferred embodiment at the end of the n cycles required for a codon synthesis, the supports are mixed and then redivided into one or more reaction containers. The resulting polynucleotides are such that, for the random sequence part, each trinucleotide unit is fit to match only a limited number of codons, predefined for each unity in number and sequence, and the genetic code degeneracy effects can thus be eliminated. FIG.

Abstract: There is provided a method for convenient judgement of a possibility of the onset of leukemia of an ovine individual caused by the bovine leukemia virus (BLV) by means of genetic engineering techniques. A method for judging a possibility of the onset of ovine leukemia caused by bovine leukemia virus (BLV), wherein an ovine individual, in which an amino acid sequence defined by the amino acid numbers of 70 and 71 of the &bgr;1 domain of the ovine MHC Class II DR&bgr; chain is Ser-Arg or Gln-Thr, is judged to have a risk of the onset of the leukemia, or an ovine individual, in which the amino acid sequence is Arg-Lys, is judged to have resistance to the onset of the leukemia.

Abstract: The present invention is directed to a DNA element that enhances the translation of the human amyloid precursor protein (APP) gene. The enhancer may be incorporated into expression vectors to enhance recombinant protein production. In addition, the invention is directed to an assay that utilizes vectors containing the translation enhancer element for the purpose of identifying agents that modulate the expression of the human amyloid precursor protein. These agents will ultimately be used to suppress APP expression in patients with Alzheimer's disease.

Abstract: The present invention is directed to a modified glutamine PRPP amidotransferase and a method of using that modified enzyme to enhance the biosynthesis of purine nucleotides. The modified glutamine PRPP amidotransferase has at least one amino acid of the allosteric A sites or the catalytic C sites of said amidotransferase substituted with a non-native amino acid, wherein the substitution reduces the sensitivity of the enzyme to end product inhibition relative to the native glutamine PRPP amidotransferase enzyme.

Abstract: A process for producing uridine diphosphate-N-acetylglucosamine (UDPAG) from uridylic acid (UMP) and N-acetylglucosamine by use of microorganism cells, characterized by adding N-acetylglucosamine kinase thereto. According to the present invention, UDPAG can be efficiently produced even when N-acetylglucosamine is used as a substrate.

Abstract: A nucleic acid substance is efficiently produced by cultivating a microorganism whose repressor protein for purine operon does not function in a normal manner, preferably a strain whose gene encoding the repressor protein on a chromosome (purR) is disrupted, in a culture medium so that the nucleic acid substance should accumulate in the culture medium, and collecting the substance from the culture medium.

Abstract: The invention discloses a two-step process for recovery of thuringiensin, comprising adsorbing the thuringiensin from fermentation broth by calcium silicate, and dissociating the thuringiensin by dibasic sodium phosphate. The resulting thuringiensin can be further purified by using semi-preparative HPLC and electrodialysis to remove the excess salts from the recovered thuringiensin solution.

Abstract: Propargylethoxyamino nucleosides are disclosed having the structure wherein R1 and R2 are —H, lower alkyl, or label; B is a 7-deazapurine, purine, or pyrimidine nucleoside base; W1 is —H or —OH; W2 is —OH or a moiety which renders the nucleoside incapable of forming a phosphodiester bond at the 3′-position; and W3 is —PO4, —P2O7, —P3O10, phosphate analog, or —OH. Additionaly, a primer extension method is provided employing the above propargylethoxyamino nucleosides.

Abstract: This invention is an integrated instrument for the high-capacity electrophoretic analysis of biopolymer samples. It comprises a specialized high-voltage, electrophoretic module in which the migration lanes are formed between a bottom plate and a plurality of etched grooves in a top plate, the module permitting concurrent separation of 80 or more separate samples. In thermal contact with the bottom plate is a thermal control module incorporating a plurality of Peltier heat transfer devices for the control of temperature and gradients in the electrophoretic medium. Fragments are detected by a transmission imaging spectrograph which simultaneously spatially focuses and spectrally resolves the detection region of all the migration lanes. The spectrograph comprises a transmission dispersion element and a CCD array to detect signals. Signal analysis comprises the steps of noise filtering, comparison in a configuration space with signal prototypes, and selection of the best prototype.

Abstract: A method for the stepwise creation of phosphodiester bonds between desired nucleosides resulting in the synthesis of polynucleotides having a predetermined nucleotide sequence by preparing an initiation substrate containing a free and unmodified 3′-hydroxyl group; attaching a mononucleotide selected according to the order of the predetermined nucleotide sequence to the 3′-hydroxyl of the initiating substrate in a solution containing a catalytic amount of an enzyme capable of catalyzing the 5′ to 3′ phosphodiester linkage of the 5′-phosphate of the mononucleotide to the 3′-hydroxyl of the initiating substrate, wherein the mononucleotide contains a protected 3′-hydroxyl group, whereby the protected mononucleotide is covalently linked to the initiating substrate and further additions are hindered by the 3′-hydroxyl protecting group. Methods in which a mononucleotide immobilized on a solid support is added to a free polynucleotide chain are also disclosed.

Abstract: A nucleoside/tide compound having the structure NUC-L-S-LB/LG is described wherein NUC is a nucleoside/tide having a nucleobase portion B, L is a rigid linkage, S is a spacer; and LB/LG is a member of a linkage pair or a label. NUC is attached to L through B such that when B is a purine, L is attached to the 8-position of the purine, when B is 7-deazapurine, L is attached to the 7-position of the 7-deazapurine, and when B is pyrimidine, L is attached to the 5-position of the pyrimidine. In an important aspect of the invention, L has the structure wherein each of n, o and p are integers ranging from 0 to 3, and the sum of n, o and p is at least 2, and each of W, X, Y and Z is selected from the group consisting of carbon and nitrogen. The invention further includes polynucleotide compounds comprising the nucleoside/tide, and primer extension methods utilizing the nucleoside/tide, particularly when used in combination with certain mutant polymerase enzymes.

Abstract: The invention relates to a compound of formula (I) ##STR1## Wherein: B is adenine, guanine or hypoxanthineZ is hydrogen or a negative chargeR is --[CH.sub.2 CH(R.sup.1)--O].sub.n -R.sup.2, --CH.sub.2 CH.sub.2 X, in whichR.sup.1 is hydrogen or (C.sub.1 -.sub.6) alkylR.sup.2 is hydrogen or (C.sub.1 -.sub.6) alkyln is a number from 1 to 6X is OH, F, NR.sup.3 R.sup.4R.sup.3 and R.sup.4 are independently from each other hydrogen or (C.sub.1 -.sub.6) alkyland to methods of enzymatically treating these compounds with biocatalysts having cyclic phosphodiesterase activity.

Abstract: There is disclosed a process for producing a nucleoside derivative of the formula [I]: ##STR1## characterized by contacting 2',3',5'-O-triacylribonucleoside derivative of the formula [II]: ##STR2## with an ester hydrolase: (i) capable of regio-selectively deacylating the acyl group at 5'-O-position in the formula [II] above, and(ii) having an amino acid sequence encoded by a gene which hybridizes to a nucleotide sequence encoding an amino acid sequence of SEQ ID NO:1.

Abstract: The invention relates to a multifunctional nucleoside didesoxyribosyl or nucleoside deoxyribosyl transferase which has one or more of the following additional activities (desoxy) nucleoside kinase, nucleoside reductase desaminase, or DNA polymerase activity. Utilizing the multifunctional enzyme results in a variety of nucleic acid products. These products can be prepared using sequential reactions in a single batch process wherein the sequential reaction can be caused to occur by varying process conditions in a manner which turns on or off the requisite activities causing the sequential reactions to occur. An example of a product prepared in this manner is dideoxyribofuranoside triphosphate. Certain of the resultant products have pharmaceutical activities, e.g. antiviral agents. Lactobacillus leichmannii (DSM 20076) is a source of the multifunctional nucleoside deoxyribosyl transferase.

Abstract: Nucleic acids encoding various proteases, from a mammal, reagents related thereto, including specific antibodies, and purified proteins are described. Methods of using said reagents and related diagnostic kits are also provided.

Abstract: An oligonucleotide analog or an antisense molecule, which is minimally hydrolyzable with an enzyme in vivo, has a high sense strand binding ability, and is easily synthesized, is provided. It is an oligo- or polynucleotide analog containing one or more monomer units being nucleotide analogs of the general formula: ##STR1## where B may be identical or different, and is a pyrimidine or purine nucleic acid base, or a derivative thereof.

Abstract: A process for preparing a sugar nucleotide from a nucleotide by using a yeast cell, characterized in that both a nucleoside diphosphate-sugar pyrophosphorylase and a sugar 1-phosphate are present in the reaction system. According to this process, various sugar nucleotides, which have been prepared only in low productivity by the conventional yeast cell process, can be efficiently prepared.

Abstract: A class of 4,7-dichlororhodamine compounds useful as fluorescent dyes are disclosed having the structure ##STR1## wherein R.sub.1 -R.sub.6 are hydrogen, fluorine, chlorine, lower alkyl, lower alkene, lower alkyne, sulfonate, sulfone, amino, amido, nitrile, lower alkoxy, linking group, or combinations thereof, or, when taken together, R.sub.1 and R.sub.6 is benzo, or, when taken together, R.sub.4 and R.sub.5 is benzo; Y.sub.1 -Y.sub.4 are hydrogen or lower alkyl, or, when taken together, Y.sub.1 and R.sub.2, Y.sub.2 and R.sub.1 Y.sub.3 and R.sub.3, or Y.sub.4 and R.sub.4 is propano, ethano, or substituted forms thereof, and X.sub.1 -X.sub.3 taken separately are selected from the group consisting of hydrogen, chlorine, fluorine, lower alkyl, carboxylate, sulfonic acid, --CH.sub.2 OH, and linking group. In another aspect, the invention includes reagents labeled with the 4,7-dichlororhodamine dye compounds, including deoxynucleotides, dideoxynucleotides, and polynucleotides.

Abstract: The present invention relates to a process for producing nucleoside 5'-triphosphates (NTP) from nucleoside 5'-diphosphates (NDP) other than adenosine 5'-diphosphate (ADP), characterized by using a polyphosphate kinase as an enzyme and polyphosphate as the phosphate donor; and application of this process to various glycosylation reactions.This process makes it possible to conveniently and economically synthesize NTP from NDP enzymatically. Also, it becomes possible thereby to economically recycle and synthesize sugar nucleotides and synthesize, for example, oligosaccharides associating therewith without resort to expensive phosphoenol pyruvate, ATP, etc. in the reactions for reproducing or converting NDP into NTP in the systems for enzymatically synthesizing oligosaccharides by combining, for example, with the synthesis of sugar nucleotides.

Abstract: The present invention relates to a process for preparing sugar nucleotides from nucleotides and sugar derivatives by the use of yeast, characterized in that the reactions are conducted at 20.degree. C. or below. According to this process, even when the preparation is conducted on an enlarged scale, a reduction in the yield of a sugar nucleotide can be inhibited by a very simple means; lowering the reaction temperature 20.degree. C. or below. Thus, the process is an extremely practical one applicable to the mass-production of sugar nucleotides.

Abstract: This invention relates to a process for the production of fructose 2,6-bisphosphate which comprises effecting reactions among (i) fructose 6-phosphate, (ii) glucose, (iii) fructose or (iv) glucose 6-phosphate, ATP and a phosphate donor, in the presence of (i) fructose 6-phosphate 2-kinase (PFK 2) and an enzyme which converts ADP into ATP (ADP/ATP converting enzyme), (ii) PFK 2, an ADP/ATP converting enzyme, hexokinase or glucokinase and glucose 6-phosphate isomerase, (iii) PFK 2, an ADP/ATP converting enzyme and hexokinase or glucokinase, or (iv) PFK 2, an ADP/ATP converting enzyme and glucose 6-phosphate isomerase; to a process for the production of fructose 2,6-bisphosphate which comprises allowing diesterase to coexist in a solution containing fructose 1,2-cyclic, 6-bisphosphate; and to a process for the purification of fructose 2,6-bisphosphate which comprises adding zinc salt to a solution containing fructose 2,6-bisphosphate, removing formed precipitate of impurities and adding a zinc salt to the result

Abstract: A method is provided using centrifugation to prepare a seal of solidified wax, grease or polymer mix over an aqueous reagent in a reaction container such that the reagent is separated from contact with the atmosphere. The amount of solidified wax, grease or polymer mix is not sufficient when melted to a liquid to separate the reagent from contact with the atmosphere under gravity. A reagent and solidified wax, grease or polymer mix are combined in a container. During centrifugation and heating, the solidified wax, grease or polymer mix melts to a liquid, and centrifuging causes the liquid to form over the reagent a layer that completely separates the reagent from the atmosphere. As centrifugation continues, the liquid is cooled and solidified to form the seal. Additional reagents are preferably added on top of the seal such that when the container is heated and the seal melted the upper and lower reagents mix for reaction.

Abstract: This invention relates to a fermentation process for high-yield production of plasmid DNA in E coli strains. In the disclosed process, a slow growth rate of cells is controlled and maintained by an automated nutrient feed scheme based on dissolved oxygen concentration (DOC) and pH. This controlled slow growth rate promotes high plasmid DNA stability during host cell replication. As a result, high yield production of plasmid DNA is achieved.