79 results about "Flavin adenine dinucleotide" patented technology

Object: An object of the present invention is to provide a more practically advantageous enzyme usable as a reagent for measuring blood glucose than the known enzymes used as blood glucose sensors.

Method for Achieving the Object: A modified flavin adenine dinucleotide dependent glucose dehydrogenase (FADGDH) with more improved heat stability than FADGDH derived from wild-type FADGDH, the modified FADGDH being derived from preferably a eukaryote, more preferably a filamentous fungus, and furthermore preferably an Aspergillus fungus, and, for example, those having a primary structure with at least one amino acid substituted, deleted, inserted or added to FADGDH having an amino acid sequence represented by SEQ ID Nos. 2 or 46 in the sequence table.

The invention discloses a recombinant human NADH (nicotinamide-adenine dinucleotid) dehydrogenase subunit-4 gene and a constructing method of an expression vector thereof. The nucleotide sequence of the gene is shown in SEQ ID NO:1, and the size of the nucleotide sequence is 2889 bp. The constructing steps of adeno-associate virus vectors are as follows: firstly constructing a recombinant adeno-associated virus vector containing the human NADH dehydrogenase subunit-4 gene, and then coating, infecting, purifying, concentrating and identifying the recombinant adeno-associated virus. According to the method, the recombinant adeno-associated virus vector with the recombinant human NADH dehydrogenase subunit-4 gene can be quickly and simply constructed, and is packaged to obtain the adeno-associated virus vector with complex defects. The gene can be used for gene therapy of LHON (leber's hereditary optic neuropathy).

The invention relates generally to the field of industrial microbiology and alcohol production. More specifically, the invention relates to the use of thiamine, biosynthetic precursors of thiamine, nicotinic acid, nicotinamid, nicotinic acid riboside, nicotinamid riboside, or other biosynthetic precursors of nicotine adenine dinucleotide (NAD) to improve butanol production. Butanol production can be improved by providing sufficient amounts of thiamine, biosynthetic precursors of thiamine, nicotinic acid, nicotinamid, nicotinic acid riboside, nicotinamid riboside, or other biosynthetic precursors of nicotine adenine dinucleotide (NAD) in the production media.

Provided is an enzyme that is further advantageous in terms of practical aspects when compared to publicly known enzymes for blood sugar sensors, and that can be used in a blood sugar level measuring reagent.

A flavin adenine dinucleotide-dependent glucose dehydrogenase that has amino acid sequence including a specific amino acid in an amino acid sequence shown in SEQ ID NO: 2 or an amino acid sequence that has a 60% homology therewith, and that has an improved temperature dependency.

This disclosure provides a method of determining the viability of at least one cell via quantification of nicotinamide adenine dinucleotide (phosphate) (NAD(P)H), flavin adenine dinucleotide (FAD), and/or collagen. The method includes the steps of contacting the at least one cell with light having a wavelength of from 700 to 900 nm, using two photon excitation, or from 335 to 400 nm, using one photon excitation, to induce an optical response from the NAD(P)H, FAD, and/or collagen and measuring the optical response. The method also includes the step of quantifying one or more of an amount, spatial localization, and/or time-dependent response of the NAD(P)H, FAD, and/or collagen utilizing the optical response.

The invention discloses a biosynthetic gene cluster of ikarugamycin and an application of the biosynthetic gene cluster. The nucleotide sequence of the biosynthetic gene cluster of the ikarugamycin is as shown in the 3411th to 15678th sites of SEQ ID NO. 1 and contains three genes, namely a heterozygous polyketone/non-ribosomal polypeptide synthetase gene ikaA, FAD (Flavin Adenine Dinucleotide) dependent oxidoreductase gene ikaB and similar Michael cyclization reaction enzyme gene ikaC. All the genes and protein information relevant to the biosynthesis of the ikarugamycin are helpful to interpret the biosynthesis mechanism of natural products of a polycyclic tetramate large-ring lactam family so as to provide a theoretical basis and materials for further genetic modification. The genes and proteins can be used for digging or creating compounds or genes and proteins applicable to the medicine and health industry and industry and agriculture in nature.

The invention provides a carbon dioxide measurement kit. The carbon dioxide measurement kit mainly comprises the following components: 50 to 200mM of a buffer solution, 10 to 50% of a stabilizer, 0.1 to 5% of a surface active agent, 0.1 to 5g/L of a preservative, 1 to 20mM of a reaction accelerator, 2 to 10mM of 3-acetylpyridine adenine dinucleotide (reduction type), 5 to 50g/L of phosphoenolpyruvic acid, 50 to 500U/L of phosphoenolpyruvate carboxylase, and 1 to 3KU/L of malate dehydrogenase; a recycling system of 3-acetylpyridine adenine dinucleotide (reduction type) comprises (1) 2 to 20g/L of glucose substrate, (2) 100 to 2,000U/L of corresponding glucose dehydrogenase, and (3) 1 to 5mM of 3-acetylpyridine adenine dinucleotide. The carbon dioxide measurement kit is convenient to use and simple to operate; the components participating in coupling reaction are extra added, so that no pollution caused by endogenous and allogenic substances are introduced; the rate of the recycling system is below the main reaction rate, thus the main reaction cannot be interfered; the carbon dioxide measurement kit is high in stability and can be used for a long time.

The invention relates to the field of a biological detection instrument, and particularly relates to a biosensor based on glucose dehydrogenase. The biosensor is composed of an insulation substrate, an electrode system, a detecting reagent, a middle interlayer, a covering layer, a reaction room and a sample injection channel; the reaction room is provided with air holes. Enzyme in the detection reagent is flavin adenine dinucleotide dependent form glucose dehydrogenase; the electronic medium is commonly formed by ruthenium complex and 2, 6-dichlorophen indophenol. The invention further provides a detecting method of the biosensor. The biosensor solves the problem that the pure ruthenium complex is very hard to accept electronic from the flavin adenine dinucleotide dependent form glucose dehydrogenase, thus the biosensor has the advantages of good reaction sensitivity and linearity.

The object is to provide a novel enzyme which enables to determine the glucose level more accurately, a bacterium capable of producing the enzyme, and use of the enzyme. Disclosed is a flavin adenine dinucleotide-binding glucose dehydrogenase having the following properties (1) to (3):(1) the enzyme has an activity of catalyzing a reaction for oxidizing a hydroxyl group in glucose in the presence of an electron acceptor to produce glucono-delta-lactone; (2) the enzyme has a molecular weight of about 100 kDa as measured by SDS-polyacrylamide gel electrophoresis or about 400 kDa as measured by gel filtration chromatography; and (3) the enzyme is less reactive to maltose, D-fructose, D-mannose and D-galactose. Also disclosed is a microorganism Aspergillus oryzae which can produce the enzyme. Further disclosed are a glucose determination method, a reagent for the determination of glucose, and a kit for the determination of glucose, each utilizing the enzyme.

The method of detecting thalassemia by optical analysis of blood components is a spectral detection method that is based on the fluorescence spectra of a set of biomolecules, including tyrosine, tryptophan, nicotinamide adenine dinucleotide, and flavin adenine dinucleotide, which are all found in blood plasma, and porphyrin, which is found in red blood cells (RBCs). Measured ratios of intensity maxima between tryptophan and nicotinamide adenine dinucleotide, flavin adenine dinucleotide and nicotinamide adenine dinucleotide, tyrosine and tryptophan, and the normal form of porphyrin and the basic form of porphyrin may each be used, alone or in combination, to diagnose a patient as suffering from thalassemia.

The invention provides a triglyceride determination kit. The kit comprises a liquid single reagent R1, the reagent R1 includes the following components with concentration: lipoprotein esterase, glycerol kinase, glycerol phosphate oxidase, peroxidase, piperazine-1,4-diethanesulfonic acid, NaOH, 4-aminoantipyrine, adenosine triphosphate, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methylaniline sodium orN-ethyl-N-(2-hydroxy-3-propyl)-3'5-dimethoxyaniline sodium salt, sodium glutamate, magnesium acetate, flavin adenine dinucleotide, polyethylene glycol, and a betaine solution. The kit belongs to the technical field of biological detection, and the triglyceride determination kit provided by the invention significantly enhances the anti-interference ability while improving the stability of the reagent, and has good measurement accuracy for triglyceride.

The invention relates to a cholesterol oxidase affinity matrix formed by linking a cholesterol oxidase affinity ligand and a chromatography media; more preferably, the cholesterol oxidase affinity ligand is FAD (Flavin Adenine Dinucleotide) segmental riboflavin and the chromatography media is Sepharose, chitosan or polyvinyl benzene; the invention further provides a synthesis method for cholesterol oxidase affinity matrix, and a large-scale purification method for cholesterol oxidase by using the cholesterol oxidase affinity matrix; the cholesterol oxidase can be rapidly separated and purified in large scale by using the cholesterol oxidase affinity matrix provided by the invention; on the condition of only using one step of affinity chromatography, the pure cholesterol oxidase is obtained; the consumption time of the entire purification step is short, the active recovery rate of protein is higher and the large-scale popularization and application is applicable.

The present invention provides a novel L-glutamate oxidase, a gene encoding the enzyme, and a method for producing the enzyme. By use of a gene encoding the enzyme, L-glutamate oxidase can be readily prepared at low costs through a recombinant DNA technique. The novel L-glutamate oxidase has the following physicochemical properties:

• • (A) action: catalyzing the following reaction:

L-glutamic acid+O₂+H₂O→α-ketoglutaric acid+H₂O₂+NH₃;

(B) substrate specificity: being specific to L-glutamic acid;

(C) molecular weight and subunit structure: molecular weight as determined through SDS-polyacrylamide gel electrophoresis of 70,000±6,000, molecular weight as determined through gel filtration of 140,000±10,000, and being a dimer formed of the same subunits having a molecular weight of 70,000±6,000;

(D) optimum pH: around pH 6.0 to 8.5;

(E) heat stability: being stable up to 60° C. at a pH of 7.4 for 30 minutes; and

(F) coenzyme: flavin adenine dinucleotide (FAD).

The invention relates to a FAD-Na2 freeze-dried acanthopanax powder spasmolytic preparation and a method for preparing the same. The FAD-Na2 freeze-dried acanthopanax powder spasmolytic preparation is a drug composition made by mixing FAD-Na2 serving as medicinal active ingredients and pharmaceutically acceptable excipients. The preparation method is as follows: the FAD-Na2 is used as raw material and added with certain spices of excipients according to certain proportions; the mixture is prepared and developed by a technical means provided by the invention into freeze-dried acanthopanax powder spasmolytic agent which can be used for subcutaneous injection, intramuscular and intravenous injection and used to cure a variety of diseases caused by lacks of vitamin B2.

The invention discloses a production and purification method of glucose dehydrogenase by taking FAD (Flavin Adenine Dinucleotide) as a prothetic group, and belongs to the field of bioengineering. Theproduction and purification method is characterized by expressing the glucose dehydrogenase sourced from burkholderia cepacia by taking Escherichia coli as a host, carrying out batch feeding, controlling the temperature in stages, and expressing the glucose dehydrogenase. According to the production and purification method disclosed by the invention, the activity of the glucose dehydrogenase is upto 22200.0U/L, and the mycelia content is up to 69.8g/L; a crude enzyme solution is separated and purified through three-step chromatography, so that recombinase of which the specific enzyme activityis 10<9>U/mg, the property problem of enzyme is solved, and the recombinase is suitable for industrial production.