64 results about "Menadione" patented technology

The invention provides a method for biosynthesizing menadione by using yarrowia lipolytica. The method is realized by constructing recombinant yarrowia lipolytica, and the construction method of the recombinant yarrowia lipolytica for synthesizing menadione comprises the following steps: intensively expressing a mevalonic acid pathway in the yarrowia lipolytica, and synthesizing menadione by introducing a synthesis pathway of heterologous geranyl and geranyl pyrophosphate. A sufficient isoprene side chain donor is provided for biosynthesis of menadione; on the basis, an aromatic isopentenyl transferase coding gene which is optimized by codons and has menadione substrate specificity is over-expressed in yarrowia lipolytica. The construction method of the yarrowia lipolytica for biosynthesis of menadione provided by the invention is simple to operate, the constructed yarrowia lipolytica can obtain a menadione product with relatively high concentration by taking commercially available menadione as a substrate, the biosynthesis steps are relatively short, and the yarrowia lipolytica has relatively high production and application values.

The invention provides a 2-succinyl-5-enol acetonyl-6-hydroxy-3-cyclohexene-1-carboxylic acid (SEPHCHC) synthase mutant, which is characterized in that on the basis of a bacillus subtilis SEPHCHC synthase gene MenD, 12 SEPHCHC synthase mutants are obtained by virtue of a site-specific mutagenesis PCR (Polymerase Chain Reaction) technology, and the SEPHCHC synthase gene MenD is used as a base. Recombinant bacteria of bacillus subtilis or bacillus natto or bacillus licheniformis or bacillus amyloliquefaciens for expressing the mutant are respectively constructed, and the yield of heptaene menadione fermented by the recombinant bacteria is 74%-203% of the yield of heptaene menadione fermented by the recombinant bacteria of bacillus subtilis for expressing wild type SEPHCHC synthase MenD.

The invention relates to a method for synthesizing menadione through in-tank indirect electrooxidation of 2-methylnaphthalene. Metal titanium is used as a substrate, a Sn-Ir-doped titanium-based PbO2 electrode is prepared by adopting a coating method and is used as an electrolytic anode, and a Pb electrode is used as a cathode. 2-methylnaphthalene, a solvent, cobaltous sulfate, sulfuric acid, ammonium sulfate, an electrocatalyst and water are prepared into anolyte according to a certain proportion, a sulfuric acid aqueous solution serves as catholyte, an oil-water two-phase reaction solution is obtained from an anode in a two-phase electrolysis mode, an oil phase is directly refined to obtain menadione, and a water phase is reused after being refined. The method is short in technological process, few in three wastes, mild in condition and extremely low in cost, and has extremely high industrialization prospects.

The application provides a kind of aromatic isopentenyl transferase mutant, which is based on the aromatic isopentenyl transferase gene MenA derived from Bacillus subtilis ( Bacillus subtilis ) and six aromatic isopentenyl transferase mutants MenA-1 to MenA-6 are obtained by site-directed mutagenesis PCR technology, and the recombinant bacteria Bacillus subtilis expressing the above mutants are constructed, respectively. The yield of heptamethylnaphthoquinone fermented by the recombinant bacteria is 64.49 mg / L, 69.30 mg / L, 83.86 mg / L, 105.84 mg / L, 119.23 mg / L and 125.45 mg / L, respectively, which is 133%, 143%, 173%, 219%, 246% and 259% of the yield of heptamethylnaphthoquinone fermented by the Bacillus subtilis expressing the wild-type aromatic isopentenyl transferase MenA.

The invention discloses a novel crystal form of menadione and a preparation method, and relates to the technical field of organic chemistry. The novel crystal form comprises a crystal form T, and the crystal form T has diffraction peaks when the 2theta value is 9.970 + / -0.2 degrees, 12.250 + / -0.2 degrees, 14.978 + / -0.2 degrees and 19.141 + / -0.2 degrees. The preparation method of the novel crystal form comprises the following steps: dissolving a menadione crude product in a mixed solvent composed of an ester solvent and an alcohol solvent, cooling to 10-20 DEG C, and crystallizing to obtain the menadione crystal form T; the menadione crystal form T obtained through the method is a novel crystal form, the crystal form is good in stability and solubility, a solvent system is environmentally friendly, solvent residues are very small, the risk is low, the medication safety is better guaranteed, the defects of an existing crystal form can be overcome, and high economic benefits and social benefits are achieved.

The invention discloses a filamentous fungus genetic transformation system taking an endogenous KatG1 gene as a selection marker, a method and application. The method comprises the following steps: carrying out linear DNA fragment construction on an endogenous Pgpd promoter, an endogenous KatG1 gene, a target promoter and a target gene of filamentous fungi, then transforming the constructed linear DNA fragment into filamentous fungi cells, and finally screening through a menadione-containing culture medium to obtain the directionally improved filamentous fungi strain containing the target gene. According to the invention, the endogenous KatG1 gene of the fungus is used as a selection marker gene and menadione is used as screening resistance to construct a novel selection marker based on an endogenous gene and a genetic transformation system without exogenous DNA (Deoxyribose Nucleic Acid) such as a carrier and the like, so that the method has the advantages of universality, high efficiency and safety; an environment-friendly solution is provided for directional genetic improvement of filamentous fungi, and the method has important commercial application potential in directional breeding of fungus strains in the fields of agriculture, industry and the like.

The invention relates to a 2-succinyl-5-enol acetonyl-6-hydroxy-3-cyclohexene-1-carboxylic acid synthase mutant and application thereof, and belongs to the technical field of enzyme engineering. According to the invention, a key residue of 2-succinyl-5-enolacetonyl-6-hydroxy-3-cyclohexene-1-carboxylate synthase is obtained by combining protein sequence comparison and alanine scanning strategies, the key residue is subjected to random mutation, and the menadione pathway resistant intermediate metabolite 1, 2, 3, 4-tetracarboxylic acid synthase is obtained through multiple rounds of screening. The mutant MenDW322IR323F which is subjected to feedback inhibition by using 2, 4-dihydroxy-2-naphthoic acid is synthesized, the heptaene menadione production capacity of the recombinant bacillus subtilis constructed according to the method is improved, the yield is 1.22 times that of a control strain, and the final yield reaches 186.5 mg / L. The invention not only provides a new direction for transformation of a menadione approach of bacillus subtilis, but also is beneficial to industrial expanded production of heptaene menadione.

The present application discloses compositions comprising nanoparticles of vitamin K2, and their methods of use.

The invention provides application of a composition containing butyric acid producing bacteria and vitamin K2 in preparation of a medicine for treating osteoporosis, and relates to the technical field of biological medicines, the composition comprises butyric acid producing bacteria and vitamin K2; wherein the butyric acid producing bacteria are selected from any one or more of clostridium butyricum, rosella and clostridium praeparatum; and the vitamin K2 is selected from any one or more of menadione-4 or menadione-7. The composition can play a synergistic effect through an intestine-bone axis, and compared with a single component, the composition can more remarkably improve the bone mineral density. The composition has comprehensive effects and high safety, provides a brand-new nutrition intervention strategy with a synergistic effect for prevention and treatment of osteoporosis, and provides a brand-new possibility for preparation of medicines related to osteoporosis.

The invention discloses a method for improving the synthesis yield of vitamin K3, which comprises the following steps: S1, adding beta-menadione and sodium pyrosulfite into a container, adding ethanol as a solvent, stirring to react for 1-2 hours at the reaction temperature of 30-50 DEG C, heating to 40-70 DEG C, keeping the temperature for 10-60 minutes, and filtering while hot after the reaction is completed to obtain filtrate; and S2, adding sodium pyrosulfite into the filtrate, and then freezing, crystallizing, filtering, washing and drying to obtain a vitamin K3 finished product. When the vitamin K3 is prepared through cooling crystallization, the sodium pyrosulfite is added into the filtrate to be crystallized, so that the crystallization yield of the vitamin K3 can be effectively improved.

The invention relates to the technical field of menadione intermediate 1 preparation, and discloses a menadione intermediate 1 preparation method, which comprises: 1, introducing dicyclopentadiene into a first continuous flow reactor, and carrying out a depolymerization reaction; step 2, synchronously introducing cyclopentadiene in the step 1 and a solution containing menadione and a Lewis acid catalyst into a second continuous flow reactor; step 3, introducing the reaction liquid in the step 2 and water into a third continuous flow reactor for continuous quenching and liquid separation, and separating an organic phase; 4, drying the organic phase, concentrating, pulping with methanol, and purifying to obtain the menadione intermediate 1; the continuous flow reactor is adopted to realize continuous quenching and continuous liquid separation, the operation risk is reduced while the operation is simplified, the production safety is improved, the Lewis acid is adopted as the catalyst, the reaction process is mild in condition, small in acid consumption, short in reaction time and small in corrosion to equipment, and the three-waste pollution is greatly reduced.

The invention discloses a method for preparing menadione and heptaene menadione by utilizing supercritical fluid chromatography. The method for preparing the menadione and the heptaene menadione by using the supercritical fluid chromatography comprises the following steps: dissolving a sample containing the menadione and the heptaene menadione by using a dissolving reagent to prepare a test solution; carrying out supercritical fluid chromatographic separation on the test solution to respectively prepare the menadione and the heptaene menadione; the supercritical fluid chromatographic separation conditions are as follows: a mobile phase is a mixture of supercritical CO2 and an alkane solvent; the stationary phase comprises silica gel. The provided method for preparing menadione and heptaene menadione by using supercritical fluid chromatography realizes the separation of heptaene menadione and heptaene menadione, can reduce the use of organic reagents, and has the advantages of short separation time and good separation effect.

The present application discloses compositions comprising nanoparticles of vitamin K2, and their methods of use.

The present application relates to a kind of preparation methods of sodium bisulfite menadione, which is prepared by the reaction of 2-methyl-1,4-naphthoquinone and sodium bisulfite in the presence of K2O-MgO / Al2O3 as an aid in the mixed solution of organic solvent and water.The present application makes the conversion rate of raw materials >99% by introducing K2O-MgO / Al2O3 as an aid, the selectivity is greater than 95%, and can reduce the phenol, quinone impurities (content <800ppm) in the product.The process is simple, the reaction condition is mild, the process is stable, the purity of product is more than 98%, and the total yield is more than 94%.

The invention provides methods and compositions for preventing or treating (e.g., slowing the progression of, arresting, and / or reversing) tissue calcification in a subject in need thereof and, more particularly, the invention relates to methods of using menaquinone-7 (MK-7) and / or menaquinol-7 (MKH2-7) for preventing or treating (e.g., slowing the progression of, arresting, and / or reversing) tissue calcification in a subject with diabetes, chronic kidney disease, end stage renal failure, or a subject undergoing hemodialysis and / or receiving anticoagulant therapy. The invention further provides methods and compositions for reducing one or more symptoms of chronic obstructive pulmonary disorder (COPD), including using menaquinone-7 (MK-7) and / or menaquinol-7 (MKH2-7), for preventing or treating (e.g., slowing the progression of, arresting, and / or reversing) one or more symptoms of COPD.

The present invention relates to the use of high-dose vitamin K2, particularly menaquinone-7 (MK-7), in the treatment of coronary heart disease in high-risk subjects with a coronary artery calcification (CAC) score of ≧400 (e.g., at the start of treatment), and the treatment slows down the progression of coronary artery calcification.

This invention discloses a desmethylmenoprenone methyltransferase mutant, its construction method, and its applications, belonging to the field of biotechnology. Desmethylmenoprenone methyltransferase is a key enzyme in the MK-7 biosynthetic pathway, responsible for catalyzing the methylation reaction of desmethylmenoprenone to generate a series of methylnaphthoquinone (MK) products, ultimately forming MK-7. This invention, through metabolic network regulation and rational design, successfully obtained 19 single-point mutants, among which 5 mutants showed significantly higher catalytic efficiency than the wild-type enzyme. Experimental results show that the recombinant engineered bacteria constructed based on these mutants exhibit significant advantages in the whole-cell catalytic production of vitamin K2 (MK-7), with MK-7 yield increased by more than 20% compared to the wild-type strain. The excellent catalytic performance of the menG mutant provided by this invention provides strong technical support for the industrial production of vitamin K2, and has broad application prospects and market value.

The invention discloses a method for accelerating the color development speed of filamentous fungus resazurin and application of the method, and relates to the technical field of drug sensitivity detection. The invention provides a method for accelerating the color development speed of filamentous fungus resazurin, which comprises the following steps: S1, diluting a filamentous fungus bacterial liquid by using an MUELLER-HINTON culture medium, and adding the diluted filamentous fungus bacterial liquid into a to-be-detected drug hole for incubation; s2, performing suction filtration after incubation; s3, adding a color developing agent containing a catalyst for color development; s4, result interpretation; the catalyst in the step S3 is menadione. According to the MUELLER-HINTON culture medium, in the Aspergillus drug sensitivity detection process based on resazurin color development, after a catalyst is added, color development is accelerated, meanwhile, all medicine adding holes cannot be red, and the MIC value can be accurately obtained.

This application discloses, in part, isolated, stable and bioactive pegylated menaquinol derivatives for treating various diseases, and methods of using them. The present invention relates to bioactive menaquinol derivatives, compositions and formulations, and combinations thereof, their reduced and bioactive forms of menaquinol and salts thereof for treating vitamin K-related diseases including osteoporosis and osteopenia. In one embodiment, this application discloses novel biologically effective menaquinol derivatives (including their salts) such as menaquinol-7 derivatives and menaquinol-9 derivatives, and methods of using them for treating various diseases.

The present invention relates to novel animal feed supplements that comprise menaquinone-n, wherein (n) is the number of repeating 5 carbon units. The supplements further comprise saccharides, carotenoids, xanthophylls, minerals and electrolytes. The animal feed supplement comprises menaquinone-n, wherein (n) is the number of repeating 5 carbon units, enhances and the growth, reduces the mortality, and improves the feed conversion ratio, improves liver and bone health, and replaces the need for use of probiotics in livestock and / or poultry, especially monogastric livestock and / or poultry receiving such animal feed supplements.

The invention is applicable to the technical field of feeds, and provides a special nutrient enrichment premix for boars and a preparation method thereof. The compound feed additive is prepared by compounding beta-carotene, 25-hydroxyvitamin D3, dl-alpha-tocopheryl acetate, menadione sodium hydrogen sulfite, ammonium sulfate nitrate, riboflavin, pyridoxine hydrochloride, cyanocobalamin, nicotinamide, D-calcium pantothenate, folic acid, D-biotin, L-ascorbic acid, DL-methionine, zinc methionine chelate, selenium yeast, chromium picolinate, L-carnitine and glucose according to a specific proportion. A dynamic batching and high-precision mixing process is adopted for preparation. Through the synergistic effect of various nutrients, the sow milk quality can be remarkably improved, the birth weight of piglets is increased by 50 g or above, the survival rate is increased by 6-8% or above, the weaning weight is increased by 500 g or above, meanwhile, the sow estrus rate is increased by 5% or above, the problems that sows are poor in body condition, low in reproductive performance, low in piglet survival rate and the like in existing boar breeding are effectively solved, and extremely high breeding application value is achieved.

Compositions comprising nanoparticles of vitamin K2 and methods of use thereof are disclosed.

There are described compositions comprising an effective amount of a combination of two or more components, said components selected from acacetin, ACTI peptide, alpha-lipolic acid, alprostadil, anisomycin, apigenin, ascorbic acid, astragalus, berberine, β-lapachone, β-hydroxy-beta-methyl-butyrate, Bacopa monnieri, catechin, catechol, chamomile, chrysin, coumestrol, curcumin, dinitrophenol, dinoprost, ellagic acid, (−)-epigallocatechin gallate, green tea extract, fisetin, genistein, ginsenoside RE, glabridin, 18-α-glycyrrhetinic acid, 18-β-glycyrrhetinic acid, glycyrrhizin, hydroquinone, isoquercitrin (EMIQ), kaempferol, kuromanin, leucine, lithium, luteolin, luteolin, luteolinidin, melatonin, menadione, 1-methylnicotinamide (MNA), methyl salicylate, myricetin, nadide, niacin (vitamin B3), nicotinamide (NAM), nicotinamide mononucleotide (NMN), nicotinamide riboside (NR), nicotinic acid adenine dinucleotide (NaAD), nicotinic acid mononucleotide (NaMN), parsley (Petroselinium crispum), phenylephrine, pokeweed mitogen, 15-Δ prostaglandin J2, puromycin, quercetin, quinolinic acid, retinoic acid, trichostatin A, troxrutin, rutin, tryptophan, vitamin D3, withaferin A, wortmannin and zinc (including salts thereof).

This disclosure provides a method for alleviating aluminum toxicity in hydroponic peas based on menadione-mediated reactive oxygen species (ROS) metabolism. The core components include: regulating the concentration and duration of menadione treatment to optimize the spatiotemporal accumulation pattern of ROS (superoxide anion, hydrogen peroxide) in the roots of hydroponic peas; adjusting the dynamic response of endogenous antioxidant enzyme systems (SOD, POD, APX) to enhance menadione-mediated ROS scavenging capacity; controlling the initial ROS burst based on changes in plasma membrane NADPH oxidase (Rboh) gene expression and enzyme activity to inhibit the aluminum stress oxidative signaling cascade; and adjusting the timing and frequency of menadione administration based on subcellular-scale mitochondrial and chloroplast ROS leakage rates to alleviate organ-specific oxidative damage. This approach addresses five key issues: aluminum-induced oxidative damage, cellular redox imbalance, oxidative signaling cascade, organ-specific oxidative damage, and persistent aluminum toxicity metabolic disorders.

The invention provides methods and compositions for preventing or treating (e.g., slowing the progression of, arresting, and / or reversing) tissue calcification in a subject in need thereof and, more particularly, the invention relates to methods of using menaquinone-7 (MK-7) and / or menaquinol-7 (MKH2-7) for preventing or treating (e.g., slowing the progression of, arresting, and / or reversing) tissue calcification in a subject with diabetes, chronic kidney disease, end stage renal failure, or a subject undergoing hemodialysis and / or receiving anticoagulant therapy. The invention further provides methods and compositions for reducing one or more symptoms of chronic obstructive pulmonary disorder (COPD), including using menaquinone-7 (MK-7) and / or menaquinol-7 (MKH2-7), for preventing or treating (e.g., slowing the progression of, arresting, and / or reversing) one or more symptoms of COPD.

The invention relates to a hair growth medicine and application thereof in preparation of a medicine for preventing and treating male hormone alopecia, and belongs to the technical field of biological medicines. The invention discloses an application of menadione in preparation of a hair growth medicament. According to the present invention, the drugs capable of effectively treating male hormone alopecia are screened by using the high throughput screening method, comprise menthone, menadione and caffeic acid, and can significantly increase the hair follicle number, the hair follicle length and the corium layer thickness of the skin, the composition is used for treating male hormone alopecia, effectively promotes hair growth, and has no obvious side effects, and the expression quantity of the hair growth promoting gene is increased, and the expression quantity of the hair growth inhibiting gene is reduced.

The application provides a kind of aromatic isopentenyl transferase mutant, to be derived from Bacillus subtilis ( Bacillus subtilis )SEPHCHC synthase gene MenD is based, six aromatic isopentenyl transferase mutants MenD-1, MenD-2, MenD-3, MenD-4, MenD-5 and MenD-6 are obtained by site-directed mutagenesis PCR technology, and the recombinant bacteria Bacillus subtilis expressing the above mutants is respectively constructed, and the yield of heptamethylnaphthoquinone fermentation is 70.77 mg / L, 75.48 mg / L, 81.72 mg / L, 88.41 mg / L, 99.88 mg / L and 133.40 mg / L respectively, which is 116%, 123%, 134%, 144%, 162% and 218% of the yield of heptamethylnaphthoquinone of Bacillus subtilis expressing wild-type SEPHCHC synthase MenD respectively.

The invention discloses a method for preparing beta-menadione by using a compound emulsifier, which comprises the following steps: step 1, uniformly mixing high-molecular-weight alkanol and high-molecular alkyl sodium sulfonate to obtain the compound emulsifier; step 2, mixing beta-methylnaphthalene, water and a compound emulsifier to form an emulsion; step 3, mixing sodium dichromate with chromic anhydride, adding the mixture into water, and adding sulfuric acid to adjust the concentration of hexavalent chromium so as to form an oxidation solution; and 4, dropwise adding the oxidation liquid into the emulsion, carrying out a curing reaction on the reacted material, filtering, washing, and carrying out centrifugal dehydration to obtain the beta-menadione product. The invention has the following beneficial effects: the emulsion can be uniformly dispersed, the stability is better, the reaction speed with materials is high, the temperature range of reaction control is wider, and the side reaction is less; meanwhile, the compound emulsifier is better in stability and basically does not react with the oxidizing agent in the later period, the reaction process is more stable and continuous, and the obtained beta-menadione material is high in purity.

The present application relates to 2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylic acid synthase mutant and its application, and belongs to the technical field of enzyme engineering. The present application obtains key residues of 2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylic acid synthase by combining protein sequence comparison and alanine scanning strategy, randomly mutates the key residues, and obtains a mutant MenD which is resistant to feedback inhibition of 1,4-dihydroxy-2-naphthalene carboxylic acid intermediate metabolite in the methionine quinone pathway through multiple rounds of screening W322I\R323F The ability of the recombinant bacillus subtilis to produce heptamethylnaphthoquinone is improved, and the yield is 1.22 times that of the control strain, and the final yield reaches 186.5 mg / L. The present application not only provides a new direction for the modification of the methionine quinone pathway of bacillus subtilis, but also will be beneficial to the industrial large-scale production of heptamethylnaphthoquinone.