263 results about "Pyrroloquinoline quinone" patented technology

The present invention relates to improved variants of soluble pyrroloquinoline quinone (PQQ)-dependent glucose dehydrogenases (s-GDH), to genes encoding mutated s-GDH, to mutant proteins of s-GDH with improved substrate specificity for glucose, and to different applications of these s-GDH variants, particularly for determining concentrations of sugar, especially of glucose in a sample.

The present invention provides a mitochondrial activator containing pyrroloquinoline quinone, preferably pyrroloquinoline quinone and coenzyme Q (particularly reduced coenzyme Q) as active ingredients. In addition, the present invention provides a mitochondrial activator containing citric acid, preferably citric acid and coenzyme Q (particularly reduced coenzyme Q), as active ingredients. Since pyrroloquinoline quinone or citric acid exhibits a synergistic mitochondria activating effect with coenzyme Q (particularly reduced coenzyme Q), a high effect can be obtained even at a low dose. According to the present invention, therefore, a safer and practical mitochondrial activator is provided. The present invention further provides an agent for the prophylaxis or treatment of a disease caused by mitochondrial dysfunction.

PQQGDH having an improved substrate specificity or having an improved specific activity in an assay system using ferricyanide ion as a mediator is provided. Modified PQQGDH having the enhanced substrate specificity by introducing an amino acid mutation in a particular region of PQQGDH, and a method of enhancing the specific activity compared with a wild type in the assay system using the ferricyanide ion as the mediator by deleting, substituting, or adding one or more amino acids in an amino acid sequence of the wild type pyrroloquinoline quinone dependent glucose dehydrogenase.

The invention discloses a pyrroloquinoline-quinone-producing Hyphomicrobium strain and application thereof. The invention discloses a Hyphomicrobium denitrificans strain of which the collection number is CGMCC No.10620. The culture solution of the pyrroloquinoline-quinone-producing Hyphomicrobium FJNU-R8 has higher pyrroloquinoline quinone yield. After the strain is cultured in a culture medium using methanol as a carbon source for 4-6 days, the pyrroloquinoline quinone yield is 150-350 mg / L. The protein content is low, and the fermenting property is stable. The Hyphomicrobium disclosed by the invention can be used in producing pyrroloquinoline quinone by microbial fermentation.

The invention discloses a Hyphomicrobium sp. strain and a preparation method for pyrroloquinoline quinone. The Hyphomicrobium sp. strain 1112-NTG-2318 capable of realizing high yield of pyrroloquinoline quinine is preserved in China General Microbiological Culture Collection Center with an accession number of CGMCC No. 10709. The preparation method for pyrroloquinoline quinine comprises the following steps: fermenting the Hyphomicrobium sp. strain CGMCC No. 10709 in a fermentation medium and acquiring pyrroloquinoline quinine from fermentation broth. The fermentation titer of the preparation method in a tank with a volume of 80 m<3> reaches 1783 mg / L, so the preparation method is favorable for industrial production of pyrroloquinoline quinine.

The invention discloses a method for producing pyrroloquinoline quinone by using gluconobacter oxydans. The method comprises the following steps: (1) fermenting the gluconobacter oxydans DSM2003 to obtain fermentation liquor; (2) carrying out high-speed centrifugation on the fermentation liquor, taking supernate and enriching by using an HP-20 type macroporous resin chromatographic column to obtain an eluted fraction which is rich in pyrroloquinoline quinone; and (3) concentrating the eluted fraction which is rich in pyrroloquinoline quinone under reduced pressure and purifying by using polyamide column chromatography to obtain a pyrroloquinoline quinone pure product. According to the method, under a common culture medium and common culture conditions, the PQQ (pyrroloquinoline quinone) yield of the gluconobacter oxydans strain can reach 125mg / L. The production and preparation method is simple in condition, fast in process and convenient for industrial production in a large scale and has important meaning for industrialization of PQQ.

The invention discloses methylotrophic bacteria and a method for producing pyrroloquinoline quinine (PQQ) by fermentation thereof, and belongs to the technical field of organisms. A new bacterial strain Methylopilasp.YHT-1 is screened and separated from soil; pyrroloquinoline quinine can be produced by the bacterial strain; the bacterial strain is preserved at the China center for type culture collection on January 12, 2014; the preservation number is CCTCC NO: M2014016. The bacterial strain is subjected to aerobic culture for 3-4 days in a culture medium of taking methyl alcohol as a carbon source, so as to produce 50-113mg / L of pyrroloquinoline quinine. The condition that Methylopilasp can excessively generate PQQ and can secrete out of the cell is reported for the first time, and a new strain is provided for preparation of PQQ by a fermentation method.

The invention includes compositions comprising substantially purified pyrroloquinoline quinone, that are useful in methods for the treatment and prevention of cardiac injury caused by hypoxia or ischemia. The invention also includes methods for the treatment and prevention of cardiac injury comprising contacting a composition of the invention with a human patient.

The invention discloses oriented domestication and breeding of methylotrophic bacterium capable of producing pyrroloquinoline quinone at high yield. The invention provides an oriented domestication and breeding method for preparing a methylotrophic bacterium mutant strain capable of producing pyrroloquinoline quinone at high yield. The method comprises the following steps: 1) carrying out repeated induced culture on an initial methylotrophic bacterium used as a treatment object in a methanol-containing liquid culture medium to obtain the domesticated bacterium, wherein all the domesticated bacteria constitute a domesticated strain library; 2) fermenting the domesticated bacteria in the domesticated strain library, and collecting all the domesticated bacterium fermentation liquids; and 3) detecting pyrroloquinoline quinone in all the domesticated bacterium fermentation liquids, and selecting the single strain, the fermentation liquid of which has higher pyrroloquinoline quinone yield than the initial methylotrophic bacterium fermentation liquid, thereby obtaining the methylotrophic bacterium mutant strain capable of producing pyrroloquinoline quinone at high yield. The methanol concentration in the culture medium is gradually enhanced to perform stress domestication on the methylotrophic bacterium, thereby enhancing the breeding direct mutation rate and the pyrroloquinoline quinone yield increase amplitude.

An L-amino acid is produced by culturing a bacterium belonging to the family Enterobacteriaceae, which has an L-amino acid-producing ability and inherently has a native activity of a glucose dehydrogenase that uses pyrroloquinoline quinone as a coenzyme, but has been modified so that the activity of the glucose dehydrogenase is reduced, in a medium, and collecting the L-amino acid from the medium.

The invention discloses a combination process of nicotinamide riboside and its derivative with pyrroloquinoline quinone and its sodium salt. The combination process includes: combining any two or three of or all of NR (nicotinamide riboside) (a chloride, or nicotinamide riboside), NMN (beta-nicotinamide mononucleotide), NADH (nicotinamide adenine dinucleotide) and PQQ (pyrroloquinoline quinone disodium salt) into a formulation for making foods, health foods, cosmetics or drugs; the formulation is injected into the human body in external application, oral application or injection manner to takeeffect. The nicotinamide riboside and its derivative function to defy age; the pyrroloquinoline quinone and its sodium salt have the like function and are functionally complementary to nicotinamide riboside; the nicotinamide riboside or its derivative is combined with pyrroloquinoline quinone or its sodium salt into a formulation for making products, such as food additives, health foods, cosmetics and drugs, thereby delaying human senility, maintaining beauty or treating diseases.

The invention discloses a new strain MP688 of Methylovorus sp., of which the preservation number is CGMCC No.4096, the preservation date is August 20th, 2010, and the preservation unit is China General Microbiological Culture Collection Center. The strain MP688 is separated from soil by using a GDH (glutamate dehydrogenase) recombinase method of PQQ (pyrroloquinoline quinone). By using the Methylovorus sp. strain disclosed by the invention, the yield of PQQ can reach 125 mg / L in a conventional culture medium under conventional culture conditions; and the Methylovorus sp. strain can be used for industrial production of PQQ, and has important meaning for promoting the industrialization of PQQ.

Disclosed are: a method for easily producing pyrroloquinoline quinone in the free form without using an organic solvent or an ion-exchange resin; and high-purity crystals of the pyrroloquinoline quinone in the free form. Specifically disclosed are: a method for producing pyrroloquinoline quinone in the free form, which comprises a process wherein a solution obtained by dissolving an alkali metal salt of pyrroloquinoline quinone and having a pH of 1.5 or less is prepared so as to obtain a precipitate; and high-purity crystals of the pyrroloquinoline quinone in the free form.

It is an object of the present invention to provide a safe and highly effective preventing or ameliorating agent for skin psoriasis.The present invention provides a preventing or ameliorating agent for skin psoriasis, containing, as an active ingredient, a compound represented by general formula (I) or a salt thereof:(wherein R1, R2, and R3 simultaneously or separately represent a lower alkyl, lower alkenyl, lower alkynyl, aralkyl, araryl, or phenyl group, or a hydrogen atom).

The invention discloses a synthetic method of pyrroloquinoline quinone. The synthetic method comprises the following steps: carrying out alkali treatment on 2-methoxy-5-nitroaniline hydrochloride as a raw material, so as to obtain a compound 1; carrying out formylation on the compound 1 under a catalysis condition of an ionic liquid, so as to obtain a compound 2; adopting sodium borohydride to reduce the compound 2 to obtain a compound 3; carrying out diazotization on the compound 3, and then enabling action between the diazotized compound 3 and HBF4 to obtain a compound 4; enabling reaction of the compound 4 and 2-methylethyl acetoacetate to obtain a compound 5; treating the compound 5 with formic acid to obtain a compound 6; carrying out amid catalysis and exchange with the ionic liquid on the compound 6 to obtain a compound 7; enabling reaction of the compound 7 and 2-oxodimethyl glutaconate to obtain a compound 8; feeding hydrogen chloride to the compound 8 under the action of Cu(OAc)2*2H2O to obtain a compound 9; carrying out basic hydrolysis on the compound 9 to obtain a compound 10. The synthetic method disclosed by the invention is cheap and accessible in raw materials, stable, high in reaction yield, quick in reaction, and easy for product separation, and is environment-friendly as the catalyst can be recycled.

The invention discloses a separation and purification method of pyrroloquinoline quinone in methylotrophic bacteria fermentation liquor and application of the method. The method comprises the following steps that firstly, the fermentation liquor is enriched through macroporous resin and then sequentially passes through a buffer solution and water for elution, eluate is collected, and enrichment liquid rich in pyrroloquinoline quinone is obtained; secondly, the enrichment liquid rich in pyrroloquinoline quinone is purified through hydrophilic silica gel filler, and pyrroloquinoline quinone mother liquid is obtained; thirdly, pyrroloquinoline quinone in the pyrroloquinoline quinone mother liquid is crystallized and separated out, and crude pyrroloquinoline quinone crystals are obtained; fourthly, the crude pyrroloquinoline quinone crystals are subjected to recrystallization by using the alkali-solution and acid-isolation method, and pyrroloquinoline quinone is obtained. The experiments prove that the method has the advantages of being simple in technological operation, high in connection degree and high in product recovery rate and purity, and the method can be used for large scale industrialized production preparation of pyrroloquinoline quinone.

The invention discloses a method for synthesizing pyrroloquinoline quinone through lactobacillus fermentation. The method comprises the following steps: activating the lactobacillus plantarum PQQ-1 which is stored in an agar subculture medium through an activation culture medium, and then carrying out stationary culture of the lactobacillus plantarum PQQ-1 at a temperature ranging from 20 to 40 DEG C for 16-30h to from primary seeds; further carrying out enlarging stationary culture of the primary seeds at a temperature ranging from 20 to 40 DEG C through a primary seed culture medium for 16-30h to form secondary seeds; and further carrying out enlarging stationary culture of the secondary seeds at a temperature ranging from 20 to 40 DEG C through a secondary seed culture medium for 42-54h and then starting the fermentation stage, and carrying out anaerobic culture in a production fermentation culture medium at a temperature ranging from 20 to 40 DEG C for 54-66h to obtain the PQQ. The used lactobacillus is a food-grade safe microorganism; and the PQQ is obtained through fermentation, and is high in yield and bright in application prospect.

The invention includes compositions comprising substantially purified pyrroloquinoline quinone, that are useful in methods for the treatment and prevention of cardiac injury caused by hypoxia or ischemia. The invention also includes methods for the treatment and prevention of cardiac injury comprising contacting a composition of the invention with a human patient.

The invention refers to a medicine combination which can cure and prevent the fatty liver in mammal, especially in human and endotherm. It contains a safe and effective amount of pyrrole & quinoline quinone and / or its acceptable salt and / or ester in pharmacy as the active component, and the acceptable carrier in pharmacy. It has a remarkably curative effect on the fatty liver,

The invention discloses a method for producing pyrroloquinoline quinone table vinegar through co-culture and fermentation of microorganisms, and belongs to the technical field of table vinegar brewing. A co-culture system is constructed by saccharomyces cerevisiae, hyphomicrobium and acetobacter pasteurianus, and the table vinegar rich in pyrroloquinoline quinone is produced. The table vinegar rich in pyrroloquinoline quinone is prepared by adopting a method of co-culturing saccharomyces cerevisiae, hyphomicrobium and acetobacter pasteurianus microorganisms through the stages of alcoholic fermentation, acetic fermentation, fermentation stopping, product filtering and packaging and the like, and the concentration of pyrroloquinoline quinone is 0.05-0.2 mg / L. The method is novel, the microorganism co-culture method is adopted, nutritional factors are combined with the table vinegar, the nutritional and healthy value of the table vinegar is increased, and feasibility is achieved.

An L-amino acid is produced by culturing a bacterium belonging to the family Enterobacteriaceae, which has an L-amino acid-producing ability and inherently has a native activity of a glucose dehydrogenase that uses pyrroloquinoline quinone as a coenzyme, but has been modified so that the activity of the glucose dehydrogenase is reduced, in a medium, and collecting the L-amino acid from the medium.

A modified glucose dehydrogenase is disclosed which comprises a water-soluble glucose dehydrogenase having a pyrroloquinoline quinone as a coenzyme. At least one amino acid residue present on the surface of the water-soluble glucose dehydrogenase is replaced with arginine. The side chain of the amino acid residue is exposed at the surface of the molecule and is not expected to substantially interact with other residues. The amino acid residue is present in a region that is probably not an enzyme active site or a substrate binding site. Preferably the amino acid residue is selected from the group consisting of glutamine, asparagine, and threonine. This modified enzyme can be prepared by recombinant processes and recovered efficiently.

The present invention relates to a method for lowering activity with respect to maltose in glucose measurement comprising a step of reacting modified pyrroloquinoline quinone dependent glucose dehydrogenase subjected to amino acid sequence modification, wherein pyrroloquinoline quinone dependent glucose dehydrogenase is reacted in the presence of at least one type of substance selected from the group comprising succinic acid, malonic acid, glutaric acid, malic acid, phthalic acid, 2-ketoglutaric acid, 3,3-dimethylglutaric acid, pimeric acid, suberic acid, adipic acid, maleic acid, potassium chloride, ammonium chloride, diammonium hydrogen citrate, L-lysine, taurine, calcium glycerate, amino-n-butyric acid, sodium glycolate, sodium alpha-ketoglutarate, fumaric acid, glycine, glutamic acid, serine and citric acid.

The invention discloses application of pyrroloquinoline quinone (PQQ) in preparing products for preventing and / or treating male reproductive dysfunction or improving male reproductive function. Research results of the invention show that the pyrroloquinoline quinone can obviously reduce Bmi1- / -mouse spermatogenic cell DNA damage and finally can improve Bmi1- / -mouse spermatogenic cell proliferation and inhibit spermatogenic cell apoptosis. According to the novel application disclosed by the invention, the pyrroloquinoline quinone is applied to treat male sterility for the first time, a more ideal treating effect is obtained, a novel preventing and treating choice is provided for treating male sterility in clinical application, and very high application valve is achieved.

The invention provides an application of a pyrroloquinoline quinone (PQQ) active substance in preparation of medicaments for treating and / or improving a diabetic complication, namely diabetic foot and further provides a health-care product composition or a pharmaceutical composition containing pyrroloquinoline quinone. The active substance, namely the pyrroloquinoline quinone in the composition provided by the invention has significant treatment and / improvement action against the diabetic complication, namely the diabetic foot by improving nerve conduction velocity, reducing sorbitol content, increasing the open number of capillaries, reducing whole blood viscosity, enhancing blood flow, increasing neural blood flow, promoting blood circulation, improving microcirculation of blood vessels and reducing gangrene ulcers of the foot. Therefore, the pyrroloquinoline quinone has great application prospects in prevention, treatment and / or improvement of occurrence and development of the diabetic complication, namely the diabetic foot.

The invention discloses a method for increasing the pyrroloquinoline quinone yield, belonging to the field of biotechnology. In the invention, according to analysis of trophic factors light green yellow pseudomonas produced pyrroloquinoline quinone, the yield of pyrroloquinoline quinone can be effectively increased by controlling the content of specific amino acid and vitamin content in a medium. By adopting the method disclosed by the invention, the pyrroloquinoline quinone yield of a strain can be increased by 15-89%. Moreover, the method disclosed by the invention is simple and easy to control and applicable to industrial application.

The present disclosure relates to compounds of Formula (I) and process of obtaining the same. Said compounds of Formula (I) is employed in the syntheses of pyrroloquinoline quinone (PQQ), 4,5-dioxo-4,5-dihydro-1H-pyrrolo[2,3-f]quinoline-2,7,9-tricarboxylic acid 2-allyl ester, 5-ethoxy-5-hydroxy-4-oxo-4,5-dihydro-1H-pyrrolo[2,3-f]quinoline-2,7,9-tricarboxylic acid, 5-hydroxy-6-(1,1,4-trioxo-1 lambda*6*-1,2,5-thiadiazolidin-2-yl)-1H-indole-2-carboxylic acid and its pharmaceutically acceptable salts. Said syntheses of compounds via compounds of Formula (I) as intermediates employ minimum number of steps resulting in a shorter process and has improved efficiency along with many other advantages.