197 results about "Cyclase" patented technology

The invention relates to the field of biological engineering and technology, in particular to an enzyme for synthesizing glabridin, a biological synthesis method of glabridin and a bacterial strain. The invention discloses a series of glabridin synthesizing enzymes (phenylalanine ammonialyase, cinnamic acid-4-hydrogenase, tyrosine ammonialyase, p-coumaric acid coenzyme A ligase, chalcone synthase, chalcone reductase, chalcone isomerase, isoflavone synthase, 2-hydroxyisoflavone dehydratase, 4 '-oxymethyltransferase, 4'-hydroxymethyltransferase and 4 '-hydroxymethyltransferase) derived from glycyrrhiza glabra for the first time. ) can be used in the preparation of a compound (e.g., isoflavone 2 '-hydrogenase, isoflavone reductase, Vissisoketoreductase, isoprenyltransferase, pterocarpin reductase, oxidative cyclase, demethylase, cytochrome P450 reductase, and the like). The de novo synthesis of the glabridin in a microbial cell factory is realized for the first time, and the application prospect of the glabridin biological manufacturing industry is greatly promoted.

The invention discloses a high-yield delta-tocotrienol genetically engineered bacterium and application thereof, and belongs to the technical field of gene recombination. The invention discloses a genetically engineered bacterium for high yield of delta-tocotrienols, which is characterized in that yarrowia lipolytica ATCC MYA-2613 is used as an original strain, and a ku70 gene is knocked out; the gene is used for expressing and coding 4-hydroxyphenylpyruvate dioxygenase, the gene is used for coding uromelanic acid phytate transferase and the gene is used for coding tocopherol cyclase; intensifying a shikimic acid pathway; a mevalonic acid pathway is strengthened; an isopentenol path is introduced; carrying out protein fusion on the HPD and the HPT; truncating a signal peptide of a gene encoding a tocopherol cyclase and back-filling a leucine defect. The genetically engineered bacterium constructed by the invention has industrial stability, can be amplified by more than 10 times in a 2L fermentation tank level compared with a shake flask level, can produce 616.45 mg / L of delta-tocotrienol after being cultured for 240 hours, and has industrial application prospects.

The present invention relates to the use of stimulators of soluble guanylate cyclase (sGC), pharmaceutically acceptable salts thereof and pharmaceutical formulations or dosage forms comprising them, alone or in combination with one or more additional agents, for the treatment of various mitochondrial diseases wherein sGC stimulation is increased, or the concentration of nitric oxide (NO) or cyclic 3 ', 5'-guanosine monophosphate (cGMP), or both, is increased, or both, or both. Or an up-regulation of the NO-sGC-cGMP pathway is desirable. Compounds useful in the methods of the invention are compounds of Formula I or pharmaceutically acceptable salts thereof.

The invention discloses a Yarrowia lipolytica engineering bacterium for producing astaxanthin in a peroxisome compartment and application of the Yarrowia lipolytica engineering bacterium. A complete astaxanthin synthesis path is expressed in host bacteria by recombinant Yarrowia lipolytica; comprising geranyl geranyl diphosphate synthase CrtE, phytoene synthetase / lycopene cyclase CrtYB, phytoene desaturase CrtI, 3-hydroxy-3-methylglutaryl CoA reductase tHMGR, beta-carotene ketolase CrtW and beta-carotene hydroxylase CrtZ, and the enzymes are relocated into peroxisome, so that the enzyme activity of the peroxisome is improved, and the enzyme activity of the peroxisome is improved. The number of expression hemoglobin genes VHb and peroxisome is increased, the surface area of the hemoglobin genes VHb is enlarged, and one or more of enzymes PEX11, PEX19, DNM1 or PHO85 for prolonging the service life of the hemoglobin genes VHb and peroxisome is / are obtained; the astaxanthin production performance of the recombinant strain is verified based on cytoplasm engineering and subcellular organelle compartment engineering, and the astaxanthin production capacity of the yarrowia lipolytica is further improved.

The invention discloses a nitrogen-containing heterocyclic compound and application thereof. The nitrogen-containing heterocyclic compound has a structure as shown in a formula (I). The compound provided by the invention has an excellent glutamine cyclase inhibition effect, can be applied to treatment of tumors, immune diseases, neurological diseases or aging, and has a good application prospect.

The invention discloses a culture medium of a lacrimal gland organ and a culture method of the lacrimal gland organ, and the culture medium of the lacrimal gland organ is composed of a basic culture medium, R-spodin3, Noggin, B27, N-acetyl-L-cysteine, trehalose, vitamin C, glutathione, an adenylate cyclase activator, A8301, PGE2 and FGF10. The culture medium for lacrimal gland organ culture, provided by the invention, is prepared from various cell factor components according to an optimized proportion, and is simple and reasonable in components and rich in nutrition; when the culture medium is used for culturing the lacrimal gland organoid, the formation and growth of the organoid can be promoted, an in-vivo complex microenvironment is further simulated, the organoid is closer to an in-vivo physiological state, an organoid model is better constructed so as to evaluate the influence of drugs on lacrimal gland functions, and a new platform is provided for drug screening; and a new way is opened up for follow-up treatment of diseases such as xerophthalmia in the future.

The invention discloses a method for detecting the protein catalytic function of carrot lycopene beta-cyclase, and belongs to the technical field of protein catalytic function detection. According to the method for detecting the catalytic function of the carrot lycopene beta-cyclase protein, escherichia coli is used as a heterologous expression host, a lycopene substrate is accumulated in the escherichia coli by expressing pACCRT-EIB plasmids, then lycopene epsilon-cyclase protein and lycopene beta-cyclase are constructed on the same carrier for co-expression, and the lycopene beta-cyclase protein is obtained. A monocyclic carotene intermediate product in the process of converting lycopene into carotene is introduced into a reaction system so as to detect the substrate preference and functional difference of two lycopene beta-cyclase isoenzymes. The method is easy to operate, expression in an escherichia coli system is convenient and rapid, various lycopene beta-cyclase catalytic substrates except lycopene can be provided, and the catalytic function of lycopene beta-cyclase protein can be identified more accurately.

The invention discloses a saccharomyces cerevisiae strain for de novo synthesis of fucoxanthine precursor neoxanthine and application of the saccharomyces cerevisiae strain, and belongs to the technical field of biology. According to the recombinant saccharomyces cerevisiae disclosed by the invention, the 3-hydroxy-3-methylglutaryl coenzyme A reductase, the isopentenyl pyrophosphate isomerase, the farnesyl pyrophosphate synthase and the endoplasmic reticulum size regulation factor are expressed in an enhanced manner; according to the method, geranyl geranyl diphosphate synthase, phytoene dehydrogenase, 15-cis-phytoene synthase, bifunctional lycopene cyclase / phytoene synthase, beta-carotene hydroxylase, zeaxanthin epoxidase truncated mutant and violaxanthin deep oxidase-like protein are subjected to heterologous expression, so that the zeaxanthin deep oxidase-like protein is obtained; the ROX1 and GAL80 genes in the saccharomyces cerevisiae are knocked out, and the expression of squalene synthase ERG9 is down-regulated. When the strain is subjected to shake flask fermentation, the contents of neoxanthine and violaxanthin of the strain respectively reach 1.78 mg / L and 26.77 mg / L, so that the strain has a wide application prospect.

The invention provides a compound serving as a glutamine cyclase inhibitor and application of the compound, and belongs to the field of medical chemistry. The structure of the compound is shown as a formula I. The compounds have good inhibitory activity on sQC and gQC enzymes and mutants thereof, and the half inhibitory concentration of most compounds reaches the sodium mole level, so that the compounds can be used for preparing glutamine cyclase inhibitors. In addition, the compounds can also be used for preparing medicines for preventing and treating diseases related to glutamine cyclase. The invention lays a material basis for research and development of glutamine cyclase inhibitor drugs, and has a good application prospect. Formula I.

The present disclosure relates to stimulators of soluble guanylate cyclase (sGC), pharmaceutically acceptable salts thereof and pharmaceutical formulations comprising them and their use, alone or in combination with one or more additional agents, for the treatment of various diseases wherein the concentration of nitric oxide (NO) and / or the concentration of cyclic guanosine monophosphate (cGMP), or both, is increased, such as, for example, the concentration of nitric oxide (NO) and / or the concentration of cyclic guanosine monophosphate (cGMP). Or regulation of the NO pathway is desirable. In some embodiments, the compound is a compound of Formula I or a pharmaceutically acceptable salt thereof. # imgabs0 #

The invention belongs to the technical field of enzyme mutants, and discloses a mutant of lycopene cyclase and application of the mutant, glutamic acid at the 321 site of wild type lycopene cyclase which is derived from arabidopsis and has an amino acid sequence as shown in SEQ ID NO.1 is mutated into lysine, phenylalanine at the 319 site of the wild type lycopene cyclase is mutated into leucine, cysteine at the 323 site of the wild type lycopene cyclase is mutated into alanine, and the mutant of the lycopene cyclase is obtained. 1, and the lycopene cyclase mutant with the amino acid sequence as shown in SEQ ID NO. 2 is obtained. BTS1, CrtB and CrtI are integrated into a saccharomyces cerevisiae BY4741 strain, and a chassis strain ZA1 for stably producing alpha-carotene precursor lycopene is constructed; mutant expression plasmids are constructed, positive clone strains are screened out, and the epsilon-cyclization ability of mutant enzymes is remarkably higher than the beta-cyclization ability; the method comprises the following steps: by taking lycopene as a substrate, constructing an expression vector lipid droplet surrounding protein PET10; gene PAH1, DGA1 and Cat2 related to TAG synthesis and perilipid droplet protein PET10 are constructed on an expression vector and converted into ZA1, and when lycopene is used as a substrate, the efficiency of catalytic production of alpha-carotene is improved.

This invention relates to the application of exosomes in reversing tumor resistance to immune checkpoint (PD-1) inhibitors. A novel application of exosomes in the preparation of pharmaceutical compositions reversing tumor resistance to immune checkpoint inhibitors is disclosed. Exosomes generated from tumor cells recombinantly expressing adenylate cyclase 7 (ADCY7) can significantly reverse tumor resistance to PD-1 inhibitors, and the exosomes exhibit a synergistic effect with the PD-1 inhibitors.

The invention discloses an application of polygonatum cyrtonema cyclase PcAS1 in promoting synthesis of hematoxylin A. The cyclase PcAS1 is derived from polygonatum cyrtonema, a high isoflavone compound hematoxylin A is synthesized in a plant body, and according to the application, a recombinant plant expression vector for expressing a PcAS1 gene is constructed and transformed into the plant to realize synthesis of the hematoxylin A. Based on multiple omics data such as whole genome, re-sequencing and transcriptome of polygonatum cyrtonema, key cyclase PcAS1 closely related to homoisoflavone biosynthesis is systematically mined. Through a tobacco transient expression system and a polygonatum cyrtonema hairy root transformation system, it is proved for the first time that cyclase PcAS1 can significantly promote synthesis of high isoflavone hematoxylon A, and biosynthesis of hematoxylon A is achieved in polygonatum cyrtonema for the first time.

The invention discloses a biosynthesis method and application of cannabinoid compounds. The specific method comprises the following steps: constructing a non-reductive polyketide synthase (NRPKS) AtnG from a fungus Arthrinum sp.NF2194, an isopentenyl transferase NphBV49W / Y288P from an actinomycetes Streptomyces sp.CL190, a cyclase Svz9 from an actinomycetes Streptoviensis NA431, a GPP synthase AgGPPS from a plant Abis grandis and a terpene synthesis enhancement path MEV into a heterologous expression vector, and sequentially introducing the heterologous expression vector into escherichia coli, so as to successfully realize the production of the cannabinoid compounds OA, CBGA and CBCA. The cannabinoid compound disclosed by the invention has important medical and industrial values.

The invention relates to a crystal form of a compound shown as a formula I or a pharmaceutically acceptable salt thereof and a preparation method of the crystal form of the compound shown as the formula I or the pharmaceutically acceptable salt thereof. The compound of the formula I or the pharmaceutically acceptable salt thereof is used for treating cardiovascular diseases, endothelial dysfunction, diastolic dysfunction, atherosclerosis, hypertension, heart failure, pulmonary arterial hypertension (WHO levels I, II, III, IV), angina pectoris, thrombus, restenosis, myocardial infarction, stroke, cardiac insufficiency, fibrosis, pulmonary hyperplasia, erectile dysfunction, asthma, chronic kidney disease, diabetes, and the like. The composition can be used for treating liver cirrhosis, chronic obstructive pulmonary disease (COPD), acute respiratory distress syndrome, acute lung injury, pulmonary fibrosis, cystic fibrosis or interstitial lung disease.

The invention belongs to the technical field of biological medicines, and particularly relates to a multifunctional peptide and application thereof in anti-inflammatory, antibacterial, mucous membrane repair and intestinal secretion promotion. Specifically, the invention designs a polypeptide and an analogue which have guanylate cyclase C receptor (GUCY2C) agonist activity and anti-inflammatory, antibacterial and mucous membrane repair functions at the same time. The polypeptide can be combined with GUCY2C, stimulate generation of cyclic guanosine monophosphate (cGMP) in cells, promote intestinal fluid secretion, inhibit release of proinflammatory factors, relieve inflammatory response, promote repair of intestinal injury mucosa, inhibit growth of opportunistic pathogenic bacteria and accelerate intestinal homeostasis recovery after intestinal preparation. The composition can effectively promote intestinal tract cleaning, does not cause intestinal inflammation and mucosal lesion, has no hemolytic toxicity and cytotoxicity, is high in safety, and has a good clinical application prospect.

This invention discloses a genetically engineered bacterium capable of synthesizing large quantities of natural cyclic diguanylate (c-di-GMP) and its applications. It involves knocking out the major phosphodiesterase gene chr1_233 as described in claim 1 within the sphingobium xenophagum C2 strain. This invention rationally designs and modifies the c-di-GMP synthesis and degradation pathways and the inhibitory site of major diguanylate cyclase activity in heterologous sphingobium-consuming bacteria, constructing a series of genetically engineered bacteria capable of synthesizing large quantities of natural c-di-GMP. This provides the industry with a green, simple, economical, and efficient method for producing natural c-di-GMP, and offers a series of high-quality and inexpensive raw materials for the preparation of STING agonists or immune adjuvants, or for the preparation of drugs for treating diseases related to STING protein function.

Compounds (I) are provided, where R1 and R2 are H or (C1-C3)-alkyl; X is a linear methylene chain of formula —[CH2]n— with n=0, 1 or 2, or a biradical from a branched saturated (C2-C4)-alkylene chain; and A is either a C-radical from a non-aromatic polycyclic 6- to 15-membered carbocyclic ring system, or a C-radical from a polycyclic 6- to 15-membered heterocyclic ring system having one or two O, S or N; wherein the C-radicals are unsubstituted or substituted. Compounds (I) are simultaneously inhibitors of soluble epoxide hydrolase and inhibitors of glutaminyl cyclase. Besides, they reduce the levels of pro-inflammatory cytokines in LPS stimulated BV2 cells, display low cytotoxicity, and have good BBB permeability. Thus, they are useful as multitarget compounds for the prevention or treatment of Alzheimer's disease.

The present invention relates to compounds of formula (I) or formula (II) or pharmaceutically acceptable salt, solvate or hydrate thereof, wherein said compound of formula (I) and said compound of formula II each comprises at least one ONO2 or ONO moiety; R1 is C1-C3alkyl; R2 is H, C1-C6alkyl, C3-C6cycloalkyl, C1-C2alkoxy, C2-C4alkenyl; R3 is C1-C4alkyl optionally substituted with C1-C2alkoxy, C3-C4cycloalkyl, C2-C4alkenyl; R4 and R5 are each independently H or C1-C6alkyl optionally substituted with F, OH, ONO, ONO2, COOH, C1-C3alkoxy, C3-C6cycloalkyl; or together with the nitrogen atom to which they are attached form a heterocyclic ring, wherein preferably said heterocyclic ring is selected from aziridine, azetidine, pyrollidine, piperidine, morpholine, piperazine, homo-piperazine, 2,5-diazabicyclo[2,2,1]heptane and 3,7-diazabicyclo[3,3,0]octane, wherein said heterocyclic ring is optionally substituted with independently one or more R6; R6 is C1-C6alkyl optionally substituted with independently one or more halogen, OH, ONO, ONO2, C1-C3alkoxy, C1-C3haloalkoxy, COOR7, NR8R9, C═NR10; R7 is H, or C1-C4alkyl optionally substituted with F, OH, ONO, ONO2, NR8R9; R8 and R9 are independently H, or C1-C4alkyl optionally substituted with ONO, ONO2; R10 is C1-C4alkyl optionally substituted with F, ONO, ONO2; C3-C6cycloalkyl; pharmaceutical compositions thereof, and their use in methods of treating or preventing a disease alleviated by inhibition of PDE5 in a human or in a non-human mammal.

The present disclosure provides combinations of GLP-1 receptor agonists, and guanylate-cyclase C (GC-C) receptor agonists, and pharmaceutical compositions comprising the same. Also described are methods of making and using the combination, which can be used to treat or prevent of one or more side effects of a GLP-1 receptor agonist.

A pharmaceutical composition including a soluble guanylate cyclase stimulator or salt thereof for use in the treatment of a medical condition comprising a chronic vascular dysfunction occurring or persisting after onset of an infection in a human subject. In preferred embodiments, the pharmaceutical composition comprises a soluble guanylate cyclase stimulator or salt thereof, preferably vericiguat, for use in the treatment of a chronic vascular dysfunction associated with vascular constriction, endothelial dysfunction and / or hypo-perfusion.

The invention discloses a saccharomyces cerevisiae genetically engineered bacterium for efficiently synthesizing mogroside V based on multi-compartment multi-metabolic pathways and a construction method of the saccharomyces cerevisiae genetically engineered bacterium. The preparation method comprises the following steps: anchoring an MVA pathway and a squalene synthesis pathway on the surface of a peroxisome membrane, and enabling mogrol synthetase and epoxy squalene cyclization enzyme ERG1 to form a linear polywheat complex through polypeptide interaction; then, through overexpression of MVA pathway key enzyme and introduction of an isopentenol utilization pathway, supply of precursor isopentenyl pyrophosphate and dimethyl allyl pyrophosphate in cytoplasm is increased, and squalene epoxidase ERG1 and mogrol synthetase are anchored in transmembrane ABC efflux protein PDR11 through polypeptide interaction; then anchoring the mogrol synthetase in the lipid droplet subcells; finally, the glucosyltransferase is displayed on the surface of the yeast cell wall through an alpha-lectin system, so that extracellular mogrol is converted into MG-V. The engineering bacterium improves the yield of mogroside V synthesized from the beginning in microorganisms, and has great industrial application value.

This invention discloses a guanylate cyclase-C (GC-C) activating peptide and its applications, belonging to the field of bioactive peptide technology. The amino acid sequence of the GC-C activating peptide is PFPGPIPN. The GC-C activating peptide of this invention can significantly activate GC-C activity, increasing intracellular cGMP levels. Furthermore, the GC-C activating peptide of this invention can alleviate constipation and abdominal pain symptoms in mice, thereby effectively preventing or improving irritable bowel syndrome (IBS). Therefore, the GC-C activating peptide provided by this invention has advantages such as high activity, safety and non-toxicity, and good water solubility. It can be used as a candidate molecule for alternative or adjuvant drugs in the clinical treatment of IBS, and also as a health product or food to improve constipation or abdominal pain.

Methods and materials for treating mammals having cardiovascular and / or metabolic disease are provided herein. For example, methods and materials for treating mammals having cardiovascular and / or metabolic disease by administering (a) an analog of a natriuretic peptide (NP), such as atrial natriuretic peptide (ANP) or dendroaspis natriuretic peptide (DNP), and (b) a glucagon-like peptide-1 (GLP-1) receptor / glucose-dependent insulinotropic polypeptide (GIP) receptor agonist are provided herein. The NP analogs used in the methods provided herein can be less than 20 amino acids in length and, in some cases, can have one or more variations in their ring portion (as compared to wild type ANP or DNP) that affect the potency of the analog in activating the particulate guanylyl cyclase (GC-A) receptor.

The present invention relates to a bifunctional terpene synthase and its mutants, as well as the catalytic products. The amino acid sequence of the bifunctional terpene synthase is shown in SEQ ID NO.1. The bifunctional terpene synthase mutant is a protein composed of a new amino acid sequence formed by mutating the leucine at the 89th position of the amino acid sequence shown in SEQ ID NO.1 to glutamine or asparagine. The bifunctional terpene synthase FoFS to be modified in the present invention simultaneously has an isopentenyl transferase domain and a terpene cyclase domain, and has the dual functions of catalyzing the head-to-tail connection and cyclization of the terpene synthesis precursors DMAPP and IPP. FoFS has sesquiterpene products with multiple different ring systems. By modifying FoFS, the mutant protein can catalyze sesquiterpene compounds with other complex ring systems, which is of great significance for expanding the library of terpene natural products and adding more potential drug-value terpene compounds to the reserve force.

A pharmaceutical composition includes a combination of a guanylate cyclase C (GUCY2C) agonist and a short-chain C2 to C5 fatty acid and / or a salt and / or a prodrug thereof in a therapeutically effective amount and one or more pharmaceutically acceptable excipients as well as to a method of preventing and / or treating colorectal tumorigenesis and / or carcinogenesis and / or chronic intestinal inflammation and / or cystic fibrosis related gastrointestinal manifestations by administering to a patient, who has developed or is at risk to develop colorectal tumorigenesis and / or carcinogenesis and / or chronic intestinal inflammation and / or cystic fibrosis related gastrointestinal manifestations, a therapeutically effective amount of a combination of a GUCY2C agonist and a short-chain C2 to C5 fatty acid or a salt or a prodrug thereof.

The present invention relates to a method of treating a mitochondrial disease in a patient in need thereof by administering Compound (I), a stimulator of soluble guanylate cyclase (sGC) at certain dosages either alone or in combination therapy.

The present disclosure relates to the use of stimulators of soluble guanylate cyclase (sGC), pharmaceutically acceptable salts thereof, and pharmaceutical preparations or dosage forms containing them, alone or in combination with one or more additional agents, for the treatment of various mitochondrial diseases, in which increased sGC stimulation, or increased concentrations of nitric oxide (NO) or cyclic guanosine 3',5'-monophosphate (cGMP), or both, or upregulation of the NO-sGC-cGMP pathway is desired. Compounds useful in the methods of the present invention are of Formula I or pharmaceutically acceptable salts thereof: