62 results about "Heme" patented technology

The invention relates to the technical field of medicines, and discloses an application of combination of bufalin and heme in preparation of a cancer treatment medicine. According to the invention, by means of an active metabonomics means, a key active metabolite-heme is locked through exogenous library screening and endogenous analysis, and the drug effect of bufalin is regulated by means of the active metabolite. Through verification, it is found that heme can cooperate with the synergistic drug effect in various cancer cell lines, and a new technical normal form based on an active metabolome for improving the drug effect of bufalin is provided. The pharmaceutical composition can improve the apoptosis induction ability of bufalin to different cancer cells, and provides powerful guiding significance for developing a novel treatment scheme in clinical medicine.

The present disclosure describes anti-inflammatory properties of red blood cell extracellular vesicles, e.g., as mediated by heme, hemoglobin and / or phosphatidylserine content. The present disclosure describes technologies useful for the treatment of inflammatory diseases, disorders, or conditions (e.g., atherosclerosis).

The present invention relates to the fields of biopharmaceuticals and biochemical engineering, and particularly relates to a method for preparing chiral β-trifluoromethyl alcohol compounds using non-heme iron enzyme BsQueD or its mutants. Using non-heme iron enzyme BsQueD or its mutants as catalysts, trifluoromethyl ketone as a substrate, and phenylsilane as a hydrogen source, chiral β-trifluoromethyl alcohol is obtained after the reaction. The amino acid sequence of the non-heme iron enzyme BsQueD is shown in SEQ ID NO.1. It has a high conversion rate (70%) for the substrate trifluoromethyl ketone, high optical activity of the product β-trifluoromethyl alcohol (e.e. value greater than 90%), and high yield, greatly reducing the production cost.

The invention relates to gene modification, in particular to a method for improving the catalytic capacity of engineering P450 peroxygenase hydrogen peroxide and application of the method. The invention relates to a method for improving hydrogen peroxide catalytic capacity of P450 enzyme, which comprises the following steps of: identifying a channel of the P450 enzyme by using Caver Web 1.0 software by taking a heme cofactor of the P450 enzyme as a starting point; the method comprises the following steps: selecting a target channel with the channel length of 10-30, the curvature of 0-2 and the diameter of 0-3 as a target channel, then selecting amino acid of which the side chain faces the channel at an inlet or a bottleneck of the target channel as key amino acid, and then performing specific mutation on one or more key amino acids, further, at least one channel capable of improving the hydrogen peroxide catalytic capability of the P450 enzyme is obtained, so that the catalytic capability of the P450 enzyme is improved under the catalysis of hydrogen peroxide. The catalytic activity of a plurality of mutants obtained by the invention is compared with that of a wild enzyme, and the channel with high catalytic activity is an H2O2 channel. The application of the method can improve the hydrogen peroxide utilization capability of the P450 enzyme, and further improves the activity of the catalytic substrate of the P450 enzyme.

The present invention refers to Heme-Regulated Inhibitor (HRI) Kinase agonists or activators, or pharmaceutical composition comprising thereof, for use in a method for the treatment of a neurological disorder or for use as a neuroprotective agent.

This application discloses a recombinant bacterial strain that highly expresses nonspecific peroxygenase, its construction method, and its applications, relating to the field of microbial engineering technology. The strain of this invention uses *Escherichia coli* BL21(DE3) as the host. First, the key gene *hemA* for heme synthesis is integrated into the genome using the CRISPR-transposon system. Then, a chassis engineered strain A2 with 2 copies of optimal integration is screened and co-transformed with the rDcaUPO-A161C expression plasmid and the molecular chaperone plasmid pG-KJE8. The intracellular heme level of this strain is increased by 2.84 times compared to the original strain, and the total enzyme activity of rDcaUPO-A161C is cumulatively increased by 8.9 times and the specific enzyme activity is increased by 6.21 times compared to the wild type. This fundamentally solves the problems of insufficient cofactors, low folding efficiency, and poor activity in heterologous expression of nonspecific peroxygenases, and can be used for the efficient production of nonspecific peroxygenases.

The application discloses a HtsA-SLOdm fusion protein and application thereof in preparation of a group A streptococcus vaccine. The HtsA-SLOdm fusion protein is obtained by fusing and recombining a heme transport lipoprotein (HtsA) and a streptolysin protein containing a double-mutation site (SLOdm), can induce production of HtsA protein and SLOdm protein specific IgG antibodies in serum, has strong immunogenicity, can induce high-level IgG1 and IgG2a antibody titers, and the humoral immune response in inducing production of SLOdm protein specific IgG antibodies in serum is stronger than that of a single protein. In addition, the HtsA-SLOdm fusion protein can promote skin wound healing of mice, reduce bacterial content and inflammatory response at a skin infection site of the mice, accelerate wound healing, has almost no hemolytic activity, has high safety, and can be used for preparing a group A streptococcus vaccine.

This invention discloses a genetically engineered bacterium for synthesizing carotenoids, its construction method, and its applications, belonging to the field of genetic engineering technology. The genetically engineered bacterium uses a carotenoid-producing yeast strain as the starting material. The gene ROX1, encoding a heme-dependent hypoxia gene repressor, is knocked out, while the expression of genes STB5 (encoding a transcription factor involved in NADPH regeneration), DID2 (encoding a class E protein in the vacuole protein sorting pathway), and VOA1 (encoding an endoplasmic reticulum protein playing a role in the assembly of the V0 region of V-ATPase) is upregulated. This invention promotes carotenoid synthesis and significantly increases carotenoid yield through combined regulation of gene expression outside the carotenoid synthesis pathway, demonstrating promising application prospects.

The present disclosure relates to yeasts, in particular Yarrowia cells, with enhanced production of heme. Also provided are methods for enhanced production of heme with Yarrowia cells.

Provided are a recombinant microorganisms having a genetic modification that increases the expression of bacterioferritin, a composition comprising the recombinant microorganism for use in reducing a nitrogen oxide concentration in a sample, and a method of reducing a nitrogen oxide concentration in a sample.

This application relates to the field of biotechnology, and particularly to primer pairs, primer-probe compositions, PCR premixes, kits, and methods for detecting multiple respiratory pathogens. Nucleic acid sequences are shown in primer pairs as indicated by SEQ ID NO: 1-2, 4-5, 7-8, 10-11, 13-14, 16-17, 19-20, 22-23, 25-26, 28-29, 31-32, 34-35, 37-38, and 40-41. For target respiratory pathogens, this application designs specific amplification primer pairs, paired with suitable probes, enabling the simultaneous detection of eight targets. Furthermore, the detection process avoids interference from heme, trimethoprim, sulfamethoxazole, amphotericin B, itraconazole, fluconazole, azithromycin, adrenaline, and lidocaine hydrochloride in the test sample. It exhibits good anti-interference properties and shows no cross-reactivity with *Rhodococcus equi*, *Candida tropicalis*, and *Candida krusei*.

This invention discloses a magnetic hydrogel-encapsulated bacterial biosensor platform and its application, belonging to the fields of synthetic biology and clinical diagnostics. The hydrogel delivery and recovery system comprises a whole-cell biosensor encapsulated in hydrogel and magnetic microparticles. The hydrogel is constructed from sodium alginate and calcium chloride, and the biosensor is a heme-sensing bacterium. More specifically, this invention uses sodium alginate hydrogel to encapsulate engineered bacteria YES601 and magnetic Fe3O4 microparticles to form magnetic hydrogel microspheres. These microspheres are delivered to the intestine as biosensors and can emit light in response to heme. The intact microspheres are then efficiently recovered from feces via magnetic adsorption for detection, improving the efficiency and safety of whole-cell biosensors in detecting intestinal bleeding. The overall design optimization strategy of the hydrogel delivery and recovery system provided by this invention can improve diagnostic efficiency and practical application value.

This invention provides a non-heme chloride peroxidase and its application, belonging to the field of genetic engineering. The non-heme chloride peroxidase... Ro CPO, whose amino acid sequence is SEQ ID NO.2; the non-heme chlorperoxidase Ro The nucleotide sequence of the CPO encoding gene is SEQ ID NO.1; the non-heme chlorperoxidase... Ro Application of CPO in the preparation of 5-hydroxymethyl-2-furanic acid and / or 2,5-furanic acid.

The invention relates to the technical field of biology, and particularly discloses a construction method and application of a recombinant Escherichia coli strain with high yield of bilirubin, and the construction method of the strain comprises the following steps: knocking out a heme precursor efflux gene in Escherichia coli, co-expressing the following modules in the Escherichia coli, introducing a gene gdha for enhancing coenzyme factor circulation, and carrying out expression to obtain the recombinant Escherichia coli strain with high yield of bilirubin. According to the method disclosed by the invention, heme precursor exclusion genes are knocked out in escherichia coli, meanwhile, heme oxygenase and biliverdin reductase are modularly assembled by adopting a protein covalent coupling technology, and the yield of bilirubin is increased by increasing synthesis of coenzyme.

Disclosed are compositions and methods for activating NLRP12 and / or NLRC5 and inflammatory cell death using heme and at least one PAMP and / or DAMP molecule.Also provided are screening assays for identifying NLRP12 and / or NLRC5 inhibitors, and the use of such inhibitors in treating or alleviating NLRP12-mediated or NLRC5-mediated inflammation associated with hemolytic disease, infectious disease, cancer, or inflammatory syndrome.

The application discloses a detection kit based on a colorimetric method and application thereof, and the detection kit comprises two membrane protein recognition chains T1 and T2, two DNA hairpins H1 and H2 required for forming G-quadruplex, hematin required for a color developing reaction, hydrogen peroxide (H2O2) and ABTS. The membrane protein recognition chains T1 and T2 can anchor membrane proteins, and adjacent hybridization occurs when the membrane proteins dimerize, thereby triggering catalytic hairpin self-assembly of H1 and H2 to form a large number of G-quadruplex structures; when H2O2 exists, G-quadruplex / hematin formed by combining G-quadruplex and hematin can catalyze oxidation of ABTS to generate a color developing reaction. The detection kit has the characteristics of high sensitivity, good specificity, low cost, low dependence on equipment and simple operation in the detection of cell membrane protein dimerization, and also has good application performance in cell signal pathway visualization and rapid diagnosis of tumor cells at a bedside.

The invention discloses a high-heme-yield escherichia coli engineering bacterium as well as a construction method and application thereof, and belongs to the technical field of synthetic biology. According to the engineering bacteria disclosed by the invention, eight genes in a heme C4 synthetic route are designed into two fusion multienzyme complexes, short peptide Linker is adopted for connection, escherichia coli preference codon optimization is performed, and the enzyme synergy efficiency is remarkably improved; and meanwhile, constructing a heme efflux protein recombinant plasmid, and co-transforming the heme efflux protein recombinant plasmid and the fusion multienzyme complex plasmid into an escherichia coli host to form a double-plasmid co-expression system. The engineering bacterium takes glucose / glycerol as a substrate and lactose as an inducer for fermentation, the shake flask yield of the heme reaches up to 865 mg / L and is increased by more than one time compared with the prior art, expensive intermediates such as glutamic acid / glycine do not need to be added, and the bottleneck of intracellular accumulation inhibition is effectively broken through. The fermentation product is directly extracted through the acetic acid solvent, the process is simple, the cost is low, the method is suitable for large-scale production of safe and high-purity heme, and a green biological manufacturing solution is provided for the fields of artificial meat, medicine and health care products.

This application provides a method and kit for detecting pathogenic microorganisms based on the properties of tetrastranded DNA oxidoreductase. The method utilizes isothermal amplification technology to amplify genomic fragments containing tetrastranded DNA / RNA sequences in the pathogenic microorganism genome, and then performs a colorimetric reaction by adding heme and oxidoreductase substrates, achieving rapid and visual detection of pathogenic microorganisms.

The application belongs to the technical field of biosensors and provides an electrochemical biosensor for detecting ochratoxin A based on DNA tetrahedron, which comprises an A-P probe, a gold electrode with a modified capture probe CP, a circular template CT, phi 29 polymerase, dNTPs, Mg 2+ , hemoglobin, K + , H2O2; the A-P probe is obtained by hybridization of an aptamer Apt and a primer P; the nucleotide sequences of the aptamer Apt, the primer P, the capture probe CP and the circular template CT are shown as SEQ ID NO: 1-4; the 5' end of the capture probe CP is modified with SH. The sensor has a simple preparation method, stable performance and is suitable for the detection of OTA in food and environment; the process cost of the preparation process is low, the performance is stable, the electrode has good repeatability and is suitable for the requirements of cheapness in industrialization.

The application provides a 5-aminoacetylpropionic acid-producing strain and a construction method and application thereof. The strain takes Escherichia coli as a chassis, up-regulates the expression amount of a key gene hemA of a 5-aminoacetylpropionic acid synthesis pathway, can improve the carbon flux for synthesizing 5-ALA, and simultaneously inhibits the core gene of a porphyrin precursor synthesis pathway, such as by modifying the RBS and the initiation codon of the hemE gene at the genome level and / or knocking out the hemN gene on the genome, so as to reduce the accumulation of porphyrin intermediates, to realize high-yield 5-ALA, and to effectively avoid the discoloration and deepening of the fermentation broth of the above strain, and to reduce the burden of downstream separation and purification. Further, the 5-aminoacetylpropionic acid dehydrase hemB is weakened, so as to reduce the path of 5-ALA into the porphyrin synthesis pathway, and to improve the 5-ALA yield. Thus, the purpose of improving 5-ALA and avoiding discoloration of the fermentation broth is achieved, and the production cost of 5-ALA is effectively reduced.

This invention relates to an ionic amphiphilic bioporphyrin, its preparation method, and its application, belonging to the field of pharmaceutical technology. The preparation method of the ionic amphiphilic bioporphyrin of this invention includes the following steps: stirring and reacting heme with an alkaline solution to obtain a dispersion; adding an acidic solution to the obtained dispersion to adjust the pH to 4-11, thereby obtaining the ionic amphiphilic bioporphyrin. This invention combines the ionized modified amphiphilic bioporphyrin with alkaline excipients and prepares it into an enteric-coated capsule dosage form, ensuring its stability in gastric acid. Upon reaching the intestine, it achieves effective release and creates a suitable alkaline microenvironment for the dissolution of ionic heme, thereby improving the solubility of heme derivatives in the intestine and oral uptake efficiency.

The application relates to a rapamycin-G4H nanocapsule, a preparation method and application thereof. The preparation method of the rapamycin-G4H nanocapsule comprises the following steps: S1. preparing a rapamycin micelle by adopting a film hydration method; S2. modifying a dibenzocyclooctyne amplification primer on the surface of the rapamycin micelle through a click chemistry reaction to obtain a rapamycin micelle modified with the amplification primer; S3. taking a linear DNA molecule as a template, and synthesizing a circular amplification template through an intramolecular cyclization reaction; and S4. incubating the circular amplification template, the rapamycin micelle modified with the amplification primer, a thermophilic DNA polymerase, a pyrophosphatase, dNTP and Co-G4-haem DNAzyme under the condition of a thermophilic DNA polymerase reaction buffer. The rapamycin-G4H nanocapsule can play an efficient antioxidant stress and immune inflammatory regulation function, can improve the intraocular microenvironment of a glaucoma patient, and can inhibit the continuous apoptosis of retinal ganglion cells.

The invention discloses yarrowia lipolytica genetically engineered bacteria for producing lutein as well as a construction method and application of the yarrowia lipolytica genetically engineered bacteria. According to the invention, the Cu < 2 + > induced promoter pMT2 is introduced, and the lycopene is differentially regulated on the time scale to carry out epsilon-cyclization reaction and beta-cyclization reaction, so that the content of by-product beta-carotene is greatly reduced; by introducing a cipB scaffold protein strategy, a high-efficiency electron transport protein complex for converting alpha-carotene into xanthophyll is constructed, and the content of an intermediate product alpha-cryptoxanthin is effectively reduced; by introducing a synthetic route of cytochrome P450 monooxygenase cofactor heme, the activity of a key catalytic enzyme is improved, and the yield of xanthophyll is effectively improved. The Yarrowia lipolytica engineering strain can utilize common carbon sources and nitrogen sources, the shake flask yield of xanthophyll can reach 93 mg / L after fermentation culture, and the Yarrowia lipolytica engineering strain is an excellent strain with high xanthophyll yield and has great industrial application prospects.

The present invention relates to a transfer platform for a biological sample analyzer, comprising a movable carrier plate and an object stage which is placed on the carrier plate and separable from the carrier plate. The side of the carrier plate facing the object stage is an upper surface, and the reverse side is a lower surface. The side of the object stage facing the carrier plate is a lower surface, and the reverse side is an upper surface. The object stage and the carrier plate are respectively provided with magnetic blocks that magnetically attract each other. When the object stage is placed in a predetermined area on the carrier plate, the suction of the magnetic blocks automatically positions the object stage relative to the carrier plate, thereby achieving the technical effect of blind positioning. The present invention can be applied to test analyzers for specific proteins, cholesterol, heme, routine urine test, dry biochemical test, etc. The present invention has the advantages of convenient operation, time and effort saving, high efficiency and the like. Meanwhile, the present invention is simple in structure and low in cost, and is suitable for wide promotion.

The invention discloses a bacillus amyloliquefaciens riboswitch regulation and control system and application thereof. The regulation and control system is composed of a P43 promoter, a theophylline aptamer TC, RBS, RepA-tag and a required regulation and control gene. A theophylline ribose switch regulation element is integrated on the upstream of a bacillus amyloliquefaciens regulation gene, and the expression quantity of the gene is precisely regulated through theophylline induced expression. Furthermore, a theophylline ribose switch regulation element is integrated to the upstream of the genes hemX and mtnN to obtain the dual-regulation engineering bacterium HZ-B. Under induction of 2mM theophylline, the heme yield and the 5-adenosylmethionine (SAM) yield of the dual-regulation engineering bacterium HZ-B are remarkably increased and are respectively increased by 33.1 times and 0.5 times compared with those of an original strain HZ-12.

The invention belongs to the field of electrochemical sensors, and provides a construction method and application of an electrochemical sensor for detecting the pH of brain glioma, and the specific process is as follows: firstly, a bimetallic MOF (Zr-TCPP (Fe)) with a heme-like structure (containing Fe element and porphyrin ring) is synthesized, and the material has electrochemical response to pH; 12 sulfydryl dodecyl phosphoric acid (12MA) is fixed on the surface of the gold electrode through an Au-S bond. Furthermore, the MOF material is immobilized on the surface of the electrode by using a Zr-O-P bond between 12MA and Zr-TCPP (Fe). The method can be applied to pH detection of the brain glioma tissue extracting solution.

The application belongs to the technical field of sensors, and provides a biosensor for detecting Escherichia coli, which comprises AuNRs modified with H1 hairpin and H2 hairpin, a hairpin probe HAP, an S-T composite probe, hematin, potassium ions, H2O2 and tetramethyl benzidine; the nucleotide sequences of the hairpin probe HAP, the H1 hairpin and the H2 hairpin are shown in SEQ ID NO:1-3; the S-T composite probe is formed by complementary pairing of an S chain and a T chain with the nucleotide sequences shown in SEQ ID NO:4-5; and the H1 hairpin and the H2 hairpin are electrostatically adsorbed on the surface of the AuNRs. The biosensor provided by the application has high reaction efficiency, good specificity and low detection limit, and is suitable for qualitative and quantitative determination of the number of Escherichia coli.

The invention relates to the technical field of biological medicine, in particular to a nano tube based on fluorescently-labeled small peptide and application of the nano tube, the nano tube is a self-assembled nano tube and is specifically formed by electrostatic adsorption of FAM / heme-KD oligopeptide and a KL-7 polypeptide nano tube; the FAM / heme-KD polypeptide is an FAM / heme-KLVFFAED-NH2 peptide fragment, and the KL-7 polypeptide is an Ac-KLVFFAL-NH2 peptide fragment, and the FAM / heme-KD polypeptide is a KLVFFAED-NH2 peptide fragment. The nanotube based on the fluorescently-labeled small peptide shows excellent sensitivity, rapid response capability, specific recognition and affinity to the Abeta40 aggregate, and can be used for detecting the Abeta40 aggregate, detecting fibrosis of the Abeta40 aggregate, screening an Abeta40 aggregate inhibitor and diagnosing Alzheimer's disease.

The present application relates to a kind of DNA / metal organic framework composite nano-enzyme and its preparation method and application, it relates to microneedle technical field.A kind of preparation method of DNA / metal organic framework composite nano-enzyme, the composite nano-enzyme is obtained by incubating metal oxide modified metal organic framework and heme / G-quadruplex DNA together.The present application also provides a kind of microneedle detection system based on the composite nano-enzyme, the detection system includes the composite nano-enzyme and the hydrogel microneedle for extracting skin interstitial fluid.The method extracts human skin interstitial fluid by hydrogel microneedle patch, and the biomarker uric acid in interstitial fluid is detected by the aid of composite nano-enzyme colorimetry.This means of detecting uric acid is compared with the method of detecting uric acid in blood, minimally invasive painless, simple and fast, intuitive and accurate.