33 results about "Heme" patented technology

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.

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.

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 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.

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.

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.

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 application discloses a TteUPO mutant and application thereof, and belongs to the technical field of bioengineering and enzyme engineering.The TteUPO mutant disclosed by the application connects the nucleotide sequence of a short-chain non-specific peroxidase with a SUMO and 8HIS tag to a pET28a vector, obtains a recombinant vector, transforms E.coli BL21 (DE3), and obtains a genetically engineered bacterium; and the mutant is obtained through unit point mutation.The application uses an E.coli autoinduction system, can make shake flask culture grow to high density in a short time, contains rich nutrients in the culture medium, and does not need to monitor cell density and add IPTG in the expression process.Meanwhile, a hematin precursor 5ALA is added in the E.coli autoinduction culture medium, and finally, a high-activity short-chain non-specific peroxidase is obtained, and the enzyme after purification is reacted with anisole under the condition of hydrogen peroxide flow addition to obtain a product.

The invention belongs to the technical field of biological detection, and particularly relates to a sample treating fluid for detecting bordetella pertussis, a detection method, a product and application. The sample treatment liquid comprises a lysis working solution (WLB) and a post-treatment diluent (PD), the WLB is composed of choline tartrate, betaine, glycerin, TCEP, IGEPAL CA-630, HEPES and non-enzyme deionized water, and the PD is composed of bovine serum albumin and non-enzyme deionized water. The kit is suitable for common clinical samples such as sputum and nasopharynx swab eluent, bordetella pertussis can be efficiently split, target nucleic acid is fully released, and meanwhile matrix inhibitors such as mucoprotein, heme and protease in the samples are effectively removed. The kit is simple and convenient to operate, free of complex purification steps, high in detection sensitivity, good in reproducibility, high in specificity and free of cross reaction, and provides reliable technical support for early precise diagnosis and epidemic prevention and control of pertussis.

The present invention is characterized by compositions and methods for editing harmful mutations associated with heme disorders such as sickle cell disease (SCD). In a particular embodiment, the present invention provides a method for correcting mutations in β-globin polynucleotides using a modified adenosine base editor called “ABE8” with unprecedented levels of efficiency (e.g., >60 to 70%).

The invention provides a kit for detecting tumor cells based on electrochemical analysis and application of the kit, and belongs to the technical field of biosensing. The invention discloses a kit for detecting tumor cells based on electrochemical analysis. The kit comprises an ordered functionalized electrode, a recognition catalytic probe, heme, tyramine alkyne, hydrogen peroxide, a double-chain signal probe modified with an azide group and an electrochemical active group, and a solution containing cuprous ions. According to the kit, a double-target collaborative tumor cell recognition strategy combining a small molecule ligand and a nucleic acid aptamer is adopted, so that the selectivity and the detection specificity of target tumor cells are remarkably improved; by virtue of an ordered functionalized electrode interface, the capture efficiency of target tumor cells is effectively improved, and the interface adsorption of non-target components is reduced; in addition, through the synergistic signal amplification effect of proximity oxidation labeling and click chemistry, background interference caused by non-target cells is effectively inhibited while high-sensitivity detection is achieved, and the probe still has excellent detection performance in complex biological samples.

The present disclosure provides methods to detect a target in a sample by using a DNA scaffold with enzymatic activity (“DNAzyme”) that catalyzes hydrogen peroxidase decomposition, leading to a reaction wherein bubbles are produced. In particular, the present disclosure describes the capabilities of G-quadruplex·hemin constructs to produce bubbles as a signal transduction tool in biomarker detection. Observing bubbles is a straightforward and easy-to-interpret route towards instrument-free visualization of readouts that does not require a scientific background, an equipped lab, or a color vision proficiency.

This invention provides a hydantoin mutant, related products, and their application in the production of the pharmaceutical intermediate (R)-3-(carbamoylmethyl)-5-methylhexanoic acid or its analogues and downstream products. Compared with wild-type hydantoin, the hydantoin mutant of this invention exhibits significantly enhanced catalytic activity, and can stably maintain a high enantiomeric excess (ee value) during the catalytic synthesis of (R)-3-(carbamoylmethyl)-5-methylhexanoic acid. Therefore, using the hydantoin mutant of this invention, high-purity R-3-(carbamoylmethyl)-5-methylhexanoic acid or its analogues can be produced in a low-cost and high-efficiency manner, thus possessing outstanding application value and broad industrialization prospects.

The invention relates to a preparation method and application of a fluorescent colorimetric electrochemical three-mode sensor based on G-quadruplex combined hyperbranched HCR. A large number of DNA branched chains are constructed by adopting a hyperbranched HCR signal amplification strategy, compared with a traditional HCR technology, hyperbranched HCR provides more sites for embedding of signal tags, signal amplification is achieved, and cost is reduced. The constructed dendritic DNA branched chain combined G-quadruplex plays a role of peroxidase by adding heme, can play a role of catalyzing 3, 3 ', 5, 5'-tetramethyl benzidine (TMB) by catalyzing hydrogen peroxide so as to play a role of colorimetry, and in addition, the dendritic DNA branched chain can be embedded with a large number of electrochemical and fluorescent signal labels, so that the purpose of multi-mode detection is achieved. The prepared three-mode sensor shows relatively high sensitivity, relatively wide detection range, good selectivity and excellent practical application capability.

Provided is a method for measuring heme pigments in meat more easily than in the past without using special equipment.　Heme pigments are extracted from meat to obtain an extract, and the post-oxidation extract is dispensed to obtain a first extract and a second extract. The pH of the first extract is adjusted to 5.0-7.5 and the pH of the second extract is adjusted to 8.0-10.0, the extracts are oxidized, and measurements are taken of a first absorbance, which is the absorbance of the first extract at a predetermined wavelength, and a second absorbance, which is the absorbance of the second extract. The myoglobin concentration and the hemoglobin concentration in the meat are calculated from the difference between the measurements taken for the first absorbance and second absorbance, whereby the heme pigments in the meat is measured.

The present invention relates to a recombinant heme or phytoglobin protein comprising C-helices and D-helices covalently linked to a CD loop region, and comprising a hexa-coordinate or penta-coordinate prosthetic heme group, wherein the heme or phytoglobin has an amino acid sequence that is at least 50% identical to the heme or phytoglobin contained in any one of SEQ ID NOs. 1 to 55, the CD loop region comprises one or more modifications compared to the parent CD loop region, thereby the modified CD loop region is more strongly bound to the prosthetic group than the parent unmodified CD loop region, the parent CD loop region is located at positions 52 to 78 of SEQ ID NOs. 1 in the recombinant heme or phytoglobin protein, optionally at positions 52 to 72 of SEQ ID NOs. The CD loop region has an amino acid sequence that is at least 50% identical to the CD loop region contained in any one of SEQ ID NOs. 56 to 147.

The application provides a hemoglobin cascade response nano vesicle and a preparation method and application thereof, and belongs to the technical field of biological medicine. The first hemoglobin response nano vesicle (DecAS-PC@NM) is designed, and disease-specific treatment is realized through cascade reaction of two functional phospholipids. First, hemoglobin activates artemisinin-modified phospholipid (A-PC) to generate active oxygen, the active oxygen oxidizes the sulfide structure in another phospholipid (S-PC), and the transformation from hydrophilicity to hydrophobicity is caused, so that the vesicle is disintegrated. Through co-assembly with neutrophil membranes, the chemotaxis to inflammatory sites is further enhanced. In vitro studies show that DecAS-PC@NM has unique hemoglobin responsiveness, and in vivo data emphasize its targeting to ischemic cores and significant therapeutic effect. This innovative hemoglobin-triggered phospholipid cascade reaction provides a promising strategy for the treatment of myocardial ischemia-reperfusion injury.