92 results about "Heme" patented technology

The invention belongs to the technical field of plant genetic engineering, and particularly relates to HBP3 for improving plant salt tolerance and application of the HBP3, the nucleotide sequence of the HBP3 is shown as SEQ ID NO.3, and the amino acid sequence of the HBP3 is shown as SEQ ID NO.4. According to the invention, a gene HBP3 for pre-coding the heme binding protein is separated from red rice for the first time, and an overexpression plant is obtained by constructing a recombinant overexpression vector containing the HBP3 and transforming the recombinant overexpression vector into an arabidopsis plant. Compared with a wild type plant, the over-expression plant has the advantage that the tolerance to high salinity is enhanced. According to the invention, the salt stress resistance of HBP3 is found for the first time, the salt stress experiment result shows that the high salt tolerance of the overexpressed plant is obviously enhanced, and the expression of the two genes ABA1 and ABA2 related to abscisic acid in the overexpressed plant is reduced, so that a molecular basis is provided for revealing the salt stress regulation of the overexpressed plant.

The invention discloses a lung adenocarcinoma prognosis prediction model based on heme metabolism related genes and a construction method and application thereof. According to the invention, a heme metabolism related gene set is screened through a molecular characteristic database, and a gene significantly related to prognosis is screened by using transcriptome and clinical data of lung adenocarcinoma in a TCGA database and combining single-factor Cox regression, LASSO regression and multivariable Cox analysis. And through multi-time iterative modeling, selecting high-frequency stable genes and corresponding regression coefficient mean values to construct a heme metabolism risk score. According to the method, a random survival forest model is adopted to carry out iteration evaluation on gene importance for multiple times, the first 50% of genes which have the maximum influence on the model are screened, and finally the five important characteristic genes related to the heme metabolism risk are identified by taking an intersection with an LASSO result. The application provides new theoretical basis and technical support for precise treatment of lung adenocarcinoma, and has important clinical application value.

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).

Provided herein are methods of functionalizing C(sp3)-H bonds using reprogramed metalloenzymes to perform radical-relay C(sp3)-H functionalization, activating a (sp3)-H bond via a reactive radical (X·) via hydrogen atom transfer (HAT); intercepting of the resulting carbon-centered radical by a redox-reactive metal complex; and obtaining a functionalized C—Y bond.

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 the technical field of heme processing and utilization, and particularly discloses a method for efficiently dissolving heme. The preparation method comprises the following steps: mixing a hydrogen bond donor and a hydrogen bond acceptor according to a molar ratio of 1: 0.5-1: 8, heating in a water bath to prepare a deep-eutectic solvent, adding heme into the prepared deep-eutectic solvent, and fully and uniformly mixing the heme and the deep-eutectic solvent according to a volume ratio of 1-3.5 mg / ml. The method is simple, the steps are easy to operate, by adopting the method, the solubility of the heme is improved, the stability of the heme can be maintained, and a foundation is laid for further development and utilization of the heme.

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.

Indoleamine 2,3-dioxygenase (IDO) is a heme-binding enzyme involved in pathophysiological processes including, but not limited to, antimicrobial and antitumor defense, neuropathology, immunoregulation, and antioxidant activity. Aspects of the disclosure relate to variants of IDO that comprise one or more amino acid variations relative to a wild-type IDO and / or that are connected to a synthetic polymer, such as poly (ethylene glycol). Embodiments of the disclosure provide variant IDOs that have improved properties, such as resistance to oxidative conditions, protein solubility, lower propensity to aggregate in vivo, increased half-life in the circulatory system upon administration to a subject, increased binding to a cell or tissue, or any combination thereof. Accordingly, IDO variants of the disclosure can be administered to a subject in need thereof, such as a subject having, suspected of having, or at risk of developing a disease, disorder, or condition, such as one associated with inflammation.

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 invention discloses a biosensor for detecting heme, hemoglobin and / or L-cysteine ​​and its application, belonging to the field of biosensing and analysis. The biosensor comprises a hairpin nucleic acid HP, a binding buffer solution, heme, TMB, H2O2 and a HAc-NaAc buffer solution; the sequence of the hairpin nucleic acid HP is shown in SEQ No. 1, and its 5' end is labeled with a quenching group BHQ1, and its 3' end is labeled with a fluorescent group FAM. The present invention can realize the optical detection of heme and hemoglobin in the "turn-on" mode and the rapid detection of L-cysteine. It has higher sensitivity than existing detection methods, and experiments have shown that it is still applicable in complex serum systems. It is expected to provide a simple, rapid and sensitive method for the detection of heme, hemoglobin and L-cysteine ​​in actual clinical samples.

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*.

The invention discloses a solid-state self-enhanced ECL aptamer thrombin sensor as well as a construction method and application of the solid-state self-enhanced ECL aptamer thrombin sensor. The method comprises the following steps: (1) synthesizing conjugated polymer points PDC; (2) incubating a PDC / dicarboxyl bipyridine ruthenium solid self-enhanced ECL working electrode; and (3) construction of the conjugate heme-G-quadruplex / AuNPs (AuNPs). The branched chain polyethyleneimine and tri-n-propylamine are similar in multi-tertiary amine structure, so that the branched chain polyethyleneimine and a polymer point PDC formed by hydrothermal synthesis of NH2BDC also have the capability of serving as a bipyridyl ruthenium co-reaction reagent, the detection range of the ultra-sensitive TB aptamer sensor of a sandwich structure constructed based on the PDC is 1015-108 M, the fluctuation range of TB in a blood sample is included, and the sensitivity of the TB aptamer sensor is improved. The detection limit is 3.6 * 10 <-16 > M, and the probe has an application prospect in thrombin medical detection.

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.