115 results about "Heme" patented technology

The invention discloses recombinant escherichia coli for producing heme and derivative compounds thereof and a preparation method of the recombinant escherichia coli. According to the invention, on the basis of escherichia coli, ptsG, galR, lpxM, pta, poxB, arcA, ldhA, sdhA, iclR, metA and dapD are knocked out; the method comprises the following steps: introducing glk, Zglf, acs, ppc, BsmetA, BsyjcI, BsdapH, BspatA and BscdapL, and enhancing the expression of araE, stA, coA, serA and gltA; and the expression of icd is weakened. And the expression of eamA and rhtA is enhanced. The GYDH and the hemA are introduced; according to the recombinant strain, D-glucose is used as a raw material to synthesize 5-ALA, and on the basis of the 5-ALA, coding genes of the heme and the various derivative compounds thereof are introduced, so that recombinant bacteria for producing the heme and the various derivative compounds thereof can be obtained.

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 present invention relates to methods for producing heme comprising culturing a microorganism that secretes coproporphyrin III and / or coproheme III in a culture medium and converting the secreted coproporphyrin III and / or coproheme III into heme, as well as to genetically engineered microorganisms capable of secreting uroporphyrin III, uroporphyrin I, coproporphyrin III and / or coproheme III.

The invention relates to the discovery of a biosynthetic gene cluster of an alkaloid, namely, exopoid C, and the biosynthetic gene cluster of the exopoid. A biosynthetic gene cluster exo of an exopoid compound is obtained from a fungus Penicillium sp.HDN14-431 through genome mining, the gene cluster comprises a heme-dependent oxidase gene exoA which is composed of indole dioxygenase and a cytochrome b5 structural domain, a cytochrome P450 enzyme gene exoB and an isopentenyl transferase gene exoC which is composed of indole dioxygenase and a cytochrome b5 structural domain, and the cytochrome P450 enzyme gene exoB and the isopentenyl transferase gene exoC are used as biosynthetic genes of the exopoid compound. All genes in a gene cluster are expressed through an aspergillus nidulans heterologous expression system, two isopentenyl indole alkaloids are obtained in total, and a compound I is a new compound exopoid C. And the compound II, namely the exopoid, is a known compound. The biosynthesis pathway of the compound is reported for the first time.

The invention discloses a Taq DNA polymerase mutant with improved thermal stability and inhibition resistance as well as a preparation method and application of the Taq DNA polymerase mutant, and relates to the technical field of biology. According to the Taq DNA polymerase mutant, amino acid mutation is carried out on an amino acid sequence of wild type Taq DNA polymerase, and amino acid mutation sites comprise F73S and P93A; the amino acid sequence of the wild type Taq DNA polymerase is as shown in SEQ ID NO: 1. Compared with wild type Taq DNA polymerase, the Taq DNA polymerase has the advantages that the resistance to inhibitors such as heme, ethanol and EDTA (Ethylene Diamine Tetraacetic Acid) is improved, and meanwhile, the Taq DNA polymerase has higher specific enzyme activity and heat stability.

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 discloses a method for preparing nitrone compounds by using a non-heme double-iron-ion enzyme, and belongs to the field of biological enzyme catalysis. A substituted styrene compound and a substituted aniline compound are used as substrates and react in a PBS buffer solution in the presence of non-heme double iron ion type amino oxidase AzoC to obtain the nitrone compound. According to the present invention, the enzyme catalysis nitrone synthesis system verifies the feasibility of the non-heme double iron ion type amino oxidase mediated bimolecular nitrone compound synthesis, the solid foundation is laid for the subsequent research and application, and the strategy provides the basis for the subsequent enzyme modification and optimization.

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 invention discloses a preparation method of a NM88B-POR / Rf nanozyme and its application. The method first in-situ dopes porphyrin in NM88B(Fe) (NM88B-POR), and then loads riboflavin (Rf) on NM88B-POR by electrodeposition, while realizing the sarcosine oxidase (SOx) nanozyme Rf and peroxidase nanozyme NM88B(Fe) enzyme cascade reaction to enhance the sarcosine (Sar) detection signal, and also enhances the capture of oxygen molecules in aqueous solution through the heme-like effect of the Fe-POR structure in NM88B-POR to construct a high oxygen microenvironment, thereby realizing highly sensitive electrochemical detection of Sar. The present invention establishes a new Sar enzyme cascade electrochemical detection strategy, and the heme-like Fe-POR structure in the constructed electrochemical nanozyme can also build a high-oxygen microenvironment by enhancing the capture of dissolved oxygen, solving the problem that the current Rf nanozyme can only biomimetic the SOx active center structure and ignores the fact that the capture of dissolved oxygen during the reaction process can significantly improve the reaction rate. The nanozyme ultimately achieves ultra-sensitive electrochemical detection of Sar through the synergistic effect of enzyme cascade and high-oxygen microenvironment.

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 discloses a multifunctional cascade nano-enzyme and a preparation method and application thereof, and belongs to the technical field of medical nano-materials, the multifunctional cascade nano-enzyme is a polydopamine nano-material doped with gold nanoparticles and heme, and the surface of the polydopamine nano-material is modified with a targeting ligand or a stabilizer. According to the nano material, gold ions, heme and dopamine hydrochloride are self-assembled under an alkaline condition to form nano particles, and a targeting ligand or a stabilizer is further modified through a chemical reaction. The multifunctional cascade nano-enzyme disclosed by the invention can convert glucose in a disease microenvironment into hydrogen peroxide in situ and can also convert the glucose into toxic hydroxyl radicals, so that the redox balance of tumor cells and bacteria is destroyed, apoptosis and death of the tumor cells and the bacteria are induced, toxic and side effects on normal cells and tissues can be effectively avoided, and the treatment effect is good. And the polymer has certain microenvironment regulation and control capability, so that the polymer can be used for preparing photo-thermal therapeutic agents, drug therapeutic agents and photo-thermal drug combined therapeutic agents for tumors or infected wounds.

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.