18 results about "Haem Oxygenase" patented technology

The invention discloses a heme oxygenase mutant and application of the heme oxygenase mutant in preparation of bilirubin through multienzyme coupling, and belongs to the field of biological catalysis engineering. The forward mutant of the heme oxygenase is obtained through a directed evolution technology, and heme can be efficiently catalyzed to synthesize biliverdin; the bilirubin is synthesized by further introducing biliverdin reductase, coupling cytochrome P450 reductase and formate dehydrogenase, and coupling the two core enzymes with foreign assistant enzymes to construct recombinant escherichia coli to catalyze heme to synthesize bilirubin. In the system, NADPH does not need to be exogenously added, the reaction time is about 4 h, the yield of bilirubin reaches 2.10 g / L, accumulation of intermediate products is not observed in the catalysis process, and the highest yield and the shortest reaction time in bilirubin synthesis reported at home and abroad at present are achieved. Compared with the traditional pig bile extraction, the method disclosed by the invention breaks through the resource limitation, is simple to operate and high in yield, and realizes efficient and green synthesis of bilirubin.

A genetically modified yeast cell, wherein the yeast cell comprises a genetic modification comprising overexpression of yeast gene encoding porphobilinogen deaminase (HEM3), the HEM3 gene having at least 80% identity with SEQ ID No. 7. The genome of the modified yeast cell further comprises one or more genetic modifications in one or more genes selected from: genes coding for heme-dependent repressor of hypoxic genes (ROX1), genes coding for heme oxygenase (HMX1), genes coding for a receptor for vacuolar proteases (VPS10), and genes coding for vacuolar proteinase (PEP4), the one or more genetic modifications being such that expression of a polypeptide from such a gene is reduced or disrupted or the polypeptide expressed is non-functional.

1, 4-disubstituted-1, 2, 3 triazole compounds of formula (I): (I) capable of inhibiting heme oxygenase activity, their use as a medicament for the treatment and / or prophylaxis of pathological conditions associated with alteration of heme oxygenase activity and pharmaceutical compositions comprising the same.

The invention relates to the technical field of bioengineering bacteria. The invention provides a saccharomyces cerevisiae engineering bacterium capable of synthesizing p-anisaldehyde from the beginning and a construction method of the engineering bacterium. A saccharomyces cerevisiae recombinant strain JS1-15 is used as an original strain; the saccharomyces cerevisiae engineering bacteria are obtained by heterologous expression of 4-hydroxymandelic acid synthetase HmaS, heme oxygenase HMO, benzoyl formate decarboxylase BFD and methyltransferase 4VPMT1. The saccharomyces cerevisiae engineering bacteria are used for preparing the saccharomyces cerevisiae engineering bacteria. The saccharomyces cerevisiae engineering bacterium disclosed by the invention can realize a complete de novo biosynthesis path from 4-hydroxyphenylpyruvic acid to p-anisaldehyde by taking glucose as a unique carbon source, and has good industrial application potential.

The present invention discloses a method for promoting heart valve endothelialization and an artificial heart valve. The method is based on modifying circular DNA on the surface of a decellularized heart valve (DHV). A DNA hydrogel targeting thrombin is obtained on the surface of the DHV by rolling circle amplification. The DNA hydrogel not only exerts an anticoagulant function, but also acts as a heme capturer, recruiting heme from the blood. The heme is further converted into biliverdin (BV) by the heme oxygenase (HO1) loaded in the DNA, ensuring the continuous production of biliverdin, promoting anti-inflammatory immune regulation and reactive oxygen scavenging, thereby creating a regenerative immune microenvironment and promoting valve endothelialization.

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.

Escherichia coli is a widely studied model organism and an integral component of the human gut microbiome, offering significant potential for bacteria-based therapeutic applications. However, engineering native E. coli strains poses persistent challenges. In this study, the chassis- independent recombinase-assisted genome engineering technique was leveraged to engineer the native gut strain E. coli EcAZ-1 and the probiotic strain E. coli Nissle 1917 (EcN). The bioluminescent lux operon, green fluorescent protein (GFP), and the oxygen-independent fluorescent protein IFP 2.0 were successfully introduced into both strains. To further enhance IFP 2.0 fluorescence, a heme oxygenase was co-expressed, and the chromophore biliverdin was supplemented, achieving robust IFP 2.0 expression under both anaerobic and aerobic conditions. Also, both strains were engineered to biosynthesize bioactive compounds, including the plant- derived flavonoid naringenin and mycosporine-like amino acids. The results underscore the potential of native E. coli strains as flexible and robust platforms for synthetic biology, enabling novel applications in biomedical research and therapeutics. Based on these results, this application provides recombinant bacteria generating IFP2.0 fluorescence under anaerobic condition, bacterial composition for gastrointestinal cancer detection and use method thereof, and naringenin or MAA overproducing recombinant bacteria.

Methods for identifying chronic hydrocephalus include assaying for an amount in a biological sample of one or more biomarkers selected from pyruvate dehydrogenase lipoamide kinase isozyme 4 (PDK4), phosphofructokinase-muscle (PFKM), low density lipoprotein receptor adaptor protein 1 (LDLRAP1), glucagon-like peptide-1 receptor (GLP-1R), hemoglobin subunit alpha 1 (HBA1), hemoglobin subunit alpha 2 (HBA2), heme oxygenase 1 (HMOX1), and combinations thereof. Methods for screening for a compound useful for treating chronic hydrocephalus are also provided and include contacting a cell with an effective amount of a test compound, and then detecting an expression or activity level of one or more of the biomarkers.

The present application relates to sewage treatment technical field, specifically relates to a kind of cultivation method of anaerobic ammonia oxidation granular sludge resistant to high concentration nano Fe3O4.The specific technical scheme is: by using gradient acclimation+control final concentration+pH down-regulation mode directional stimulation hematin oxygenase gene, realize the stable operation of anaerobic ammonia oxidation system under the existence of long-term high concentration nano Fe3O4.On the one hand, anaerobic ammonia oxidation bacteria is rich in hematin characteristics, provides sufficient substrate for hematin oxygenase-mediated hematin degradation metabolism;On the other hand, using the characteristics of Fe3O4, it releases Fe (II) and Fe (III) under low pH condition.The present application solves the problem of long-term growth inhibition of high concentration nano Fe3O4 on Anammox bacteria, and creates the possibility for the practical application and popularization of Anammox system in treating wastewater containing nano materials.

The invention discloses escherichia coli engineering bacteria with high yield of phycocyanobilin as well as a construction method and application of the escherichia coli engineering bacteria, and belongs to the technical field of synthetic biology. According to the engineering bacteria, escherichia coli BL21 (DE3) is taken as a host, heme oxygenase (HO) which is optimized by codons and is derived from Cornebacterium diphtheria and phycocyanobilin reductase (PcyA) which is derived from Synecocystis sp.PCC6803 are expressed, and a trxA dissolution promoting label is fused, so that the solubility and the stability of zymoprotein are effectively improved; meanwhile, a high-copy plasmid pRSFDuet is adopted for enhancing gene expression, a heme synthetase gene hemH, HO and PcyA are used for constructing a multi-enzyme complex through connecting peptides, precursor supply and enzyme space coordination are enhanced, and the synthesis efficiency of phycocyanobilin is remarkably improved. The yield of phycocyanobilin reaches up to 37 mg / L in shake flask fermentation, and the engineering bacterium is simple in process, low in cost and suitable for large-scale production.

The application discloses a kind of heme oxygenase mutant and its multienzyme coupling preparation of application of bilirubin, belong to biological catalysis engineering field.The application obtains the positive mutant of heme oxygenase by directed evolution technology, can efficiently catalyze heme to synthesize chlorophyllin;Further introduce chlorophyllin reductase, coupling cytochrome P450 reductase, formic acid dehydrogenase, and two kinds of core enzyme are coupled with exogenous auxiliary enzyme, constructs recombinant escherichia coli, catalyzes heme to synthesize bilirubin.In the system, without exogenous addition NADPH, reaction time is about 4h, and bilirubin yield is as high as 2.10g / L, and no intermediate accumulation is observed in catalytic process, reaches the highest yield of the synthesis of bilirubin reported at home and abroad, and the shortest reaction time level.Compared with traditional pig bile extraction, the method of the application breaks through the resource limit, and the operation is simple, the yield is high, realizes the efficient, green synthesis of bilirubin.

Disclosed is a method for producing phycocyanobilin by use of a recombinant Escherichia coli that express heterologous heme oxygenase ho1 and ferredoxin oxidoreductase pcyA derived from Synechocystis sp. PCC6803. According to the present disclosure, heterologous expression of ho1 and pcyA genes leads to conversion of heme to an intermediate biliverdin for phycocyanobilin synthesis, and reduces the accumulation of biliverdin in the process of the phycocyanobilin synthesis. The genome of E. coli is further engineered to overexpress related genes of a metabolic pathway of phycocyanobilin, and a strain of recombinant E. coli with high yield of phycocyanobilin is obtained. The recombinant E. coli strain is cultured for 36 hr in a system using glycerol as a substrate, and the phycocyanobilin yield can reach 147 mg / L.

The application discloses an escherichia coli zjut-mhol and application thereof in production of heme oxygenase-1, and a gene of mouse-derived HO-1 is synthesized and cloned by using a preferred codon optimization method, a gene recombinant escherichia coli capable of producing HO-1 is constructed, and a mutant strain with high enzyme activity and stable passage is obtained through ultraviolet mutagenesis, and the mutant strain can be applied to preparation of HO-1. The strain is cultured for enzyme production, and the bacterial body with intracellular active HO-1 is obtained, and the enzyme production activity is 122 U / g according to the mass of the dry bacterial body. The HO-1 freeze-dried powder is separated from the bacterial body, and the enzyme activity of the freeze-dried powder is 91.8 U / g.

The invention belongs to the technical field of biological medicines, and particularly relates to a tetrahedral framework nucleic acid combined with siRNA related to a silent NF-kappa Bp65 gene as well as a preparation method and application of the tetrahedral framework nucleic acid. The invention combines the strong cell membrane permeability of tetrahedral framework nucleic acid and the gene targeting specificity of siRNA-NF-kappa Bp65, and provides a tetrahedral framework nucleic acid system combined with siRNA related to silent NF-kappa Bp65 gene, and the system can effectively and continuously down-regulate the expression of NF-kappa Bp65, so that the expression of the siRNA related to silent NF-kappa Bp65 can be effectively and continuously regulated by inhibiting Wnt / beta-catenin, and the expression of siRNA related to silent NF-kappa Bp65 can be effectively and continuously regulated. And the NRF2 / heme oxygenase-1 (HO-1) pathway is up-regulated to inhibit the joint inflammation and promote the repair of the articular cartilage. The delivery system provided by the invention has a more excellent treatment effect in treatment of temporomandibular arthritis, and is expected to become a new way for treatment of temporomandibular arthritis.

The present application relates to the technical field of NK cell culture, and discloses a method for improving the killing function of NK cells and application thereof.The method comprises the following steps: (1) adding a haem oxygenase 1 antibody to an NK cell culture medium to prepare an antibody culture medium; (2) performing sterile filtration treatment on the antibody culture medium; and (3) separating NK cells and culturing the NK cells in the antibody culture medium.The present application adds a haem oxygenase 1 antibody to treat NK cells, blocks the expression of haem oxygenase 1 in the NK cells, or inhibits the activity of an effective substance of haem oxygenase 1.After being treated with the haem oxygenase 1 antibody, the secretion of granzyme B and perforin of the NK cells can be promoted, and the killing effect of the NK cells on target cells Yac-1 and K562 can be improved, so that the anti-tumor function of the NK cells is improved.

The invention discloses a preparation method and application of a dendritic cell-derived exosome, and the preparation method comprises the following steps: (1) inducing in vitro differentiation of bone marrow-derived dendritic cells (DCs) to tolerant dendritic cells (tolDCs) through interleukin-10, and ensuring that the tolDCs highly express key antioxidant and anti-inflammatory proteins, namely, heme oxygenase-1 (HO-1); (2) separating and purifying an exosome HO-1 high-tolDex from the cell culture supernatant of the tolDCs (HO-1 high-tolDCs) with high expression of the HO-1 by virtue of methods such as ultracentrifugation and the like; (3) carrying out comprehensive physicochemical property and biological function identification on the prepared HO-1high-tolDex, and (4) verifying that the HO-1high-tolDex can effectively inhibit excessive activation and inflammatory response of target cells through an in-vitro cell experiment and an in-vivo experiment of a systemic lupus erythematosus (SLE) animal model, alleviate immune pathological injury of model animals, inflammation of kidneys and oxidative stress injury, and improve the bioavailability of the model animals. The immune tolerance is induced; the cell-free immunomodulator is a new generation of efficient and safe cell-free immunomodulator.

The present invention provides, among other things, methods and compositions for making and using recombinant heme oxygenase for treating sickle cell disease. In some embodiments, recombinant heme oxygenase proteins are truncation variants, or Fc fusion proteins with increased half-life and / or reduced aggregation.

The invention discloses a preparation method and application of taurine carbon dots. Comprising the following steps: S1, putting a carbon source, a nitrogen source and an active dopant into a solvent, and fully mixing and dissolving to obtain a mixed solution; s2, heating the mixed solution by microwaves to obtain a final product. The mass ratio of the carbon source to the nitrogen source to the active dopant is (4.5-5.5): (4-6): (1-3). And S2, the microwave radiation time is 4 to 5 minutes. The taurine carbon dots are synthesized by doping a carbon source, a nitrogen source and an active dopant, the influence on the activity, proliferation, migration, invasion, adhesion, clone formation ability, cell cycle arrest and apoptosis of a CRC cell line shows that Tauu / CDs significantly inhibits tumor growth, and a basis is provided for treatment and inhibition of Tauu / CDs in CRC; through analysis and ferroptosis evaluation, the close relation between Taus / CDs treatment and ferroptosis activation mediated by heme oxygenase 1 is revealed, and an important theoretical basis is provided for pharmaceutical preparations.