10 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.

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 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.

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 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 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 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.