41 results about "Proline Hydroxylase" patented technology

The invention discloses a gene segment provided with coded and highly-active trans-4-hydroxyl-L-prolyl hydroxylase and an application thereof. Gene codons of trans-4-hydroxyl-L-prolyl hydroxylase (GenBank ID: BAA 20094.1) in Dactylosporangiumsp. RH1 are optimized to obtain a p4hyd gene; the optimized p4hyd gene is truncated to obtain a P4Hyd1-257 gene; recombinant plasmids including pET-M-3C-P4Hyd and pET-M-3C-P4Hyd1-257 are built respectively, prokaryotic expression is performed through escherichia coli, and trans-4-hydroxyl-L-proline is produced in a conversion manner by utilizing thalli as an enzyme source. An experimental result shows that L-proline is taken as a substrate, compared with pET-M-3C-P4Hyd, the highest conversion rate of trans-4-hydroxyl-L-proline generated by pET-M-3C-P4Hyd1-257 in the conversion manner is increased from 91.4% to 97.4%, the conversion time is shortened from 80 hours to 60 hours, so that the conversion rate is improved remarkably, and the conversion time is shortened remarkably. The gene segment provided with coded and highly-active trans-4-hydroxyl-L-prolyl hydroxylase and the application thereof can be used for producing L-proline by a bioconversion method, and have better industrial application prospect.

The invention discloses a high-efficiency conversion method for producing trans-4-hydroxy-L-proline by a bioconversion technology. The high-efficiency conversion method comprises a high-activity self-induced expression method of L-prolyl hydroxylase in recombinant engineered escherichia coli, types and concentrations of assistants used in an L-proline-to-trans-4-hydroxy-L-proline efficient conversion system, and conversion conditions of the conversion system. The high-efficiency conversion method provides an important scientific basis for establishing bioconversion-based industrial production of trans-4-hydroxy-L-proline.

The invention relates to a 5-hydroxy-1,7-naphthyridine compound substituted by an aryloxy group or a hetero-aryloxy group, a preparation method and a pharmaceutical application thereof. In particular, the compound is represented as the formula (I), wherein R2, R3 independently are hydrogen; R1 is hydrogen or a C1-C3 alkyl group; Ar is an aromatic ring or a hetero-aromatic ring selected from naphthalene ring, pyridine ring, thiophene ring, furan ring and substituted benzene ring. The invention also relates to the preparation method of the compound, a medicine composition including the compound or a pharmaceutically acceptable salt thereof, and an application of the compound or the pharmaceutically acceptable salt thereof in preparation of a medicine for inhibiting HIF proline hydroxylase or a medicine for promoting generation of endogenous EPO.

The invention relates to a dihydropyrazolone compound as well as a preparation method and medical application thereof, in particularly relates to a general formula (I) compound shown in the specification, as well as a preparation method thereof, a pharmaceutical composition containing the compound, and application of the compound as a proline hydroxylase (PHD) inhibitor, the compound and the pharmaceutical composition containing the compound can be used in treating and / or preventing of PHD -activity- related diseases such as cardiovascular diseases, chronic kidney disease, anemia, wounds, cancer, autoimmune diseases and the like. In the general formula (I), the definition of each substituent in the formula is the same as that in the specification.

The invention discloses a novel proline 3-hydroxylase. The proline 3-hydroxylase has an amino acid sequence as shown in SEQ ID NO:2 or SEQ ID NO:4. The invention also discloses an expression vector and a host cell containing the enzyme or a coding gene thereof as well as applications of the expression vector and the host cell in production of trans-3-hydroxy-L-proline. The proline 3-hydroxylase disclosed by the invention has high catalytic performance, so that the trans-3-hydroxy-L-proline can be produced by taking L-proline or glucose as a substrate, the production cost is greatly reduced, and the industrial production of the trans-3-hydroxy-L-proline is further realized.

The invention discloses vegetable protein peptide powder and a preparation method thereof. The method comprises the steps: (1), uniformly mixing vegetable protein powder and water according to the mass ratio, and adjusting the pH value to 7.0-7.5; (2) adding a compound enzyme I, and carrying out heat preservation at the temperature of 40-50 DEG C, wherein the compound enzyme I comprises proline hydroxylase, neutral peptide chain endonuclease and trypsin; (3) after the enzymolysis is finished, heating to 92-98 DEG C, and carrying out heat preservation; (4) after cooling, adjusting the pH valueto 8.0-8.5, then adding a compound enzyme II into the vegetable protein liquid, and carrying out heat preservation at the temperature of 40-50 DEG C, wherein the compound enzyme II comprises plant proteolytic enzyme, papain and flavor enzyme; (5) after the enzymolysis is finished, heating to 92-98 DEG C, and carrying out heat preservation; and (6) after cooling, sequentially carrying out centrifugal separation, membrane separation desalination and freeze drying to obtain the vegetable protein peptide powder. According to the method, the protein peptide powder can be prepared from vegetable protein, and the tryptophan content is high.

The present invention provides engineered proline hydroxylase polypeptides for the production of hydroxylated compounds, polynucleotides encoding the engineered proline hydroxylases, host cells capable of expressing the engineered proline hydroxylases, and methods of using the engineered proline hydroxylases to prepare compounds useful in the production of active pharmaceutical agents.

Biocatalysts and methods for hydroxylation of chemical compounds. The present disclosure provides engineered proline hydroxylase polypeptides for the production of hydroxylated compounds, polynucleotides encoding the engineered proline hydroxylases, host cells capable of expressing the engineered proline hydroxylases, and methods of using the engineered proline hydroxylases to prepare compounds useful in the production of active pharmaceutical agents. The present disclosure provides engineered proline hydroxylase biocatalysts, polynucleotides encoding the biocatalysts. methods of their preparation, and processes for preparing hydroxylated compounds using these engineered biocatalysts.

The present invention provides engineered proline hydroxylase polypeptides for the production of hydroxylated compounds, polynucleotides encoding the engineered proline hydroxylases, host cells capable of expressing the engineered proline hydroxylases, and methods of using the engineered proline hydroxylases to prepare compounds useful in the production of active pharmaceutical agents.

The invention relates to the technical field of biological genetic engineering recombination, in particular to a recombinant human type III collagen mature peptide modified with hydroxyproline and its preparation method and application. The invention provides a gene encoding a recombinant human type III collagen mature peptide, and combines the proline hydroxylase expression vector pPZAStA-P4HA2-P4HB2 with the nucleotides of the gene encoding a recombinant human type III collagen mature peptide The recombinant vector obtained by sequence ligation is transferred into the yeast expression system to obtain a yeast that can express the mature peptide of recombinant human type III collagen modified with hydroxyproline, and use the yeast to express the recombinant human type III modified with hydroxyproline Collagen mature peptide, the recombinant human type III collagen mature peptide has a large molecular weight, a high expression level, is closer to natural collagen, and exhibits good biological activity.

The invention provides a trans-4-hydroxy-L-proline synthesis strain. The strain is bacillus cereus HBU-AI; the preservation unit is China General Microbiological Culture Collection Center (CGMCC) andthe preservation number is CGMCC No. 14164; the preservation date is May 17, 2017. The invention further provides an L-proline hydroxylase gene sequence of the strain and application of an L-proline hydroxylase gene to production of trans-4-hydroxy-L-proline. A result shows that the engineering strain containing the L-proline hydroxylase gene has a relatively high capability of synthesizing the trans-4-hydroxy-L-proline; when the strain is fermented for 52h, the concentration of hydroxyproline in a fermentation solution can reach 41.6g / L and the residual concentration of L-proline in a productis 0.8g / L.

The invention discloses a trans-4-hydroxy-L-proline hydroxylase modifying gene with high transformation rate and application thereof. An expression carrier of an optimization gene is constructed by optimizing nucleotide sequence of trans-4-hydroxy-L-proline hydroxylase (GenBankID:D78338.1) in Dactylosporangiumsp.RH1; the thallus is collected as the enzyme source of the transformation reaction through the prokaryotic expression. The experiment result expresses that the transformation rate of generating the trans-4-hydroxy-L-proline by using L-proline as a substrate is as high as 91%. The gene can be applied to biologically generating trans-4-hydroxy-L-hydroxyproline and has good industrial application value.

The invention relates to a DNA molecule, an expression vector, a recombinant bacterium and a construction method thereof, and a preparation method of hydroxyproline. The nucleotide sequence of the DNA molecule RBS1 is shown in SEQ ID NO.1. The RBS1 provided by the invention can improve the expression of proline hydroxylase coding gene.

The present invention provides a novel L-proline-4-hydroxylase, the amino acid sequence of which is shown in SEQ ID No.2, derived from Micromonospora sp. CNB394. The present invention also provides a genetically engineered bacterium E.coli HYP that efficiently expresses the above-mentioned L-proline-4-hydroxylase and synthesizes high-yield trans-4-hydroxyl-L-proline de novo. The genetically engineered bacterium has The advantages of good genetic stability and strong fermentative acid production ability, no plasmid, using the genome to express the above-mentioned novel L-proline-4-hydroxylase, using glucose and other cheap carbon sources as substrates to efficiently synthesize trans- 4-Hydroxy-L-Proline, the yield of trans-4-Hydroxy-L-Proline after 42 hours of fermentation is as high as 51g / L, which has a good industrial application prospect.

The invention discloses a gene segment provided with coded and highly-active trans-4-hydroxyl-L-prolyl hydroxylase and an application thereof. Gene codons of trans-4-hydroxyl-L-prolyl hydroxylase (GenBank ID: BAA 20094.1) in Dactylosporangiumsp. RH1 are optimized to obtain a p4hyd gene; the optimized p4hyd gene is truncated to obtain a P4Hyd1-257 gene; recombinant plasmids including pET-M-3C-P4Hyd and pET-M-3C-P4Hyd1-257 are built respectively, prokaryotic expression is performed through escherichia coli, and trans-4-hydroxyl-L-proline is produced in a conversion manner by utilizing thalli as an enzyme source. An experimental result shows that L-proline is taken as a substrate, compared with pET-M-3C-P4Hyd, the highest conversion rate of trans-4-hydroxyl-L-proline generated by pET-M-3C-P4Hyd1-257 in the conversion manner is increased from 91.4% to 97.4%, the conversion time is shortened from 80 hours to 60 hours, so that the conversion rate is improved remarkably, and the conversion time is shortened remarkably. The gene segment provided with coded and highly-active trans-4-hydroxyl-L-prolyl hydroxylase and the application thereof can be used for producing L-proline by a bioconversion method, and have better industrial application prospect.

The invention discloses a high-efficiency transformation method for producing trans-4-hydroxy-L-proline by a biotransformation method, which specifically includes a high-activity self-induced expression method of L-proline hydroxylase in recombinant E. The type, concentration and conversion conditions of the auxiliary agent in the high-efficiency conversion system of converting -proline into trans-4-hydroxy-L-proline, for the establishment of a biotransformation method for the industrial production of trans-4-hydroxy-L-proline Amino acid technology provides an important scientific basis.

The invention discloses the application of a prolyl hydroxylase inhibitor in the preparation of a drug for improving the symptoms of hepatolenticular degeneration disease. The prolyl hydroxylase inhibitor used in the invention can stabilize HIF‑1α and activate it. Transcriptional activity; the gray scale of the liver was weakened, and the expression of lipid metabolism factors was significantly reduced. H&E staining of the liver showed that prolyl hydroxylase inhibitors can effectively improve liver pathology and reduce liver copper accumulation; Expression of neuronal signaling and factors related to dopaminergic neuronal signaling was improved, and motor performance was improved.

The present invention pertains to the field of pharmaceutical chemicals, and relates to a substituted acethydrazide derivative, preparation method and use thereof; preferably, relates to a compound of Formula I or a pharmaceutically acceptable salt thereof. The compound of the present invention or a pharmaceutically acceptable salt thereof can effectively inhibit proline hydroxylase, stabilize HIF-α, particularly HIF-1α, and has potency in the manufacture of a medicament for treatment and / or prophylaxis and / or adjuvant therapy of anemia, acute ischemic reperfusion injury.

The invention discloses a method for producing trans-4-Hyp (trans-4-hydroxy-L-proline) through transformation of recombinant escherichia coli, and belongs to the technical field of biological engineering. A P4H enzyme derived from unulturated bacterium esnapd2 is cloned, related modification is performed on the basis of the P4H enzyme to obtain a mutant, and the recombinant Escherichia coli expressing the L-proline hydroxylase or the mutant thereof is constructed. The recombinant escherichia coli expressing the L-proline hydroxylase or the mutant of the L-proline hydroxylase is subjected to whole-cell transformation to produce Hyp, the yield of trans-4-Hyp after 24h of transformation is 5.38 g / L, and the transformation rate can reach 94.5%. A foundation is provided for industrial production of Hyp by microorganisms.