95 results about "Chemo enzymatic" patented technology

The invention belongs to the technical field of biology, and particularly relates to a compound for treating myocardial infarction. The chondroitin sulfate oligosaccharide is extracted from natural chondroitin sulfate, and after the chondroitin sulfate oligosaccharide is treated into an oligosaccharide chain through a chemical enzyme method, experiments prove that the chondroitin sulfate oligosaccharide is beneficial to the survival of myocardial cells. Compared with other products with similar functions, the glycopeptide polymer hydrogel formed after introduction of the polymer skeleton overcomes the problem of batch-to-batch difference of natural products and is clear in structure, and the hydrogel has excellent biological activity and does not need to entrap other active ingredients such as drugs. The effects of supporting the ventricular wall of the infarction region and promoting the survival of myocardial cells are obvious, the myocardial infarction area can be effectively limited, the thickness of the ventricular wall of the infarction region is increased, the damage of myocardial infarction to the heart is reduced, and the application prospect is very good.

The invention relates to heparinase III with high activity on 6-sulfated heparin and application of the heparinase III. The amino acid sequence of the heparinase III (BeHepIII) is as shown in SEQ ID NO. 2, and the nucleotide sequence of the coding gene is as shown in SEQ ID NO. 1. The invention discovers and discloses a novel heparinase III (BeHepIII) for the first time, the heparinase III is derived from Bacteroides eggerthii, has a higher recombinant expression level in escherichia coli, has very high degradation activity on heparin containing a 6-site sulfated structure, and particularly has the enzyme activity which is 93.5 times of that of commercialized heparinase FhHepIII at present, and the heparinase III can be used for degrading heparin containing 6-site sulfated structure. Therefore, the heparinase III (BeHepIII) provided by the invention has a wide application potential in heparin degradation. In addition, by using heparinase III (BeHepIII), directional excision of a reducing end of heparin oligosaccharide synthesized by a chemical enzyme method is realized, and complete chemical enzyme method synthesis of a pentasaccharide structure with anticoagulant factor Xa activity is successfully realized.

The invention discloses a nitrilase mutant and application of the nitrilase mutant in synthesis of a brivaracetam chiral intermediate, and provides an efficient (R)-3-cyanohexanoic acid nitrilase synthesis process by constructing the nitrilase mutant with improved catalytic activity and stereoselectivity. The requirement of industrially synthesizing the brivaracetam key chiral intermediate is met, and a foundation is laid for industrial production of brivaracetam synthesized by a chemical-enzyme method.

The invention discloses an olefin reductase, a coding gene, a carrier, an engineering bacterium and an application of the olefin reductase. An olefin reductase SeER mutant is obtained by carrying out single-point or multi-site site-directed mutagenesis or site-directed saturation mutagenesis on the 38th site, the 117th site, the 248th site, the 293th site and the 373th site of an amino acid sequence shown in SEQ ID NO.2. The invention provides a series of novel olefin reductase SeER mutants capable of efficiently, stereocomplementarily reducing an olefin compound to prepare a chiral compound, chiral GABA derivative drugs such as phenibuterol, baclofen, tolybuterol, pregabalin and the like are successfully synthesized through a chemical-enzyme coupling method, and compared with an existing method, the novel olefin reductase SeER mutants have the advantages that the novel olefin reductase SeER mutants can be used for efficiently and stereocomplementarily reducing the olefin compound to prepare the chiral compound; the strain disclosed by the invention can flexibly regulate and control the configuration of a product, and is high in stereoselectivity and conversion rate.

The invention discloses a chemical-enzyme integrated catalyst for oxygen functionalization reaction of an inert C-H bond. The chemical-enzyme integrated catalyst is obtained by simultaneously fixing a rhodium (III) complex and non-specific peroxygenase AaeUPO by a covalent organic framework (COF) material with a macroporous structure inside, wherein trialdehyde phloroglucinol and 5, 5 '-diamino-2, 2'-dipyridyl are used as ligands, polystyrene spheres are used as hard templates to prepare holes, and a covalent organic framework (COF) material with a macroporous structure is prepared, and is recorded as macro-COF. The chemical-enzyme integrated catalyst provided by the invention not only realizes efficient conversion of in-situ production of hydrogen peroxide to drive AaeUPO to catalyze an inert C-H bond oxygen functionalization reaction, but also can effectively inhibit damage of high-concentration hydrogen peroxide to an AaeUPO active center.

The invention relates to the field of biosynthesis high polymer materials, in particular to a biosynthesis method and application of omega-hydroxy fatty acid polyester (omega-PHFA) based on a whole-cell catalytic system. The method comprises the following steps: constructing a recombinant microbial whole-cell catalysis system containing P450 monooxygenase, CoA ligase and acyltransferase, and synthesizing omega-PHFA with a C12-C18 long carbon chain skeleton by taking renewable fatty acid as a substrate. Compared with a traditional chemical / enzymatic synthesis method, the technology is greener, and the performance defects of existing biosynthetic PHA materials are overcome. The invention provides related genetically engineered bacteria and a method for producing omega-PHFA by using the genetically engineered bacteria, and the genetically engineered bacteria have a wide application prospect.

The invention discloses a chemical-enzymatic coupling synthesis method of oseltamivir phosphate, which comprises the following steps: by taking monasic acid OS-01 as a substrate, sequentially carrying out S1 esterification, S2 ketalation, S3 selective ring opening, S4 TEMPO oxidation, S5 primary transaminase reaction, S6 acetylation, S7 secondary transaminase reaction and S8 phosphorylation reaction to obtain the final product oseltamivir phosphate OS-09. According to the method, the step of sodium azide required by a traditional route is avoided, the production safety and the industrialization possibility are enhanced, two different transaminases are used for respectively constructing different chiral centers, the steps of protecting groups and deprotecting groups on the traditional route are not needed, the overall route is short, the used chemical reagents are few, the yield is high, and the method is suitable for industrial production. The industrial requirements of environmental protection and low cost are met.

This invention relates to the fields of agricultural product processing and bio-feed technology, specifically to a method and application for detoxifying corn steep liquor. The method includes: subjecting a mixture of corn steep liquor, a composite color-protecting agent, and a composite catalyst to a first reaction; allowing the reaction system after the first reaction to stand in a mixture with a composite adsorbent; subjecting the mixture containing the reaction system after the stand-in reaction and a composite enzyme preparation to stand-in treatment, mixing treatment, and enzymatic hydrolysis; and subjecting the mixture containing the reaction system after the enzymatic hydrolysis and an activated composite microbial agent to anaerobic fermentation. This method employs a multi-stage chemical-enzyme-fermentation linkage process, achieving efficient detoxification of corn steep liquor while significantly improving flavor and nutrition. This method synergistically removes mycotoxins and sulfites, and enhances palatability and probiotic value through fermentation transformation of off-flavor substances and enrichment of small peptides and lactic acid. The process is simple and requires no adsorbent separation.

The invention relates to the technical field of preparation of electrochemical sensors, and particularly discloses a preparation method of a novel electrochemical enzyme sensor for profenofos pesticide detection based on an enzyme inhibition action principle, and the preparation method comprises the following steps: S1, treating a glassy carbon electrode to obtain a bare glassy carbon electrode GCE; s2, obtaining a composite material modified electrode Au50Ag2 (at) G / GCE; s3, an enzyme-chitosan composite modified electrode KbE-CS / Au50Ag2 (at) G / GCE is obtained; and S4, finally obtaining the profenofos electrochemical enzyme sensor NF / KbE-CS / Au50Ag2 (at) G / GCE based on the enzyme inhibition effect. According to the present invention, the white kidney bean esterase derived from plants is adopted as the detection enzyme source, and the electro-catalysis amplification effect of the Au50Ag2 (at) G composite material is adopted to construct the novel electrochemical enzyme sensor, and the method is the profenofos pesticide detection technology based on the enzyme inhibition effect principle;

The invention belongs to the technical field of biological medicine and disease prevention and treatment, and particularly relates to an SSEA4 conjugated virus-like particle anti-cancer vaccine as well as a preparation method and application thereof. Specifically, according to the vaccine, an SSEA4 antigen is efficiently synthesized through a chemical enzyme method, then the SSEA4 antigen is covalently coupled with a virus-like particle (VLP), the SSEA4 antigen and the VLP are combined with a monophosphoryl lipid A adjuvant with high safety, an organism can be induced to generate strong and lasting SSEA4-resistant humoral immune response, the generated antibody can specifically recognize and kill SSEA4 positive tumor cells, and the SSEA4 positive tumor cells can be effectively killed. Meanwhile, good safety is shown in an animal model, a new solution is provided for immunotherapy of SSEA4 positive tumors, and therefore the SSEA4 positive tumor immunotherapy polypeptide has wide application prospects and market value.

The invention belongs to the field of feed protein processing, and particularly relates to a method for improving the bioavailability of animal feed protein through chemical enzymolysis. The method comprises the following steps: mixing a compound protein raw material with water to form a suspension; sodium tripolyphosphate is added for phosphorylation pretreatment, so that protein is subjected to specific modification; after the pH is adjusted to be acidic, pepsase and acid phosphatase are added at the same time for an enzymolysis reaction; and after the reaction is finished, performing enzyme deactivation and drying to obtain the modified feed protein. According to the method, a specific enzyme cutting site is created on a protein molecule through chemical phosphorylation, then directional hydrolysis is achieved through efficient recognition of pepsin on the site, and meanwhile dynamic regulation and control of the hydrolysis process are achieved by means of the dephosphorylation effect of acid phosphatase. The obtained product has the outstanding advantages of high content of small peptides, good digestive absorbability, low bitterness, strong biological activity and the like, and the bioavailability of feed protein is remarkably improved.

The invention relates to a chemoenzymatic synthesis method of a high-glucose-type homogeneity mucoprotein core structural domain. The method comprises the following steps: firstly, catalytically synthesizing a key alpha-linked glycosylated amino acid block Fmoc-GalNacalpha-Ser / Thr-OH at a low temperature (-20 DEG C to 0 DEG C); then assembling the building blocks on a repeated polypeptide skeleton rich in proline, serine and threonine at accurate intervals (3-5 amino acid residues) through a solid-phase peptide synthesis technology to form a'sugar chain growth scaffold '; then, a series of high-specificity glycosyl transferases are used for carrying out sequential and directional sugar chain enzymatic extension on the scaffold, and a uniform core structure (such as a core type 3) is constructed; and finally, performing high-efficiency purification by a three-step series method of size exclusion chromatography, ion exchange chromatography and lectin affinity chromatography. The technical bottleneck of inhomogeneity (lt, 70%) of glycoforms in a traditional method is overcome, the mucoprotein core fragment with glycoform homogeneity larger than or equal to 95% and bioactivity highly similar to that of natural mucoprotein can be prepared on a large scale, and the method has wide application prospects in the biomedical fields of mucous membrane protective agents, drug delivery carriers and the like.

This invention discloses a class of pentacyclic triterpenoid derivatives with diverse B-ring modifications, their preparation methods, and applications. This invention utilizes a chemical enzymatic method to expand the chemical space of the B-ring in pentacyclic triterpenoids, obtaining a series of pentacyclic triterpenoid derivatives with diverse B-ring modifications. Pharmacological activity screening of these pentacyclic triterpenoid derivatives revealed their good tumor-suppressive activity, providing a promising application prospect for the prevention and treatment of tumors. This invention achieves the efficient synthesis of pentacyclic triterpenoid derivatives with diverse B-ring modifications, structures that are difficult to obtain using traditional chemical methods. Systematic antiproliferative activity screening identified candidate molecules with broad-spectrum and highly efficient inhibitory effects against various human tumor cell lines. These molecules show good drug development potential, providing novel lead compounds for the innovative development of anticancer drugs.

The application uses cheap and easily available hexose acid as a starting material, first obtains two precursors of 2,5-furan dicarboxylic acid synthesis, namely hexahydroxy acid and 4-deoxy-5-dehydro-hexahydroxy acid through enzyme catalytic reaction, and then 4-deoxy-5-dehydro-hexahydroxy acid is dehydrated under acidic conditions to obtain 2,5-furan dicarboxylic acid.

The invention discloses an electrochemical enzyme biosensor based on metal organic framework material ZIF8 and its application. The electrochemical enzyme biosensor uses a glassy carbon electrode as a working electrode, modifies a complex including GR, ZIF8 and AChE on the surface of the glassy carbon electrode, and encapsulates it with CS, and the structure is GCE / GR-ZIF8-AChE-CS. The electrochemical enzyme biosensor of the invention has a simple structure, uses the metal organic framework compound ZIF8 as a load material, has good biological inertness, a large load capacity, can be applied to the detection of isothiocarb, and has high detection sensitivity.

The application discloses a cytochrome P450BM3 mutant, a carrier, microorganisms and application thereof, and belongs to the technical field of chemical enzyme synthesis.The application discloses a synthetic route of a key drug intermediate dehydro-nandrolone acetate synthesized by a chemical enzyme method, wherein enzyme catalysis and chemical catalysis are included.The enzyme catalysis includes a cytochrome P450 mutant and 17beta-steroid reductase, and a one-pot two-step method is used to catalyze acid to be removed to generate 7beta-hydroxynandrolone.Combined with chemical catalysis, the 7beta-hydroxynandrolone is subjected to dehydration esterification to generate the dehydro-nandrolone acetate.The synthetic route has short steps, high yield and the ability to reduce the production cost of related steroid drugs.

Described herein are 4-N-derivatized sialic acid compounds and efficient chemoenzymatic methods for synthesizing sialosides containing the 4-N-derivatized sialic acid. Methods of using the 4-N-acyl sialic acids and sialosides are also described herein. Described also herein are 8-N- derivatized sialic acid compounds and methods for synthesizing and using the same.

Presented are kainoids, kainoid precursors, and kainoid derivatives and synthetic schemes for producing kainoids, kainoid precursors, and kainoid derivatives. The synthetic schemes for producing kainoids can make use of kainoid synthases, for example, in cyclizing a kainoid precursor to produce a kainoid. The kainoids can include an alkyne group 5 to facilitate the production of kainoid derivatives.

Disclosed is a method for forming selenoneine or analogs thereof. The method may include phosphorylating sodium selenide to a selenophosphate, using adenosine triphosphate (ATP) and at least a first protein, generating a selenosugar by converting the selenophosphate using at least a second protein in the presence of a common sugar donor, and forming selenoneine or an analog thereof by combining the selenosugar with N,N,N-trimethyl-L- histidine or analogs thereof using at least a third protein. The method may include combining SenA, SenB, and SenC in an aqueous buffer at neutral pH and ambient temperature, and allowing SenA, SenB, and SenC to form selenoneine or an analog thereof in the presence of ATP, a common sugar donor, and sodium selenide.

The present invention discloses a method for mechanochemical enzyme-coupled lignin degradation. The specific operation process is to place lignin powder, a solid-phase reagent, and zirconium oxide beads in a polytetrafluoroethylene ball mill, perform a co-grinding reaction in the ball mill, add an enzyme after ball milling, add a solvent for stirring, and reflux reaction. After the reaction, the mixture is dissolved in water and extracted with ethyl acetate. The insoluble residue is centrifuged, washed with ethyl acetate, and vacuum dried for detection and analysis. The present invention has the advantages of a short process route, low reagent usage, simple operation, low cost, low pollution, low molecular weight of the obtained product, and good degradation efficiency. It is a lignin degradation method with good promotion and application prospects.

The invention belongs to the crossing field of a biosensing technology and efficient screening of natural medicines, and particularly relates to an electrochemical enzyme biosensor based on synergistic interaction of a nano material and application of the electrochemical enzyme biosensor in rapid screening of anti-gout active components. In order to solve the problems of low efficiency, high cost, dependence on in-vitro biochemical detection, high false positive rate and the like in the traditional drug screening technology, high-throughput and high-sensitivity detection of anti-gout active components in natural products is realized by innovatively constructing a xanthine oxidase (XO) immobilized nano-composite sensing interface.

The invention provides an electrochemical microplate reader, which comprises: an acquisition module, which is used for acquiring an electric signal of a to-be-detected object; the interface module comprises an electrode interface unit and a communication interface unit, the electrode interface unit is used for forming a detection circuit of the acquisition module together with a detection electrode, and the communication interface unit is used for establishing control signal transmission and data transmission between the detection analysis module and the outside; the detection analysis module is used for performing quantitative analysis on the to-be-detected object according to the electric signal and optimizing a quantitative analysis result according to a machine learning algorithm; and the display module is used for displaying the result of the quantitative analysis.

The present invention provides a chemoenzymatic synthesis method for blood group trisaccharide, which includes: dissolving a substrate, GDP-Fuc, and magnesium chloride in water, adjusting the pH of the solution, adding the enzyme α1,2-FucT, and stirring for reaction to obtain a first product reaction solution; adding a de-enzyme solvent, standing, centrifuging and filtering, taking the supernatant, and evaporating to remove the de-enzyme solvent to obtain an aqueous solution of the first product; adding a glycosylated product, adjusting the pH after dissolution, and adding a glycosyltransferase to obtain a second product reaction solution; using a de-enzyme solvent to remove the glycosyltransferase, standing, centrifuging and filtering, taking the supernatant, and evaporating to remove the de-enzyme solvent to obtain an aqueous solution of the second product; separating and purifying the aqueous solution of the second product, and freeze-drying to obtain a solid product; dissolving the solid product, adding Pd / C, introducing hydrogen gas, stirring and filtering to remove Pd / C, concentrating and freeze-drying to obtain the blood group trisaccharide. This method provides a new technical idea for the chemoenzymatic synthesis of blood group trisaccharide, which is conducive to popularization and application and solves the problems of low yield and high price.

The invention discloses a method for degrading kitchen waste protein through mechanochemical enzyme coupling, which comprises the following steps: mixing a kitchen waste stock solution with a solid-phase reagent, putting the mixture into a polytetrafluoroethylene ball mill, adding stainless steel beads, setting the rotating speed of the ball mill to be 100-500rpm, carrying out a grinding reaction for 6-12 cycles, and filtering to remove residues after the ball-milling reaction is finished, the kitchen waste protein degradation liquid is obtained. The method has the advantages of short process route, less reagent dosage, simplicity in operation, low cost, less pollution, high protein degradation rate, large amount of obtained amino acid, good degradation efficiency and the like, can be used as a fertilizer to better promote plant growth, and is a technology with relatively good popularization and application prospects.

The invention firstly discloses a method for synthesizing the spraadine A and the 3-OH spraadine A by utilizing a chemical-enzyme method. The synthetic route mainly comprises three steps: 1) synthesizing (-)-beta-CPA through a novel chemical enzyme method synthetic route; (2) catalyzing the (-)-beta-CPA in vitro by using oxidation cycloaddition enzyme CpaO to carry out intramolecular asymmetric oxidation [3 + 2] cycloaddition reaction to generate (-)-alpha-CPA; and (3) by taking the (-)-alpha-CPA as a substrate, synthesizing alpha-CPA natural products in a high oxidation state, such as speradine A, 3-OH speradine A and the like, through chemical modification. The invention has important theoretical and practical significance for three-dimensionally and specifically preparing or synthesizing a series of high-oxidation-state alpha-CPA natural products, and provides a leading compound for the development of anti-cancer drugs.

Provided herein are synthetic methods and intermediates useful in the preparation of polyketides, including 12-membered ring macrolides. These synthetic methods and intermediates provide scalable access to polyketides, including polyketides with potent bioactive splicing modulator activity and therapeutic potential, including treating various types of cancer. The polyketides include 12-membered ring macrolides, including Compound A, Compound B, and pladienolides, including pladienolide A, pladienolide B, pladienolide C, pladienolide D, pladienolide E, pladienolide F, and pladienolide G.

The invention discloses a method for extracting ganoderma lucidum polysaccharide by a microwave-assisted green chemical-enzyme method and application, which comprises the following steps: taking ganoderma lucidum sporocarp powder, adding cellulase and water, uniformly mixing, then adding 30% hydrogen peroxide, uniformly mixing, adjusting the pH value to 3-7, and carrying out microwave-assisted extraction for 40-120 minutes under the conditions that the power is 400-800W and the temperature is 50-90 DEG C; and centrifuging, taking supernate, concentrating to 30-50% of the original volume, and freeze-drying to obtain the ganoderma lucidum polysaccharide. The method is high in ganoderma lucidum polysaccharide extraction efficiency and more complete in ganoderma lucidum polysaccharide activity retention. The ganoderma lucidum sporocarp polysaccharide product is free of peculiar smell and good in sensory property, the SOD activity in C.elegans under the hydrogen peroxide stress state can be improved by 33.76%, the CAT activity can be improved by 229.36%, and the MDA content can be reduced by 85.65%.

A field of glycoproteomics for identifying novel cell surface O-linked glycopeptide epitopes. The disclosure relates to methods for identifying O-linked glycopeptides, more specifically, Tn- or SiaTn-antigen O-glycosylation sites on cell surface proteins, the method comprising a combined workflow of enzymatic treatments of cells or tissue samples to allow specific enrichment and labelling of Tn antigens, followed by mapping of the O-glycopeptides through LC-MS / MS. More specifically, the disclosure relates to a chemo-enzymatic method to produce labelled O-glycan-peptides from a plasmamembrane-protein extract sample, wherein Sialyltransferase (ST3Gal1 / ST3Gal3) treatment is used to protect free galactose against a follow-on treatment with Galactose oxidase, wherein the oxidized glycans are simultaneously labelled. The disclosure further relates to a tandem mass spectrometry method using an enriched TMT-labelled (Sia)Tn-antigen glycopeptides in a dual fragmentation triggered approach to resolve the identity, localization and quantification of the O-linked glycan peptides.

The invention discloses a method for screening a DNA antibody by using a solid-phase chemical enzyme, which comprises the following steps: collecting a body fluid sample, and extracting a DNA sample in the body fluid sample; and capturing DNA combined with the antibody or capturing an anti-DNA antibody. DNA methylation is a crucial epigenetic modification, and is obviously changed in various diseases. These changes may result in gene expression imbalance, resulting in the development and progression of diseases. Diseases associated with altered DNA methylation include cancer, neurodegenerative diseases, autism, autoimmune diseases, metabolic disorders, and developmental disorders. Selection of specific antibodies for altered DNA methylation patterns can be a valuable tool for disease diagnosis and treatment. By aiming at these specific modifications, researchers and clinicians can develop more accurate diagnostic tests, determine therapeutic objectives, and monitor disease progression and therapeutic response.

The present invention discloses a method for synthesizing danshensu using a chemoenzymatic cascade system. The first enzymatic cascade system or the second enzymatic cascade system is subjected to a catalytic reaction to generate (R)-2-hydroxy-3-(4-hydroxy-3-methoxyphenyl) propionic acid, and then a demethylation reaction is carried out using hydrobromic acid to finally generate D-danshensu. The technical solution provided by the present invention synthesizes danshensu using vanillin as a raw material. Vanillin has a low cost and stable chemical properties; the reaction conversion rate of the chemoenzymatic cascade system for synthesizing danshensu is high, the conversion rate of vanillin can reach 100%, and it has excellent chemoselectivity. The yield of danshensu reaches 90%, all of which are D-danshensu, and no other by-products are generated, showing good application prospects.