53 results about "Chemo enzymatic" patented technology

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 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 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.

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 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 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.

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 an olefin reductase SeER mutant and application thereof in synthesis of chiral drugs, and provides a series of novel olefin reductase SeER mutants capable of efficiently and asymmetrically reducing beta-alkyl / aryl-beta-cyanoacrylate compounds, and the olefin reductase SeER mutants have high conversion rate (gt; 99% conversion rate), good enantioselectivity (gt; 99% ee), the substrate range is wide (gt; according to the present invention, the screened olefin reductase SeER mutant can be successfully synthesized into the chiral GABA derivative drugs such as pregabalin and brivaracetam through the chemical-enzyme coupling method, such that the overall reaction yield is improved, and the route is simple;

The invention discloses a method for synthesizing ergothioneine based on a chemical-enzyme method, and belongs to the technical field of biology. The method comprises the following steps: firstly, synthesizing histidine betaine by using a chemical method, and carrying out selective trimethylation reaction in an alkaline aqueous solution by using methyl iodide as a methyl donor, so that the final concentration of histidine betaine is 77g / L, and the conversion rate is 93%; then, histidine betaine sulphide enzyme EanB from an anaerobic biological argillobiosis source is selected, a histidine betaine sulphide enzyme mutant F97A / T387M / E277K / N354Q / F441S / S335K is constructed through truncation transformation and mutation, and histidine betaine is directly vulcanized to synthesize ergothioneine. According to the present invention, through the cascade reaction of the chemical-enzyme method, the synthesis from the histidine to the ergothioneine is successfully achieved, the final ergothioneine yield is 33.5 g / L, and the conversion rate is 95%;

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

The application discloses an olefin reductase, a coding gene, a carrier, an engineering bacterium and application thereof, wherein the olefin reductase SeER mutant is obtained by single-point or multi-point site-directed mutation or site-directed saturation mutation of the 38th, 117th, 248th, 293rd and 373rd sites of the amino acid sequence shown in SEQ ID NO. 2. The application provides a series of new olefin reductase SeER mutants capable of efficiently stereocomplementary reduction of olefin compounds to prepare chiral compounds, and successfully synthesizes chiral GABA derivative drugs such as phenibut, baclofen, tolperisone and pregabalin through a chemical-enzyme coupling method. Compared with existing methods, the strain can flexibly regulate the configuration of the product, and has high stereoselectivity and conversion rate.

The invention relates to uridine diphosphate-N-difluoroacetyl galactosamine as well as a preparation method and application thereof. The chemical structure of the UDP-GalNDFA is similar to that of a natural substrate UDP-GalNAc, and the core difference is that N-acetyl at the site 2 of the UDP-GalNDFA is replaced by N-difluoroacetyl. The preparation method comprises the following steps: (1) preparing difluoroacetyl galactosamine; and (2) preparing the UDP-GalNDFA by taking the difluoroacetyl galactosamine as a substrate. As a novel artificial donor sugar, the UDP-GalNDFA has the characteristics of high activity, difficulty in hydrolysis and capability of being artificially synthesized. Compared with the existing donor sugar UDP-GalNTFA, the glucose UDP-GalNTFA has higher catalytic activity on glycosyl transferase, has stronger stability, is not easy to hydrolyze, and can be used as a donor substrate for synthesizing a chondroitin oligosaccharide skeleton, a chondroitin oligosaccharide intermediate and a chondroitin oligosaccharide derivative by a chemical enzyme method.

The present application relates to a kind of high activity, high thermal stability heparin N-sulfate group transferase mutant and its coding gene and application.The amino acid sequence of the N-sulfate group transferase mutant M8 is as shown in SEQ ID NO.2, and the nucleotide sequence of coding gene is as shown in SEQ ID NO.1.The N-sulfate group transferase mutant M8 is 24 amino acids in wild-type N-sulfate group transferase Mutations.Determination of stability, compared with wild-type N-sulfate group transferase at 37 DEG C only incubate less than 1 day, all activity is lost, the N-sulfate group transferase mutant M8 of the present application has high activity and extremely strong high thermal stability, can be incubated at 37 DEG C 7 days still keep about 50% activity.The present application improves the efficiency of heparin chemical enzyme synthesis, expands the application range of N-sulfate group transferase mutant, makes it possible to synthesize heparin at high temperature, greatly promotes the application development of heparin biomimetic synthesis.

Described herein are various methods of removing uracils due to the deamination of unmethylated cytosines in an assay using engineered cytosine deaminases to deaminate methylated cytosines. The Chemoenzymatic Uracil Replacement of Nucleobases (ChURN) method includes providing a sample comprising single stranded DNA library fragments in which a cytosine deaminase has deaminated methylated cytosines; contacting the sample with an uracil DNA glycosylase (UDG) to deglycosylate uracil residues to form abasic sites having a hemiacetal formation within the single stranded DNA library fragments; contacting the sample with a reactive cytosine nucleobase analog to install a cytosine at abasic sites thru a noncanonical linkage; and subjecting the sample to polymerase chain reaction (PCR) amplification resulting in double stranded DNA corrected library fragments. The Uracil Enzymatic Removal and Substitution at Errors (U-ERASE) method includes providing a sample comprising single stranded DNA library fragments in which a cytosine deaminase has deaminated methylated cytosines, synthesizing double stranded DNA library fragments in which the second strand is tagged to facilitate its downstream degradation, treating the double stranded DNA library fragments with an uracil DNA glycosylase and an endonuclease resulting in the removal of uracil bases and single nucleotide gaps at those sites, and repairing the single nucleotide gaps through treatment with a polymerase, dCTP, and a ligase, resulting in replacement of false positive uracil bases with a mismatched base, such as cytosine. Subsequently, the second strand is selectively degraded, allowing for selective amplification of the original DNA strand via PCR.

The invention discloses a chemical enzyme synthesis method of chiral aporphine alkaloids, and relates to the field of biochemistry, the method comprises the following steps: taking a phenylethylamine compound and a phenylacetic acid compound as initial raw materials, and carrying out amide condensation and dehydration cyclization to form a compound IV; then reducing through imine reductase or an imine reductase mutant to form a V-type compound; then, a compound VI is formed through a methylation reaction of methyltransferase or Eschweiler-Clarke or methyl iodide, and the compound VI is formed through a methylation reaction of methyltransferase or Eschweiler-Clarke or methyl iodide; and finally, carrying out photocatalytic coupling to obtain the aporphine alkaloid. According to the preparation method disclosed by the invention, purification is not needed in the preparation reaction process from the IV-type compound to the VI-type compound, and the compound can be directly used for the last step of photocatalytic coupling; the optically pure (S)-VII aporphine alkaloid can be obtained; the method is mild in reaction condition, green, environment-friendly, simple in process, low in cost, capable of reaching gram-level scale and suitable for industrial production.

The invention discloses a method for synthesizing quinoxaline and derivatives thereof by using a chemical enzyme concerted catalysis system, which is characterized in that 2-(2-amino anilino) ethanol compounds are used as substrates, and the quinoxaline and the derivatives thereof are synthesized in a solvent through chemical enzyme concerted catalysis; wherein the chemical enzymes comprise alcohol dehydrogenase, catalase, natural cofactors and artificial cofactors. The quinoxaline compound disclosed by the invention is one of quinoxaline, 6-fluoroquinoxaline, 6-chloroquinoxaline, 6-bromoquinoxaline, 6-methylquinoxaline, 6-methoxyquinoxaline, 6-hydroxyquinoxaline and the like. The formula of the quinoxaline compound is shown in the description. According to the technical scheme provided by the invention, addition of additional cosubstrates and coenzymes is avoided, the highest substrate conversion rate can reach 95%, and excellent chemical selectivity is achieved. The product synthesized by the synthesis method is good in selectivity and relatively high in yield, and has a relatively good industrial application prospect in preparation of quinoxaline compounds through biological catalysis.

A method of preparing glucodiamine, comprising contacting glucose with an enzyme oxidation catalyst under conditions suitable for the formation of glucodialdose; contacting glucodialdose with a nitrogen-containing compound under conditions suitable for the formation of glucodioxime; and reducing glucodioxime under conditions suitable for the formation of glucodiamine. A method of preparing glucodiamine, comprising contacting glucodioxime with a dehydration catalyst under conditions suitable for the formation of glucodinitrile; and reducing glucodinitrile under conditions suitable for the formation of glucodiamine. A chemoenzymatic method of producing a bio-based amide platform chemical, comprising: contacting glucose with a biocatalyst under conditions suitable for the formation of glucodialdose; contacting glucodialdose with a base under conditions suitable for the formation of glucodioxime; and contacting glucodioxime with a hydrogenation catalyst in the presence of hydrogen under conditions suitable for formation of glucodiamine.

The invention provides a controllable acetyl removal method of heparin precursor polysaccharide, which comprises the following steps: (1) mixing heparin precursor polysaccharide from microbial fermentation with an alkaline aqueous solution, and reacting at 40-70 DEG C for 1-4 hours to obtain a reaction solution A; wherein the concentration of the alkaline aqueous solution is 1-4M; the mass ratio of the heparin precursor polysaccharide to the solute in the alkaline aqueous solution is 1: (1-4); and (2) adding an acidic regulator into the reaction liquid A to regulate the pH value to 6.5-7.5, and then carrying out ethanol precipitation and vacuum drying treatment to obtain the heparan precursor polysaccharide. The method has the beneficial effects that the method has the advantages of high efficiency, stability, high reaction efficiency, controllable conditions, good batch repeatability, low cost, simple process, easiness in amplification and the like, and is very suitable for industrial large-scale production; according to the method, 80%-95% of deheparan precursor polysaccharide meeting subsequent enzymatic sulfonation requirements can be stably obtained, and the chemical enzymatic synthesis process of non-animal source heparin is effectively promoted.

A chemoenzymatic method for synthesizing steroid HE3286 is provided, comprising: screening cytochrome P450 mutant LG-23 capable of catalyzing 7β-hydroxylation of dehydroepiandrosterone; enzymatically converting dehydroepiandrosterone to 7β-hydroxy-dehydroepiandrosterone using the P450 BM3 mutant enzyme; and chemically performing alkynylation at the C17th position carbonyl group to generate steroid HE3286. The steroid HE3286 synthesis method not only features simplified synthetic steps and high catalytic selectivity, but also offers mild reaction conditions, low cost, and environmentally friendly efficiency. This approach holds significant application value for advancing the development of steroid pharmaceuticals.

A chemoenzymatic manufacturing process for the preparation of high value chemicals includes the enzymatic oxidation of ethanol using an oxidizing biocatalyst to form an acetaldehyde intermediate. In addition, the process includes contacting of the acetaldehyde intermediate with a carboligating biocatalyst to form pyruvic acid. Further, the process includes reacting pyruvic acid with ethanol to form ethyl pyruvate. Still further, the process includes contacting the ethyl pyruvate with a metal catalyst and hydrogen to form ethyl lactate. The process also includes hydrolyzing the ethyl lactate to produce lactic acid. Moreover, the process includes contacting at least a portion of the lactic acid with a dehydrating catalyst to form acrylic acid. In addition, the process includes contacting at least a portion of the lactic acid with a metal catalyst and hydrogen to form propylene glycol and n-propanol.

The present disclosure relates to engineered bacterial glycoside-3-oxidases with improved catalytic and stability properties (and homologues) and their use in producing rare C3-sugar epimers. The present disclosure further relates to a method for the enzymatic oxidation of the C3-position of chemically protected saccharides at the C1-position, followed by a chemical reduction and deprotection steps.

The application discloses an O-GlcNAc modified peptide segment enrichment method and analysis method based on chemical enzymatic labeling and reversible oxime bond formation. The method comprises the following steps: removing N-glycan chains of a peptide segment mixture by using a glycosidase to obtain an N-glycan chain removed peptide segment mixture; mixing the N-glycan chain removed peptide segment mixture, UDP-GalNLeV and Gal-T1-Y289L mutant to perform an enzymatic labeling reaction to obtain a mixture containing ketone labeled O-GlcNAc peptide segments; covalently capturing the ketone labeled O-GlcNAc peptide segment mixture by using a solid phase carrier of allyl hydroxylamine, washing to obtain a solid phase carrier loaded with peptide segments; reacting the solid phase carrier loaded with peptide segments with a reagent containing a hydroxylamine group, collecting a release liquid, desalting to obtain O-GlcNAc peptide segments labeled by a methoxy oxime bond. The application realizes one-step labeling and reversible enrichment of O-GlcNAc modified peptide segments, and improves the coverage of glycopeptide identification.

The present invention relates to a method for preparing linear polylysine or polyarginine by chemoenzymatic bulk polymerization of lysine esters or arginine esters, respectively. The invention also relates to a method for controlling the regioselectivity of polylysine or polyarginine in the chemoenzymatic bulk polymerization of lysine esters or arginine esters.