96 results about "Enzyme complex" patented technology

Described herein are programmable DNA proteolytic target chimera complexes that can be used both for the direct treatment of cancer by inhibiting biochemical pathways that are overexpressed in cancer cells, and for the indirect treatment of cancer by recruiting the E3 ligase complex to engage with a protein of interest or a mutant thereof and initiating proteolysis. Also described herein are methods of using the complexes in the treatment of cancer, as well as compositions comprising the complexes.

The invention discloses a rice large-scale metabolism regulator DGP1 and crop metabolism quality design. The invention finds that the structural domain of the rice DGP1 family protein TIGR01589 targets a plurality of metabolic enzymes and promotes the metabolic enzymes to be transferred to a cell nucleus; the structural domain also has an intervention effect on a light breathing pathway by changing the composition of a glycine decarboxylase compound in mitochondria. Based on a molecular mechanism of DGP1, a chimeric protein ST containing a conservative structural domain and coupled with a chloroplast positioning signal is designed, and genetic engineering modification of pakchoi is realized. The modification strategy not only can guide substrate flow of a central glycolysis pathway to flavonoid biosynthesis, but also can promote accumulation of saccharides derived from photosynthetic products in leaves.

The present disclosure relates to novel CRISPR enzymes, reaction mixtures, methods of using the same in the detection of microorganisms, such as FluA, FluB, amongst others. The disclosure particularly relates to the discovery of novel functional CRISPR enzyme complexes, particularly systems displaying trans cleavage activity.

The invention belongs to the technical field of bioengineering, and relates to a construction method and application of recombinant yarrowia lipolytica for synthesizing plant antitoxin Kauralexin A1. The method is realized by constructing recombinant yarrowia lipolytica. The construction method of the recombinant yarrowia lipolytica for synthesizing the Kauralexin A1 comprises the following steps: overexpressing a codon optimized antitoxin Kauralexin A1 biosynthetic pathway in yarrowia lipolytica, screening adaptive P450 reductase to improve the electron transfer efficiency in a P450 system, and constructing a metabolic pathway scaffold-free multi-enzyme complex to enhance cascade biological catalysis and metabolic flux, so as to obtain the recombinant yarrowia lipolytica for synthesizing the Kauralexin A1. The expression of the mevalonic acid pathway is enhanced, so that a sufficient GGPP precursor is provided for the biosynthesis of the Kauralexin A1. The construction method of the yarrowia lipolytica for biosynthesizing the antitoxin Kauralexin A1, provided by the invention, is simple to operate, and the constructed yarrowia lipolytica can be used for efficiently producing the Kauralexin A1 and has relatively high production and application values.

The invention discloses a multi-enzyme compound noodle stretching agent and application thereof, and belongs to the technical field of food additives. The multi-enzyme compound noodle stretching agent is prepared from the following components in parts by mass: 1 to 10 parts of papain, 1 to 10 parts of trypsin, 1 to 10 parts of glucose oxidase, 1 to 10 parts of lipase and 10 to 30 parts of tea polyphenol. Through the synergistic effect of multienzyme compounding and tea polyphenol, the dough structure and the noodle processing performance are improved, the protein depolymerization and crosslinking degree and the protein oxidation degree of the dough can be regulated and controlled at the same time, the dough strength and ductility are considered, the cooking loss is reduced, meanwhile, the green and natural characteristics are kept, no chemical modifier is added, and the production cost is reduced. And the limitation of the existing single enzyme or chemical modifier is overcome, so that the method has a relatively good application prospect in the technical field of food additives.

The invention belongs to the technical field of synthetic biology and enzyme engineering, and particularly relates to a multi-enzyme in-vivo space co-localization carbon metabolic flux strengthening method, a genetically engineered bacterium and application of the genetically engineered bacterium. According to the invention, a recombinant expression vector is constructed, key enzymes RHLA and RHLB participating in synthesis of a target product are respectively subjected to fusion expression with protein scaffold modules RIAD and RIDD through (G4S) 3 flexible connecting peptide, and are introduced into host chassis bacteria, so that a stable multi-enzyme composite system is formed in cells, the transmission distance of a metabolic intermediate is shortened, and the expression efficiency is improved. Spatial co-localization and concerted catalysis of key catalytic enzymes are realized, and the conversion efficiency of a carbon source to a target product is improved. Compared with the prior art, the genetically engineered bacterium constructed by the invention has the advantages of stable structure, no need of complex regulation, reasonable metabolic flow distribution and the like in the fermentation process, the synthesis efficiency and yield of the target biosurfactant are remarkably improved, and the feasibility of industrial amplification application is enhanced.

This invention discloses a lipophilic yeast strain for synthesizing astaxanthin (Yersinia lipophila). Yarrowia lipolytica This invention relates to an engineered strain, its construction method, and its application. The engineered strain is a *Yersinia lipolytica* strain carrying CrtZ and CrtW. The invention utilizes genetic engineering methods to introduce a *Yersinia lipolytica* strain derived from *Rhodochophora* into a β-carotene-producing strain via genetic engineering. Haematococcus Pluvialis The β-carotene hydroxylase encoding gene (HpCrtZ) originates from the genus Paracoccus ( Paracoccus sp. The gene encoding β-carotene ketolase (PsCrtW) and the gene derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae) Saccharomyces cerevisiae The method involves obtaining a genetically engineered *Yersinia lipolytica* strain that produces astaxanthin by constructing a fusion enzyme complex and fusing it with peroxisomes, endoplasmic reticulum, and lipid droplet-targeting signal peptides to obtain a *Yersinia lipolytica* strain with higher astaxanthin yield. The astaxanthin-producing genetically engineered strain constructed in this invention achieved an astaxanthin yield of 1.43 g / L in a 5 L bioreactor after pH control and continuous feeding optimization.

The invention discloses a multi-enzyme complex of S-adenosylmethionine decarboxylase / spermine synthetase and an application of the multi-enzyme complex in production of spermine. Belongs to the technical field of enzyme engineering. The purpose of the present invention is to increase the yield of spermine. The invention provides a multi-enzyme complex of S-adenosylmethionine decarboxylase / spermine synthetase. The multi-enzyme complex is obtained by connecting the S-adenosylmethionine decarboxylase and the spermine synthetase through a flexible linker. And a theoretical basis is provided for preparation of spermine.

The invention provides a controllable photo-thermal-enzymatic degradation porous material as well as a preparation method and application thereof. The controllable photo-thermal-enzymatic degradation porous material is prepared from the following raw materials: an oil phase mixture and a water phase mixture, the oil phase mixture comprises an enzyme sensitive polymer base material, a random heteropolymer (RHP)-enzyme compound, Janus photo-thermal nanoparticles, an internal phase dispersing agent and a solvent; the aqueous phase mixture comprises water, electrolyte and alginate. According to the porous material provided by the invention, the Janus photo-thermal nanoparticles and the RHP-enzyme compound are coupled, under near-infrared light, RHP conformational change can be triggered to release BC lipase, degradation of the porous material can be triggered under the mild condition of 37 DEG C, and the degradation rate within 48 hours can reach 98%.

This application discloses a multi-enzyme complex based on a Snap-Fit self-assembly tag, its construction method, and its applications, relating to the field of protein engineering technology. The multi-enzyme complex based on the Snap-Fit self-assembly tag comprises: a TLL-FitC fusion protein with the amino acid sequence shown in SEQ ID NO.13, a FitN-AnChOx fusion protein with the amino acid sequence shown in SEQ ID NO.14, and an OleT protein with the amino acid sequence shown in SEQ ID NO.15. PRN - SnapG fusion protein. This multi-enzyme complex employs a Snap-Fit assembly strategy, which allows three different enzymes to be assembled using only one set of tags. When applied to the synthesis of olefins, it helps to improve the transfer efficiency of 1-olefin cascade reaction intermediates between upstream and downstream enzyme active sites, thereby increasing the final yield, and has great application potential.

The invention discloses a multistage enzyme and quorum sensing signal molecule MABR membrane and a preparation method thereof, through a bionic induction strategy, while the anti-pollution characteristic of the MABR membrane is maintained, multistage enzyme and quorum sensing signal molecules are fixed on the surface of the membrane, and the multistage enzyme and quorum sensing signal molecules induce microorganisms to secrete an extracellular polymeric substance (EPS), so that the binding strength of a biological membrane is improved by 40% or more. The preparation method of the multi-stage enzyme and quorum sensing signal molecule MABR membrane comprises the following steps: (1) screening nitrifying bacteria and extracting multi-stage enzyme and quorum sensing signal molecules of the nitrifying bacteria; (2) carrying out composite pretreatment on the surface of the MABR membrane, wherein the composite pretreatment comprises hydrophilic modification, functional group introduction, swelling of the surface of membrane silk and the like; and (3) immobilizing multistage enzyme and quorum sensing signal molecules (including cell membrane fragments, periplasmic space enzyme complexes and the like) in the strain by adopting an immobilization technology to form a bioactive interface layer with the thickness of about 50-200nm.

The invention provides a beta-galactosidase compound and a method. The beta-galactosidase compound comprises beta-galactosidase and cyclic adenosine monophosphate combined with amino acid residues of the beta-galactosidase. According to the invention, small molecular cyclic adenosine monophosphate acts on lactase molecules, and the catalytic activity of lactase is significantly enhanced through unique intermolecular interaction. Compared with a traditional lifting means, the method has the advantages that the lifting efficiency is higher, an ideal lactase activity level can be achieved in a shorter time, and a powerful guarantee is provided for lactose digestion and utilization.

The invention provides a CBM mutant, a cellulose microsphere immobilized multienzyme compound and application, belongs to the technical field of immobilized enzymes, and particularly provides a CBM mutant CBM-4, the amino acid sequence of which is as shown in SEQ ID NO.1; the invention relates to a carrier compound for immobilizing an enzyme, which comprises cellulose microspheres and one of a fusion protein ScafCCR-CBM-4 or a fusion protein ScafCCR-CBM-1, the invention discloses a preparation method of cellulose microspheres and a cellulose microsphere immobilized multienzyme compound. According to the cellulose microsphere immobilized multienzyme compound prepared by the method, the conversion rate of trehalose is increased, multiple times of cyclic utilization can be achieved, the utilization rate of enzyme is greatly increased, and the production cost is reduced.

The application discloses a method for constructing a multi-enzyme complex engineering strain, and has the characteristics that the method comprises the following steps: S1, constructing a light decarboxylase FAP and an oil-producing yeast fatty acid synthase fusion expression vector; S2, electrically shocking and transforming the constructed target vector into an agrobacterium competent AGL1, and introducing the target vector into a starting strain through an agrobacterium-mediated transformation method to obtain an engineering strain that is back-supplemented with a fatty acid defect; the engineering strain that is back-supplemented with the fatty acid defect is obtained, the FAP-FAS2 fusion expression is realized at the same time as the self-assembly of the intracellular FAS1, and a FAP-FAS synthase complex is obtained. The complex not only back-supplements the fatty acid defect function, so that the strain grows normally, but also can synthesize alkanes in the process of blue light irradiation fermentation culture, and realizes the intracellular assembly of protein macromolecules.

The invention relates to a rhodium nano-enzyme compound and application thereof in preparation of drugs for preventing, relieving and / or treating drug-induced liver injury. The rhodium nano-enzyme compound comprises an extracellular vesicle simulant and rhodium nano-enzyme located in the extracellular vesicle simulant. The surface of the rhodium nano-enzyme is coated with the extracellular vesicle simulant, the extracellular vesicle simulant has good biocompatibility and low immunogenicity and can promote tissue repair and regeneration, and the extracellular vesicle simulant can be removed in a non-specific mode and then is enriched in the liver in a targeted mode, so that the rhodium nano-enzyme is enriched in the liver together with the extracellular vesicle simulant. The rhodium nano-enzyme has excellent simulation activity of superoxide dismutase (SOD) and catalase (CAT), can efficiently remove various active oxygen species, and can inhibit inflammatory reaction caused by APAP, so that the rhodium nano-enzyme has a targeted treatment effect on drug-induced liver injury.

The invention relates to the field of bio-enzyme catalysis, and discloses a bio-enzyme catalysis process for reducing agent modification, which comprises the following steps: S1, enzyme solution preparation: mixing a bio-enzyme preparation with a buffer solution with the pH value of 5-8 to prepare an enzyme solution with the enzyme concentration of 0.5-5.0 g / L; the biological enzyme preparation is a compound enzyme composed of one or more of cellulase, hemicellulase and lignin degrading enzyme; s2, enzymolysis modification: mixing a carbonaceous reducing agent raw material with the enzyme solution prepared in the step S1 according to a mass ratio of 1: 5-1: 20, carrying out an enzymolysis reaction at 40-60 DEG C, and meanwhile, applying ultrasonic waves to carry out auxiliary treatment; the power of the ultrasonic waves is 100-500 W, and the frequency of the ultrasonic waves is 20-40 kHz And S3, solid-liquid separation: after the reaction is finished, separating the modified reducing agent from the enzymatic hydrolysate to obtain the solid modified reducing agent. Compared with the prior art, the method has the advantages that a biological enzyme catalysis process for modifying the reducing agent is provided, the energy consumption is low, the environment is protected, the cost is low, and the performance of the reducing agent can be remarkably improved.

One aspect of the invention relates to an oral health enzyme complex for the treatment of plaque and plaque bacterial biofilms and for the natural whitening of tooth enamel, comprising the combination of:-a sucrase enzyme, in particular a beta-2, 6 and / or beta-2, 1 bond capable of hydrolyzing sucrose; -a mixture of glucanohydrolases comprising a mutanase having excision activity and obtained from fungi, and a mutanase having excision activity and obtained from bacteria, in particular capable of hydrolyzing (1-3)-alpha-D-glucan and (1-6)-alpha-D-glucan; -a glycoside hydrolase enzyme, in particular a (1-6)-alpha-D glycosidic bond capable of hydrolyzing monosaccharide dimers and trimers; an oxide reductase, in particular capable of forming hydrogen peroxide from glucose and monosaccharide monomers; -a peroxidase, in particular capable of converting thiocyanates into hypothiocyanates in the presence of hydrogen peroxide.

The invention relates to the field of biological catalysis and enzyme engineering, and discloses a pH response type metal ligand-mutant metal dependent enzyme compound as well as a preparation method and application thereof. The compound is formed by compounding a mutant metal-dependent enzyme and a pH response type metal-ligand material, the mutant metal-dependent enzyme is obtained by directionally mutating amino acid residues in the metal-dependent enzyme, and the pH response type metal-ligand material is obtained by compounding the mutant metal-dependent enzyme and the pH response type metal-ligand material. The repulsive action of the amino acid residues on target metal ions in a metal ion transport channel of the metal-dependent enzyme is reduced or the amino acid residues and the target metal ions attract each other; the pH response metal-ligand material is formed by target metal ions and polydentate organic ligands through coordination, and the target metal ions can be complexed or released by adjusting pH. The efficiency that metal ions enter an enzyme activity center can be greatly improved, on-demand release and cyclic complexing of the metal ions are achieved, reutilization can be achieved through pH callback, the cost is effectively reduced, and excellent catalytic performance and environment friendliness are achieved.

The invention relates generally to the field of genome editing and provides DNA recombinases, which efficiently and specifically recombine genomic target sequences via the fusion of recombinase monomers. More specifically, the invention provides a method to generate a fusion protein for efficient and specific genome editing, comprising a complex of recombinases comprising at least a first recombinase enzyme, a second recombinase enzyme and at least one linker, wherein said first recombinase enzyme and said second recombinase enzyme specifically recognize a first half-site and a second half-site of an upstream target site and / or a downstream target site of a recombinase; wherein said first recombinase enzyme and said second recombinase enzyme are interconnected via a linker; and wherein said linker comprises or consists of an oligopeptide. The invention further relates to fusion proteins generated with this method. The invention also discloses designer-recombinases, which catalyze the inversion of a DNA sequence present in the int1h regions on the human X chromosome. The invention further relates to nucleic acid molecules encoding said DNA recombinases and fusion proteins, as well as to the use of said fusion proteins, DNA recombinases and nucleic acid molecules in a pharmaceutical composition.

The invention discloses a GSPT1 molecular glue degradation agent compound and application thereof, and particularly provides a compound shown as a formula I or pharmaceutically acceptable salt thereof. The invention aims to solve the technical problem that a high-selectivity and high-efficacy GSPT1 molecular glue degradation agent which can be used by GSPT1 dependent tumor patients is absent clinically at present, and provides a GSPT1 molecular glue degradation agent compound. The compound disclosed by the invention can be combined with an E3 ligase CRBN compound and change the specificity of the compound so as to induce ubiquitination and degradation of a compound related protein GSPT1, is novel in structure and has high selectivity and high degradation efficacy on the GSPT1 protein.

The invention relates to a pectin methylesterase dipeptide inhibitor design method, which comprises: carrying out virtual screening treatment on a candidate dipeptide compound through a molecular docking algorithm to obtain a candidate dipeptide inhibitor; analyzing a three-dimensional structure result of molecular docking of the candidate dipeptide inhibitor and the pectin methylesterase, and constructing an interaction model of the dipeptide inhibitor and the enzyme compound; performing stability analysis on the interaction model by using a molecular dynamics simulation algorithm to obtain a stability analysis result; and calculating the binding energy of the dipeptide inhibitor and the enzyme compound according to the stability analysis result, and screening the candidate dipeptide inhibitors according to the binding energy to obtain the pectin methylesterase dipeptide inhibitor. The screening efficiency can be improved, and the experiment cost can be reduced.

The invention discloses a self-assembly double-enzyme compound, a preparation method and application of the self-assembly double-enzyme compound in synthesis of a prednisone intermediate, and the self-assembly double-enzyme compound is obtained by fusing 11 alpha-hydroxylase and 1, 2-dehydrogenase on the basis of a SpyCatcher / SpyTag system. According to the method, a self-assembled double-enzyme compound of 11alpha-hydroxylase and 1, 2-dehydrogenase is constructed through a SpyCatcher / SpyTag system, 17alpha-hydroxyprogesterone is taken as a substrate, double-enzyme cascade one-step catalytic reaction is realized, the reaction process is simplified, and separation and purification operation of an intermediate product is omitted. The self-assembled double-enzyme compound constructed by the invention forms a high-efficiency substrate channel, accelerates the rapid transfer of an intermediate from a catalytic activity center of a first enzyme to a catalytic activity center of a second enzyme, improves the catalytic efficiency, is suitable for industrial application, and has a conversion rate of 99.6%.

The invention relates to a method and a process for synthesizing adenosine triphosphate through in-vitro multi-enzyme cascade catalysis and biological preparation of an enzyme compound, which are characterized in that the novel enzyme cascade catalysis for synthesizing ATP (adenosine triphosphate) adopts single enzyme protein for full-process catalysis, the related enzymes comprise adenosine kinase (EC 2.7. 1.20), adenylate kinase (EC 2.7. 4.3) and ADP (adenosine diphosphate) kinase (EC 2.7. 4.1), each enzyme is connected by using a connecting protein Linker, and the whole catalytic reaction process is as follows: (1) converting adenosine under the catalysis of adenosine kinase to generate AMP (adenosine monophosphate); (2) converting the AMP into ADP under the catalysis of adenylate kinase; and (3) converting the ADP into ATP under the catalysis of ADP kinase. And the contact path between the enzyme and the substrate is shortened, so that the product catalyzed in the previous step can quickly enter the enzyme catalytic reaction in the next step. The enzyme protein is immobilized, so that the application frequency of the enzyme protein is increased, and the production cost is greatly reduced.

The invention discloses an engineering bacterium and a method for synthesizing 3 '-sialic acid lactose based on an RIAD-RIDD self-assembled multienzyme compound. The present invention provides a recombinant microorganism for the production of 3 '-sialic acid lactose wherein the recombinant microorganism has (a) enhanced enzymatic activity of CMP-N-acetylneuraminic acid synthetase and / or expression level of coding gene thereof compared to a wild-type microorganism or an original strain; (b) enhancing the enzymatic activity of the alpha-1, 3-fucosyltransferase and / or the expression level of the encoding gene of the alpha-1, 3-fucosyltransferase; and (c) the CMP-N-acetylneuraminic acid synthetase and the [alpha]-1, 3-fucosyltransferase enzyme form a complex. The recombinant microorganism provided by the invention can improve the intermediate channelization efficiency and yield, has improved substrate utilization rate and is strong in universality.

The invention relates to the field of natural product chemistry, sugar chemistry and biological medicine, and particularly provides glycosaminoglycan and / or a series of derivatives thereof as well as a preparation method and application of the glycosaminoglycan and / or the series of derivatives thereof, the glycosaminoglycan is extracted from starfish glabrata, and the preparation method comprises the following steps: extracting by adopting a papain combined alkaline hydrolysis method; the invention discloses an ALHX-2M series derivative and application of the ALHX-2M series derivative in prevention or treatment of thrombotic diseases, and the ALHX-2M series derivative is obtained by performing separation and purification through ion exchange column chromatography and molecular exclusion column chromatography to obtain ALHX-2M, and further performing deacetylation, deamination and depolymerization treatment and gel column chromatography separation and purification on the ALHX-2M series derivative and the application of the ALHX-2M series derivative in prevention or treatment of thrombotic diseases. The glycosaminoglycan and the derivative thereof take an endogenous blood coagulation factor X enzyme complex (FXase) as an action target, the anticoagulation activity and the antithrombotic activity of the glycosaminoglycan and the derivative thereof are superior to those of low-molecular-weight heparin, the bleeding risk under the equivalent antithrombotic dosage is remarkably reduced, and the glycosaminoglycan and the derivative thereof can be used for medicines or functional foods for preventing and treating thrombotic cardiovascular and cerebrovascular diseases.

The invention discloses a saccharomyces cerevisiae engineering bacterium for efficiently synthesizing mogroside V based on peroxisome and lipid droplet double-cell area compartment and a construction method of the saccharomyces cerevisiae engineering bacterium. According to the construction method disclosed by the invention, squalene epoxidase ERG1 is anchored on the surface of a peroxisome membrane and in lipid droplets respectively; the ERG1 and downstream metabolic enzymes such as cucurbitadienol synthetase, cyclic epoxy hydrolase, cytochrome P450 enzyme, cytochrome P450 enzyme reductase, phosphoglucomutase, alpha-phosphoglucomutase, uridine diphosphate glucose pyrophosphorylase and glycosyltransferase are assembled into a multi-enzyme complex by adopting a multi-enzyme complex directional assembly strategy; the method not only enhances the directionality of metabolic flux, but also improves the catalytic efficiency of the enzyme and reduces the generation of byproducts, thereby realizing efficient synthesis of MG-V.

This disclosure relates to mRNA therapy for the treatment of maple syrup urine disease (MSUD). mRNAs for use in the invention, when administered in vivo, encode branched chain α-ketoacid dehydrogenase complex (BCKDC) E1α, E1β, or E2mRNA therapies of the disclosure increase and / or restore deficient levels of E1α, E1β, or E2 expression and / or BCKDC activity in subjects. mRNA therapies of the invention further decrease abnormal accumulation of branched chain amino acids associated with deficient BCKDC activity in subjects.

The invention discloses a kit for catalyzing Friedel-Crafts acylation reaction through an enzyme method, application and a preparation method. The kit for catalyzing the Friedel-Crafts acylation reaction by the enzyme method comprises 1) a Friedel-Crafts acylation reaction buffer solution and 2) a thiolase complex MucABC or a functional fragment and a variant thereof. The kit for catalyzing the Friedel-Crafts acylation reaction does not depend on coenzyme A compounds as acyl donors, and can realize regional specific C-acylation modification on various lactam compounds by taking acyl phenyl esters with different chain lengths as the acyl donors under mild conditions; a novel green and efficient biological catalysis tool is provided for Friedel-Crafts acylation reaction, and the method has important value and significance for drug synthesis and chemical production.

An enzyme has an activity that converts uridine in RNA to cytidine. A complex includes: the enzyme; and a sequence recognition module that allows the enzyme to act specifically on uridine that has occurred in mRNA due to mutation.