84 results about "Enzyme synthesis" patented technology

The invention discloses application of copper-amino acid nano enzyme in preparation of anti-inflammatory drugs, and relates to the technical field of biomedical new materials, amino acids comprise glycine, arginine, histidine, threonine, phenylalanine and cysteine; the ratio of the amino acid to the copper ions is 1: (0.5-5), and the synthesis temperature of the copper-cysteine nano enzyme is 25-125 DEG C. The copper-amino acid nano-enzyme library established by the invention has efficient hydroxyl free radical, superoxide free radical and hydrogen peroxide scavenging activity; wherein the copper-cysteine nano-enzyme shows the highest enzymatic activity, and shows low toxicity and good biocompatibility in both the cell level and the animal level; meanwhile, the copper-cysteine nano-enzyme also shows anti-inflammatory activity and anti-oxidative stress activity, can remarkably improve cell inflammation and body inflammation, relieves and treats dextran sodium sulfate induced mouse ulcerative colitis, and can be further applied to preparation of drugs for treating inflammatory bowel diseases.

The invention belongs to the technical field of biochemical engineering, and discloses a production method for improving the content of odd-chain fatty acid in grease by modifying schizochytrium limacinum and optimizing a substrate and application of the method. According to the method, schizochytrium limacinum HX-308 serves as a receptor strain, high-yield odd-chain fatty acid engineering bacteria are obtained through gene overexpression and inhibition, and the high-yield odd-chain fatty acid is obtained. Accumulation of a propionyl CoA precursor pool for synthesizing odd-chain fatty acid is improved, endogenous methylmalonyl-CoA mutase coding gene MUT is inhibited, so that a competitive pathway of the odd-chain fatty acid is inhibited, and then through substrate optimization, the content of fermented grease and the content of the odd-chain fatty acid are obviously improved. The expression of related enzymes in schizochytrium limacinum is adjusted through a genetic engineering strategy to improve the odd-chain fatty acid synthesis, the synthesis and utilization of the key intermediate propionyl coenzyme A in the odd-chain fatty acid synthesis process are increased, the metabolic pathway of the propionyl coenzyme A is inhibited, and substrate optimization is assisted to improve the odd-chain fatty acid yield.

The present disclosure relates to methods of using single-stranded RNA ligase for synthesis of oligonucleotides, including oligonucleotides containing a conjugate moiety. In some embodiments, the present disclosure further provides recombinant single-stranded RNA ligases, polynucleotides encoding the recombinant single-stranded RNA ligases, and compositions of the single-stranded RNA ligases.

The invention discloses a black fungus AMY protein and a coding gene and application thereof, and belongs to the technical field of protein engineering. The invention provides a black fungus AMY protein as well as an encoding gene and application thereof in order to excavate a key enzyme gene of a starch sucrose metabolic pathway in black fungus, and discovers and verifies that the black fungus AMY protein and the encoding gene thereof play a key regulation and control role in inducing synthesis of black fungus alpha-amylase through transcriptome sequencing. A solid foundation is laid for deeply exploring the function of a key gene of a starch sucrose metabolic pathway.

The application discloses a kind of high-activity polyphenol oxidase and preparation method thereof, belong to nano mimic enzyme synthesis technical field.The polyphenol oxidase of the application is with copper ion as active center, 2-methyl imidazole and guanylate disodium as organic ligand.Preparation method thereof includes the following steps: guanylate disodium solution is added to copper chloride solution under stirring condition, to obtain first mixed solution;First mixed solution is slowly added to 2-methyl imidazole solution under stirring condition, to obtain second mixed solution;Second mixed solution is centrifuged after ultrasonic at room temperature, to obtain brown precipitate, and the polyphenol oxidase is prepared by washing and drying brown precipitate.The mimic enzyme prepared by the application has higher catalytic performance, better stability and feasibility of large-scale preparation compared with natural enzyme, and will have broad application prospect in the field of biosensor.

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.

The invention provides non-naturally occurring microbial organisms containing enzymatic pathways and / or metabolic modifications for enhancing carbon flux through acetyl-CoA. In some embodiments, the microbial organisms of the invention having such pathways also include pathways for generating reducing equivalents, formaldehyde fixation and / or formate assimilation. The enhanced carbon flux through acetyl-CoA, in combination with pathways for generating reducing equivalents, formaldehyde fixation and / or formate assimilation can, in some embodiments, be used for production of a bioderived compound. Accordingly, in some embodiments, the microbial organisms of the invention can include a pathway capable of producing a bioderived compound of the invention. The invention still further provides a bioderived compound produced by a microbial organism of the invention, culture medium having the bioderived compound of the invention, compositions having the bioderived compound of the invention, a biobased product comprising the bioderived compound of the invention, and a process for producing a bioderived compound of the invention.

The invention provides a synergistic treatment pharmaceutical composition for preventing and treating recurrence after hepatocellular carcinoma thermal ablation and application of the synergistic treatment pharmaceutical composition. Aiming at malignant tumor progression driven by ACSS3 gene silencing after thermal ablation, an epigenetic editing technology is innovatively combined with metabolite supplementation, and a preparation for specifically activating or promoting ACSS3 gene expression and sodium propionate are cooperatively used, so that ACSS3 can be remarkably reactivated or expression can be promoted in a xenograft model derived from a cell line and a patient; the compound can be used for recovering propionyl-coenzyme A synthesis and normal propionic acid metabolism and inhibiting fatty acid beta-oxidation, so that the growth and metastasis of post-ablation hepatocellular carcinoma (HCC) are effectively blocked, and the limitation of a single therapy is overcome. A clearer mechanism target is provided for preventing recurrence after HCC thermal ablation, meanwhile, the prepared targeting nano-liposome preparation has a synergistic effect, and as a brand-new treatment strategy, the defect that intervention on recurrence roots is insufficient in the prior art is overcome.

The invention belongs to the technical field of biochemistry, and particularly relates to an efficient enzymatic synthesis method of holothurian glycosaminoglycan. According to the invention, a high-efficiency enzymatic synthesis approach of the sea cucumber glycosaminoglycan is established, sulfated fucose and chondroitin sulfate are used as raw materials, and the sea cucumber glycosaminoglycan with anticoagulant activity is efficiently prepared by using the alpha 1, 3-fucosyltransferase mutant and L-fucose kinase / GDP-fucose pyrophosphorylase and adopting a one-pot multi-enzyme synthesis strategy. And important technical support is provided for development of novel anticoagulant drugs.

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.

This invention discloses a method and system for controlling the preparation of low-temperature, high-efficiency straw-promoting microbial agents, relating to the field of microbial agent preparation process control technology. It utilizes multiple types of sensors to collect a microbial agent preparation dataset RAW(t) at time t, calculates and constructs a metabolic feature vector Vm(t) at time t, achieving real-time quantification of the macroscopic metabolic activity of microorganisms. Furthermore, it calculates the respiratory entropy Q at time t. R (t) and calculate the enzyme synthesis potential E at time t. S (t) and construct the diagnostic state vector Ds(t) at time t, thereby transforming external measurable parameters into a deep insight into the internal production potential of the microbial cells, objectively and dynamically diagnosing the fermentation process into different physiological stages, and generating a stage identifier ID(t) at time t, automatically executing the matching fine control strategy AIR, ensuring that optimal regulation is automatically adopted at each stage, thereby significantly improving the quality, production efficiency and batch stability of the microbial agent product.

The invention discloses a plant source leuco-anthocyanidin dioxygenase mutant and application thereof in synthesis of chiral drug intermediates, and belongs to the technical field of enzymology engineering. According to the invention, leuco-anthocyanidin dioxygenase (LDOX) derived from arabidopsis thaliana is modified through site-specific mutagenesis, an F304L / T239S double mutant (LDOXLS) is obtained, the (R)-phenylglycine analogue can be synthesized with the yield of 1.69 g / L, the yield of 81% and the enantiomeric excess ratio of 97: 3, and different substituent derivatives of the (R)-phenylglycine analogue can be generated. Compared with the traditional method, the method for synthesizing the (R)-phenylglycine analogue and the derivative thereof by using the enzyme has the advantages of wider substrate range, higher stereoselectivity, more environment-friendly requirement, simplicity and convenience in operation and the like.

The invention relates to the technical field of gene engineering, in particular to high-activity peptide ligase and application thereof in synthesis of tilpotide. The high-activity peptide ligase provided by the invention shows catalytic efficiency far higher than that of an existing enzyme in a specific ligation reaction aiming at an acyl donor and an acyl receptor of a precursor fragment of the tilpoitide; by systematically transforming a host strain, the high-activity peptide ligase which can be secreted and expressed at high level and high fidelity and has correct folding and complete activity is obtained, and meanwhile, the degradation of a product is reduced to the maximum extent; and an optimal expression element combination of the high-activity peptide ligase is obtained through high-throughput optimization of an expression system. The method is finally integrated into a complete new process for synthesizing the tilpoitide by the chemical enzyme method, which can realize kilogram-level production, and is suitable for popularization and application.

The invention discloses an enzymatic synthesis method of a melogabalin intermediate and related alcohol dehydrogenase, racemic 3-ethyl bicyclo [3.2. 0] hept-3-ene-6-ketone is used as a substrate, a genetically engineered bacterium for producing alcohol dehydrogenase is used as an enzyme catalyst, a genetically engineered bacterium for producing formate dehydrogenase, ammonium formate and coenzyme are matched, an enzymatic reaction is carried out under a stirring condition, and the melogabalin intermediate is obtained. And carrying out purification and separation on a reaction product to obtain the melogabalin intermediate (1R, 5S)-3-ethyl bicyclo [3.2. 0] hept-3-ene-6-ketone. The method has the advantages of simple reaction, few steps, mild reaction, less pollution and the like, and has great industrial application potential and commercial value.

This invention discloses a method for designing optimal strains for product synthesis using an enzyme-constrained model combined with an evolutionary algorithm, comprising the following steps: Step S1, solving the enzyme-constrained model with the objective function of maximizing the specific growth rate of the strain to obtain the simulated metabolic flux of the wild-type strain; Step S2, performing dimensionality reduction analysis and labeling on genes directly related to enzyme synthesis within the model by solving a series of flux balance analysis problems with fixed biomass synthesis rates; Step S3, predicting the yield of single-target editing; Step S4, using a genetic algorithm to search for combined targets after dimensionality reduction, adjusting relevant parameters to obtain the optimal gene editing combination strategy; Step S5, repeating the experiment to verify the stability of the algorithm. This invention utilizes genetic algorithms and enzyme-constrained models to discover non-intuitive gene editing combination strategies, providing a new method for constructing efficient microbial cell factories for synthetic chemicals, and demonstrating the potential of applying heuristic algorithms to strain optimization design.

The invention discloses streptomyces hygroscopicus ascomycetes subspecies and an obtaining method thereof. Regulating genes fkbN, fkbR1 and fkbR2 on an ascomycin biosynthesis gene cluster of the streptomyces hygroscopicus ascomycetes subspecies ATCC 14891 and a gene fkbS participating in synthesis of an ascomycin precursor ethyl malonyl coenzyme A are subjected to biosynthesis to obtain the streptomyces hygroscopicus ascomycetes subspecies. And carrying out overexpression on a regulatory gene SH03686 of a leucine response regulatory protein family (Lrp) outside the gene cluster, and screening an ascomycin high-yield strain. According to the invention, after combined overexpression of genes fkbS and fkbN and overexpression of a gene fkbR2 are carried out by a strain SN9 and a strain R2-17 in starting bacteria, the transcriptional level of regulatory genes (fkbN, fkbR2) on an ascomycin biosynthesis gene cluster and related genes participating in synthesis of precursors ethyl malonyl coenzyme A (fkbS, fkbE), piperidine acid (fkbL) and DHCHC (fkbO) are improved, and finally, the ascomycin yield is improved.

The invention discloses serratia marcescens for producing terpenoids and application of the serratia marcescens, and belongs to the technical field of bioengineering. According to the invention, part of genes, such as acetoacetyl-coenzyme A thiolase, 3-hydroxy-3-methylglutaryl-coenzyme A synthetase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase and phosphomevalonate decarboxylase, in a haloarchaea MVA pathway are integrated in chromosomes of a serratia marcescens host, and mevalonate kinase is expressed on plasmids, so that the recombinant serratia marcescens is obtained. Meanwhile, competitive genes are knocked out, and a universal terpenoid production strain is constructed. Synthetases of other terpenoids are further introduced into the universal terpenoid production strain and are used for synthesizing more terpenoids.

Disclosed herein are polynucleotides comprising sequences encoding coenzyme M synthase (ComF) linked to a heterologous regulatory element and methods of using the same. The polynucleotides may comprise synthetic operons comprising additional sequences encoding enzymes, e.g., a taurine-pyruvate aminotransferase, a sulfoacetaldehyde acetyl transferase, or a sulfopyruvate decarboxylase. Also disclosed herein are recombinant prokaryotic cells, e.g., recombinant bacterial, e.g., E. coli, or archaeal cells, e.g., Methanosarcina acetivorans with improved tolerance to oxidative stress.

The present application relates to the field of recombinant enzyme catalytic engineering technology, and provides a kind of intracellular expression lipase Ndbn and its related recombinant engineering bacteria.The lipase described in the present application has the characteristics of organic solvent resistance, can use aniline and ester as substrate in organic solvent, and enzyme synthesis amide compounds.Compared with existing lipases, lipase Ndbn shows high catalytic activity to weak nucleophile aniline, has a wide substrate spectrum, the aromatic amide catalyzed by it can be used as an intermediate of drug, avoids the shortcomings of high temperature and use of toxic reagent in the process of chemical synthesis of amide, has high atom economy, and meets the concept of green manufacturing.

The invention relates to the field of synthesis of natural dyes by genetic engineering and a biological method, in particular to a recombinant engineering bacterium for synthesizing N-acetyl blue as well as a construction method and application of the recombinant engineering bacterium. According to the invention, Escherichia coli BL21 (DE3) is used as an initial strain for modification; the method comprises the following steps: by using a genetic engineering bacterium as a template, expressing an ornamental blue synthetase encoding gene bpsA, a 4 '-phosphoric acid pantetheinyl transferase encoding gene entD, a glutamine synthetase encoding gene glnA, a glutamate dehydrogenase encoding gene gdhA, an acetyl-coenzyme A synthetase encoding gene acs and N-acetyltransferase encoding genes from different sources, and constructing to obtain a recombinant engineering bacterium. The recombinant engineering bacteria take sodium acetate as a substrate, and the sodium acetate not only meets the growth requirements of cells, but also can provide sufficient precursor substances. After the recombinant engineering bacterium constructed by the invention is subjected to shake flask fermentation culture for 72 hours, the purity of the N-acetyl blue can be up to 99% or above, and the recombinant engineering bacterium has the potential of industrial application.

The utility model belongs to capric acid glyceride production equipment technical field, concretely relates to a kind of enzyme synthesis capric acid glyceride reaction kettle, including reaction kettle, agitator, it is characterized by: reaction kettle top is equipped with the quick sampling mechanism installed by tank top flange one, tank top flange two, quick sampling mechanism includes the pipeline system of venturi, suction tube, connecting pipe, wherein the top end of suction tube is connected to venturi throat, the bottom end of suction tube is inserted into the inside of reaction kettle, venturi two ends are connected nitrogen valve and backwash valve respectively, backwash valve connects reaction kettle inside, connecting pipe connects sample release pipe, sample release valve is installed on sample release pipe, nitrogen valve is connected with nitrogen pipeline.The flow of nitrogen in venturi safely and conveniently takes out reaction liquid, effectively improves sampling convenience and safety;Reaction liquid can be circulated in quick sampling mechanism and reaction kettle to avoid sample cross-contamination.

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.

The invention belongs to the technical field of enzyme engineering and biology, and discloses a nitrilase mutant as well as a coding gene, a recombinant vector, a recombinant gene engineering bacterium, an enzyme preparation and application thereof. The nitrilase mutant is obtained by carrying out single-point mutation or multi-point mutation on 8th, 17th, 35th or 150th amino acids of an amino acid sequence of the nitrilase as shown in SEQ ID NO.2. The invention further discloses a preparation method of the nitrilase mutant. Compared with wild type Gi-Nit, the nitrilase mutant disclosed by the invention has the advantages that the activity is improved by 32 times, the yield is improved by 15.8 times, and a foundation is laid for synthesizing 2-(4-nitrophenyl) butyric acid by an industrial enzyme method.

The present invention relates to a recombinant microorganism for producing 1,3-propanediol and a method for producing 1,3-propanediol using same. More specifically, the present invention provides a recombinant microorganism having an improved ability to produce 1,3-propanediol through overexpression of a transaldolase-encoding gene (tal), a succinyl-CoA synthetase-encoding gene (sucCD), a cystathionine γ-synthase-encoding gene (metB), an aspartate ammonia lyase-encoding gene(aspA), an aspartate aminotransferase-encoding gene (aspB), or a functional fragment thereof, and a method for producing 1,3-propanediol using same.

The present application relates to the technical field of enzyme synthesis of target compounds, and particularly relates to a method for preparing diglyceride vegetable oil by enzyme method, which comprises the following steps: mixing vegetable oil and glycerol according to a mass ratio of 3:1, then adding 5%-10% of a composite enzyme resin to the mixed liquid in terms of the mass of the vegetable oil, oscillating and treating at 40-60 DEG C for 8-16h, and then performing solid-liquid separation to obtain the diglyceride vegetable oil; wherein the composite enzyme resin takes macroporous resin AB-8 as a carrier, the carrier pores are fixed with partial glyceride lipase, the carrier exterior is coated with a gel layer, and the gel layer is dispersed with Candida antarctica lipase A. The method for preparing the diglyceride vegetable oil by enzyme method provided by the present application shortens the original method for obtaining the diglyceride vegetable oil crude product in steps to one step, belongs to a one-pot operation, and the diglyceride content of the diglyceride vegetable oil produced finally is high.

The invention relates to the technical field of cyclopeptide drug synthesis, in particular to a method for synthesizing mirror cyclopeptide by using OaAEP1 enzyme under the assistance of thioester, which comprises the following steps: firstly, synthesizing a linear mirror cyclopeptide thioester precursor by using an N-fluorenylmethoxycarbonyl (Fmoc) solid-phase polypeptide synthesis method, and then generating the mirror cyclopeptide from the thioester precursor under the mediation of the OaAEP1 enzyme. According to the invention, the mirror image ring melittin (D-cMelittin) is synthesized by using OaAEP1 enzyme under the assistance of thioester for the first time, and the mirror image ring melittin (D-cMelittin) has high protease stability and good antibacterial activity.

The invention relates to the technical field of enzyme catalysis, and particularly discloses a method for biologically synthesizing succinic acid in vitro by taking L-glutamic acid as a raw material, which constructs a new path for synthesizing L-glutamic acid-3-cyanopropionic acid-succinic acid by a full-biological enzyme method for the first time. A green conversion chain from cheap and bulk amino acids to a high value-added biodegradable plastic precursor is broken through, an L-glutamic acid resource with leading productivity is in butt joint with a poly (butylene succinate) (PBS) industry chain in shortage in the market, and the whole process of finally synthesizing succinic acid from L-glutamic acid through a 3-cyanopropionic acid intermediate is realized. According to the process, full-water-phase and extremely-mild one-pot method or multi-enzyme coupling conversion is realized. The two-step reaction is carried out in a nearly neutral water phase system, and the compatibility of enzyme is excellent, so that the process flow is greatly simplified, the equipment investment and energy consumption are reduced, common harmful byproducts in chemical synthesis are reduced from the source, and the biological safety and extremely high purity of the product succinic acid are ensured.

The application provides a genetically engineered bacterium with high yield of dihydroquercetin, a preparation method and application thereof, relates to the technical field of genetic engineering, and comprises a malonyl coenzyme A synthesis module, a tyrosine deaminase expression module, a naringenin synthesis module and a dihydroquercetin post-modification module. The enzymatic property of the fusion protein is increased by about 50% of the enzyme activity before fusion, the activity of converting naringenin into dihydroquercetin is greatly improved, and finally the yield of dihydroquercetin expressed in cyanobacteria is greatly improved. Meanwhile, BauA, MCR-C, Sam8, 4CL, CHS, CHI and expression fusion protein N33'H are expressed in cyanobacteria. The yield of dihydroquercetin is greatly increased under seawater culture conditions. The technical problem that the yield of biosynthetic dihydroquercetin is low and cannot meet the requirements of industrial application in the prior art is solved.

The invention discloses a pesticide synergist containing platinum nano enzyme and a preparation method of the pesticide synergist. The pesticide synergist comprises nano silicon dioxide, the platinum nano enzyme, chitosan and a surfactant. The preparation method comprises the following steps: S1, synthesizing hollow dendritic nano silicon dioxide; s2, synthesizing platinum nano enzyme; and S3, synthesizing the pesticide synergist containing the platinum nano enzyme. According to the invention, the preparation conditions of the platinum nano enzyme are optimized, and the production requirements are reduced. The nano-enzyme agricultural synergist prepared by the method disclosed by the invention can be used for flight control, and the labor cost is reduced.