19 results about "Enzymic hydrolysis" patented technology

The invention discloses a stipa purpurea tissue culture and rapid propagation method and a special culture medium. The method comprises the following steps: (1) selecting and pretreating an explant; (2) primary culture; (3) subculture; (4) rooting culture; and (5) seedling hardening and transplanting. Through an optimized explant disinfection process, a culture medium system based on N6, a specific plant growth regulator combination and addition of enzyme hydrolysis casein in a rooting stage, a breakthrough of a stipa purpurea tissue culture and rapid propagation technology is achieved, the pollution rate is lower than 5%, the multiplication coefficient reaches 4.2 or above, the rooting rate exceeds 93%, the transplanting survival rate reaches up to 89%, and the tissue culture and rapid propagation technology has the advantages that the tissue culture and rapid propagation efficiency is high. The whole rapid propagation period only needs 60-70 days, and efficient and reliable technical support is provided for large-scale and standardized production and ecological restoration application of stipa purpurea.

The invention provides a method for treating PET (Polyethylene Terephthalate) and cotton blended fabric waste. The method comprises an ultrasonic pretreatment step and an enzyme hydrolysis step, according to the method, PET and cotton in the fabric waste are separated from each other. The invention further provides a system for implementing the method.

The invention discloses a polypeptide compound and application thereof in preparation of a product for inhibiting ATP enzyme hydrolysis activity. The invention provides a polypeptide which is any one of the following: A1, the amino acid sequence of the polypeptide is sequence 1; a2, tag protein is added to the tail end of the polypeptide shown in A1, and the fused polypeptide is obtained. According to the method, multi-round progressive sequence optimization is performed by focusing a prototype peptide targeting 2C, simulating comprehensive utilization of a virtual neural network training model and actually-measured biochemical function evaluation in multiple aspects such as enzymatic inhibition and binding force evaluation, and a series of candidate peptides with relatively high comprehensive evaluation and shortened sequences are obtained; and a structural basis is provided for the design of a new round of antiviral peptidomimetic small molecules. Finally, candidate peptides are obtained through the research, and theoretical basis and technical support are provided for development of ATP enzyme inhibitors.

The invention discloses a beta-N-acetylglucosaminidase mutant obtained based on random mutation and a preparation method of the beta-N-acetylglucosaminidase mutant. On the basis of an error-prone PCR (Polymerase Chain Reaction) technology, a mutant BsNagZ-14 (C17R / K209T / T436S) and a mutant BsNagZ-17 (E358G) of the beta-N-acetylglucosaminidase of the bacillus subtilis are obtained in the invention. The two mutants keep high enzyme activity within the pH range of 3-10, the thermal stability of BsNagZ-14 is better, 98% of enzyme activity (90% of original enzyme) is kept after the temperature is kept at 40 DEG C for 3 hours, 83% of enzyme activity is kept after the temperature is kept at 60 DEG C for 3 hours, and the parent is attenuated to 23% of the initial enzyme activity. The mutants BsNagZ-14 and BsNagZ-17 are connected with a carrier pET20b (+) and are transferred into escherichia coli BL21 (DE3), the extracellular highest enzyme activities of the two recombinant bacteria are 1.140 U / mL and 1.136 U / mL respectively, and the extracellular highest enzyme activities of the two recombinant bacteria are improved by 16.32% and 15.91% respectively compared with those of control group bacteria expressing NagZ. The beta-N-acetylglucosaminidase can be externally tangent to the chitosan oligosaccharide, so that the hydrolysis effect of the chitinase on the chitin is favorably improved. The beta-N-acetylglucosaminidase mutant has wide pH and temperature adaptability, can be adapted to chitinase with different enzymatic characteristics, and provides convenience for carrying out double-enzyme hydrolysis on chitin.

This invention discloses a protease-responsive GPCR receptor and its applications. The protease-responsive GPCR receptor of this invention comprises an extracellular protease recognition domain, a transmembrane domain, and an intracellular signaling domain; the extracellular protease recognition domain contains a peptide hydrolyzed by the protease; the intracellular signaling domain contains an HCV enzyme hydrolyzed peptide, a nuclear localization sequence (NLS), a Cas protease, and a coupled nuclear transcription factor. This protease-responsive GPCR receptor can induce endogenous dCas9 assembly into the nucleus and activate the chimeric antigen receptor signaling pathway in natural killer cells in response to extracellular proteases, providing a new solution for developing novel precision immunotherapy.

The invention belongs to the field of bioengineering and solid waste resource utilization, and relates to a kitchen waste pretreatment method for propionic acid fermentation, a special microbial inoculum and a fermentation assistant.The kitchen waste pretreatment method for propionic acid fermentation comprises the steps that kitchen waste is crushed into particles of 2-5 mm, the particles are mixed with water according to the solid content of 40%-80%, the mixture is heated at the temperature of 50-80 DEG C and then filtered, and a mixture is obtained; filtering to obtain filtrate; performing oil-water separation on the filtrate to obtain a water phase and an oil phase; adding 1%-10% (v / v) of the obtained oil phase back into the water phase, and emulsifying at the rotating speed of 800-1000rpm; sequentially carrying out enzymolysis treatment of lipase hydrolysis, alpha-high-temperature amylase hydrolysis and glucoamylase hydrolysis on the emulsified treatment liquid; after impurity adsorption, enzyme deactivation is performed to obtain the propionibacterium fermentation substrate. The strain can be used for efficient substrate conversion, has convenient microbial inoculum application, optimizes the fermentation process, improves economic and environmental benefits, and has industrial application potential.

To provide a separation and discrimination method of a nucleus pulposus precursor cell, and application thereof.SOLUTION: Provided is a separation and discrimination method of a nucleus pulposus precursor cell, where a large number of tissue blocks are divided from a nucleus pulposus tissue, and the tissue blocks are left to stand and cultured in a culture dish after performing hydrolysis by enzyme. When cells are formed into the partition blocks on a bottom of the culture dish, a nucleus pulposus precursor cell is screened from the partition blocks by using a specific embryonic stem cell gene (e.g., Nanog, Oct-4, SOX2). Further, the method can screen a nucleus pulposus precursor cell having moving performance and / or mesenchymal stem cell properties by using a specific protein and a gene marker, and further can screen a nucleus pulposus precursor cell having anti-inflammatory performance. Finally, disclosed is application used for producing a pharmaceutical composition treating lower back pain by using the nucleus pulposus precursor cell obtained by the method, and a new therapy strategy to lower back pain is provided.SELECTED DRAWING: Figure 6

The invention provides a bio-based fabric dyeing process, which comprises: S1, enzyme hydrolysis treatment: placing a bio-based fabric in a compound enzyme solution, and carrying out heating treatment to obtain a carboxyl-containing bio-based fabric; s2, surface grafting: dipping the carboxyl-containing bio-based fabric obtained in S1 in a solution containing ethyl phosphate methacrylate and potassium persulfate, and carrying out a heating reaction to obtain an ethyl phosphate methacrylate grafted bio-based fabric; s3, cationic dye dyeing: dipping the ethyl phosphate methacrylate grafted bio-based fabric obtained in S2 into a cationic dye, and carrying out a heating reaction to obtain a dyed bio-based fabric; s4, post-treatment: performing water washing, soaping and drying on the bio-based fabric obtained in S3 to obtain the dyed bio-based fabric.

The present invention provides an enzymatic synthesis method for the immunosuppressant mycophenolic acid and its derivatives. This method utilizes a multi-enzyme, multi-cycle cascade reaction strategy to catalyze the conversion of the substrates 4-dienodecanoic acid-3-hydroxy-5-hydroxymethyl-6-methylphthalide or 4-dienodecanoic acid-3,5-dihydroxy-6-methylphthalide into mycophenolic acid and its derivatives. The multi-enzyme combination comprises an acyl-CoA ligase, an FAD-dependent oxidase, a 2,4-dienoyl-CoA reductase, an enoyl-CoA isomerase, a bifunctional enzyme with hydratase and dehydrogenase functions, and a thiolase; in addition, a CoA hydrolase and an oxygen-methyltransferase. This method, for the first time, achieves the in vitro enzymatic synthesis of (M)FDHMP-3C to MPA, fully elucidating the mycophenolic acid oxidative degradation synthesis pathway and the catalytic sequence of the catalytic enzymes.

This invention discloses a mutant S-transaminase and an immobilized enzyme-linked method for preparing (S)-3-aminobutyric acid (aminobutyric acid). The method involves site-directed mutagenesis of a wild-type thermostable Bacillus transaminase to obtain a mutant S-transaminase capable of efficiently converting ethyl acetoacetate to ethyl (S)-3-aminobutyric acid (aminobutyric acid), followed by esterase hydrolysis to obtain (S)-3-aminobutyric acid. This invention utilizes high-concentration substrates and isopropylamine and provides a solution suitable for continuous industrial-scale production to address the high concentration of byproducts.

The invention discloses soluble high-fiber sorghum flour and a preparation method thereof. The method comprises the following steps: soaking unhulled sorghum serving as a raw material in salt water to soften fibers; grinding, mixing with defatted soybean powder, and carrying out directional enzymolysis by using a compound enzyme, so that the content of starch is reduced, a fermentation carbon source is provided, and meanwhile, a lignin network is cracked to improve the dispersity and flavor; then bacillus licheniformis is inoculated for fermentation, fibers are softened by utilizing acid produced by metabolism of the bacillus licheniformis, protein and phytic acid are hydrolyzed by producing enzyme, and the solubility and flavor are remarkably improved; then fruit fibers are added, a fiber crystal structure is destroyed through homogenization, an emulsified network is formed, and the taste and the fineness are optimized; and finally, carrying out spray drying and fluidized bed granulation processes to obtain a finished product with excellent flowability, instant solubility and high fiber retention rate. While the dietary fiber nutrition of the whole sorghum grains is reserved to the maximum extent, the brewing property and sensory quality of the whole sorghum grains are effectively improved, and the market acceptability of the whole sorghum grains is improved.

The application relates to the technical field of organic water-soluble fertilizers, and discloses an amino acid organic water-soluble fertilizer and a preparation method thereof. The amino acid organic water-soluble fertilizer comprises the following components in weight: 40-50 parts of amino acid concentrate, 10-15 parts of organic matter, 20-25 parts of macroelement, 5-8 parts of medium element, 3-5 parts of polyaspartic acid, 4-6 parts of trace element, 0.4-0.6 parts of beneficial microbial agent, 2-4 parts of quaternary ammonium salt-based silicon dioxide, 0.2-0.3 parts of indole acetic acid and 0.5-1 part of synergist. The amino acid organic water-soluble fertilizer of the application uses waste animal and plant protein as raw material, has the advantages of environmental protection and low cost, and is superior to the commercially available product which depends on chemical synthesis of amino acid; through a multi-step enzymolysis (alkaline protease + acidolysis + papain / cellulase) process, the waste protein is efficiently converted into small molecule amino acid, the limitation of traditional single-enzyme hydrolysis is broken, more comprehensive free amino acid is released, and the plant absorption rate is significantly improved.

The invention discloses an active peptide for promoting bone growth, a composition as well as a preparation method and application of the active peptide. The active peptide for promoting bone growth comprises at least one of Leu-Arg, Leu-Leu, Phe-Leu and Ala-Phe. In order to overcome the defects of lack of traditional application and traditional Chinese medicine theory support, insufficient activity, large peptide fragment molecular weight and the like of existing food-borne polypeptides for promoting bone growth, the invention prepares polypeptides for promoting bone growth by utilizing an enzyme hydrolysis technology based on the traditional application of embryonated eggs, clarifies the structure of active polypeptides, performs quantitative analysis, and provides a basis for the food-borne polypeptides for promoting bone growth. Finally, the active peptide capable of promoting bone cell proliferation is provided, and the active peptide is natural, safe, free of toxic and side effects and capable of being taken for a long time.

The invention discloses a preparation method and application of a sturgeon caviar active extract, and provides the sturgeon caviar active extract, a cosmetic raw material, a composition of the cosmetic raw material and application of the sturgeon caviar active extract in preparation of cosmetics. Through a unique'three-removal 'pretreatment process, impurities, peculiar smell and part of grease are effectively removed, and optimal conditions are created for subsequent enzymolysis; the extract is good in stability, and the technical problem that the extract is easy to separate out in a cosmetic formula is solved; the effect is clear and obvious, and data support is sufficient; the final extract is subjected to comprehensive risk substance evaluation, and the safety is high.

The application belongs to the technical field of chitosan oligosaccharide preparation, and specifically discloses a low-polymerization-degree chitosan oligosaccharide as well as a preparation method and application thereof. First, chitosan and an organic acid solution are prepared into a chitosan colloidal solution; then, pectinase is used for pretreatment, and chitosanase is added for enzymolysis treatment to obtain an enzymolysis solution; after the enzymolysis solution is inactivated, centrifugal treatment and fractional alcohol precipitation are performed to obtain the low-polymerization-degree chitosan oligosaccharide with a polymerization degree of 2-4. The application adopts a double-enzyme hydrolysis strategy of pectinase and chitosanase, the pretreatment of pectinase can effectively improve the accessibility of the substrate, and a synergistic effect is generated with chitosanase, so that the degradation efficiency is significantly improved; the method has the advantages of mild process conditions and green environmental protection, and the prepared low-polymerization-degree chitosan oligosaccharide has significant anti-inflammatory activity.

The invention discloses low-purine and high-flavor bone soup and a preparation method thereof. According to the method, part of purine is removed through deep blanching, a core technology of performing enzymolysis by adopting a compound enzyme preparation of acid protease, nuclease and aminopeptidase is adopted, and the acid protease preferentially hydrolyzes collagen and muscle protein networks wrapping nucleic acid under a set pH value, so that the accessibility of a nucleic acid substrate is remarkably improved; then, nuclease specifically hydrolyzes the exposed RNA and DNA phosphodiester bond to degrade the RNA and DNA phosphodiester bond into nucleoside and purine / pyrimidine bases, and deep and efficient removal of purine is achieved from the source. Meanwhile, exonuclease can immediately trim the tail end of a peptide chain generated by protease hydrolysis, and accumulation of hydrophobic short peptides with bitter taste is effectively prevented. The final product not only retains the inherent mellow bone fragrance of the bone soup, but also has the excellent sensory quality of prominent delicate flavor, harmonious taste and long aftertaste.

The utility model provides a system for separating blended fabric through a biological method, the blended fabric is PET and cotton blended fabric, and the system comprises: (1) an ultrasonic generator for performing ultrasonic treatment on PET and cotton blended fabric waste; the enzyme hydrolysis reactor (2) is used for enabling the waste subjected to ultrasonic treatment to be subjected to an enzymolysis reaction; the system enables PET and cotton in the blended fabric waste to be separated from each other.

The present application aims to provide a new type of 5'-modified nucleoside based on selenophosphonate chemistry. Specifically, the present application designs a modified nucleoside compound, which can be efficiently introduced into the 5' end of an oligonucleotide chain by a solid-phase synthesis method. Compared with the prior art, the oligonucleotide synthesis step containing the modified nucleoside compound of the present application is more concise, can resist phosphatase hydrolysis, maintain the binding with Ago2 protein, and produce good gene silencing activity verified by in vitro activity. By utilizing the unique chemical properties of selenium atoms, the internalization behavior and cytoplasmic bioavailability of the modified oligonucleotide in extrahepatic tissue cells are potentially enhanced.

The invention discloses a method for synthesizing ppGpp by an enzymic method. The method comprises the following steps: 1) cloning from a genome of E.coli BL 21 (DE3), and expressing ppGpp synthetase RelA and pppGpp hydrolase GppA in vitro; and 2) by taking ATP and GTP as substrates, catalyzing and synthesizing pppGpp by utilizing ppGpp synthetase RelA, hydrolyzing the pppGpp by utilizing pppGpp hydrolase GppA to generate ppGpp, and preparing HPLC (High Performance Liquid Chromatography) to obtain the product ppGpp. According to the method, common compounds ATP and GTP are used as raw materials, ppGpp is synthesized through an enzymatic method, and compared with a chemical synthesis method or other enzymatic methods, the separation process is more economical, simpler and more efficient.