29 results about "Copper oxidase" patented technology

Methods and compositions for treating fibrosis (e.g., cardiac fibrosis), or other conditions associated with inflammation and / or fibrosis are provided. Methods can include administering a nucleic acid encoding a sialic acid-binding immunoglobulin-type lectin 9 (Siglec-9) protein to a subject in need of fibrosis treatment, where the Siglec-9 protein is expressed by the nucleic acid in regulatory T (Treg) cells in the subject. Methods and compositions can include Treg cells that overexpress Siglec-9 to enhance the ability of Treg cells to target cells expressing amine oxidase, copper containing 3 (AOC3), including fibroblasts or myofibroblasts. In some embodiments, the enhanced Treg cells are targeted to myofibroblasts (e.g., cardiac myofibroblasts) with elevated expression of fibrotic genes.

The invention discloses a Vinck Cellulomicrobe and application thereof in degradation of corn straw and improvement of soil fertility, and belongs to the field of agricultural microbial technology and solid waste resource utilization. The bacterial strain can secrete a plurality of hydrolase and oxidase including AA10 family lytic polysaccharide monooxygenase, AA2 peroxidase, GH3 and GH1 beta-glucosidase, GH16 / GH17 beta-glucosan endonuclease and AA1 multi-copper oxidase. The bacterial strain has the advantages that the bacterial strain can secrete a plurality of hydrolase and oxidase; fermentation liquor is applied to soil according to the straw mass ratio of 1: 5, and the corn straw degradation rate can be increased under the condition that the mass ratio of soil to straw is 100: 1. Meanwhile, the microbial agent can synergistically improve soil nutrients and increase the contents of rapidly available phosphorus, rapidly available potassium, alkali-hydrolyzable nitrogen, soluble organic carbon and total phosphorus in soil. According to the microbial agent, a straw lignocellulose structure is effectively cracked through a multi-enzyme system synergistic effect, and the dual effects of accelerating straw degradation and improving soil fertility are achieved.

The invention relates to the technical field of agricultural biology, in particular to a high-activity multi-copper oxidase mutant V286N and application thereof in mycotoxin degradation. The amino acid sequence of the multi-copper oxidase mutant is as shown in SEQ ID NO: 2. The multi-copper oxidase mutant provided by the invention has higher specific activity. Compared with wild type multi-copper oxidase, the multi-copper oxidase mutant MCO-V286N provided by the invention has the advantages that the specific activity is improved by 0.8 time, the degradation efficiency on mycotoxin zearalenone is improved by 0.4 time, the production and application cost is effectively reduced, and the multi-copper oxidase mutant MCO-V286N has important application value and technical significance in wide application of the multi-copper oxidase in the field of mycotoxin detoxification of food and feed.

The invention relates to the technical field of bioengineering, in particular to a process for continuously producing nicotine based on immobilized enzyme. The immobilized microbial agent comprises a composite carrier, an engineering strain and an immobilized auxiliary agent, in the process for continuously producing nicotine based on immobilized enzyme, amine oxidase of an engineering strain is subjected to site-specific modification, so that spatial structure deformation when the amine oxidase is combined with a carrier is reduced, and a three-dimensional network structure of hydroxyethyl chitosan-guar gum composite microspheres in the composite carrier is matched to provide an adaptive microenvironment; the activity loss of enzyme molecules caused by steric hindrance or structural change is reduced; in the immobilization auxiliary agent, gallic acid and an enzyme molecule sulfydryl form a coordinate bond to maintain an active center conformation, glutaraldehyde and polyethyleneimine enhance the binding stability through a crosslinking effect and charge regulation, and the maintaining effect of enzyme activity during continuous production is improved, so that the enzyme activity loss in the immobilization process is reduced, and the immobilization efficiency is improved. The stability of the enzyme in long-term continuous reaction is enhanced.

The invention discloses an application of a CuN3 / Cu3 artificial metalloenzyme in preparation of a zinc air battery and a self-powered sensing system. The enzyme simulates the active center of multi-copper oxidase, and shows high-performance catalytic activity on oxygen reduction reaction in a chemically favorable environment (half-wave potential E1 / 2 is equal to 0.86 V). The artificial metalloenzyme provided by the invention is verified in zinc air batteries (ZABs) and self-powered sensing systems (SPSSs) as an efficient ORR catalyst, and shows high power density, long-term stability and high sensitivity to a target substance glucose in a complex environment.

The invention discloses a multi-copper oxidase mutant sourced from Psychrobacter sp. And capable of efficiently degrading biogenic amine and application of the multi-copper oxidase mutant. According to the multi-copper oxidase mutant, the 183rd site of an amino acid sequence shown in SEQ ID NO.1 is mutated into histidine from phenylalanine, the 221st site of the amino acid sequence shown in SEQ ID NO.1 is mutated into aspartic acid from asparagine, the 261st site of the amino acid sequence shown in SEQ ID NO.1 is mutated into glutamic acid from glutamine, and the 292nd site of the amino acid sequence shown in SEQ ID NO.2 is mutated into arginine from lysine. Compared with wild type multi-copper oxidase, the multi-copper oxidase mutant disclosed by the invention has a good degradation effect on tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine, and the degradation rate on the spermidine and the spermine is increased to 100%, so that the multi-copper oxidase mutant has a good application prospect in food fermentation safety production.

The invention relates to a method for preparing a cycloheptatrienone derivative, and belongs to the technical field of bioengineering. The invention provides a method for preparing a cycloheptatrienone derivative, which is based on a framework editing strategy of biological catalysis and comprises the following steps of: performing biological catalysis on a substrate and an auxiliary substrate by using a phenol derivative as a substrate and nitroalkane as an auxiliary substrate and using multi-copper oxidase and / or a microorganism for producing the multi-copper oxidase as a catalyst to obtain the cycloheptatrienone derivative. The auxiliary substrate is used as a C1 unit to be inserted into the skeleton of the aromatic ring substrate for editing, so that the cycloheptatrienone derivative is generated. According to the method, multi-copper oxidase is used as a catalyst, nitroalkane which is simple, easy to obtain, safe and highly stable is used as a C1 unit, and a phenol derivative is successfully converted into the cycloheptatrienone derivative. According to the method, a single carbon insertion mediated skeleton editing biological catalysis strategy is constructed, the strategy breaks through the limitation of a traditional carbene insertion reaction, and the method has a great application prospect in skeleton modification in the field of chemical and biological catalysis.

The present invention provides recombinant Escherichia coli for producing glutarate, a construction method and use thereof. A double-plasmid recombinant bacterium is constructed through molecular biological means for co-expressing an aldehyde synthase (AAS) gene, an amine oxidase Mao (gene) and an aldehyde dehydrogenase (Glox) gene. The constructed expression plasmids are introduced into the Escherichia coli to reconstruct to obtain recombinant cells. A recombination strain for efficiently producing glutarate is obtained through amicillin resistance and kanamycin resistance combined plate screening. Efficient production of the glutarate is achieved by optimizing concentration of a substrate, cell concentration and a transformation temperature. L-lysine with a concentration of 30 g / L may be transformed into 19.65 g of glutarate through reactions for 30 h under transformation conditions that the cell concentration is 30 g / L, the pH value is 8 and 6 mM of NAD+ is additionally added, wherein a transformation rate may be 65.3%.

The invention discloses application of a multi-copper oxidase mutant in biogenic amine degradation, and belongs to the technical field of food biology. The amino acid sequence of the multi-copper oxidase mutant is shown as SEQ ID NO.1. The application is to degrade biogenic amines in aquatic products, and the biogenic amines comprise tryptamine, phenylethylamine, putrescine, cadaverine, histamine, octopamine, tyramine, spermidine and spermine. The multi-copper oxidase mutant disclosed by the invention not only can effectively reduce the content of biogenic amines in aquatic products and reduce the generation of reaction byproducts, but also can maintain the freshness and nutritional value of the aquatic products, and has remarkable application value in the fields of aquatic product processing and food safety.

This invention relates to the field of food labeling technology, and more particularly to a smart label assembly and its preparation method. A smart label assembly includes: a substrate layer, a color-developing layer, and a protective layer; the color-developing layer is fixed to the surface of the substrate layer, and the protective layer covers the surface of the color-developing layer; the color-developing layer includes: an amine oxidase-type color-developing area and a glucose oxidase-type color-developing area. When livestock and poultry meat spoils and produces volatile amines and glucose degradation products, these two types of substances are catalyzed by amine oxidase and glucose oxidase, respectively, to produce ammonia and hydrogen peroxide, which then react with indicators on the label to produce color changes. The two types of labels are used together for mutual verification, forming a rapid detection method for livestock and poultry meat spoilage using smart labels, which can directly determine the quality of livestock and poultry meat. This label is simple to prepare, low in cost, and suitable for fresh livestock and poultry meat packaging, allowing monitoring from the freshness stage.

This application relates to the field of air purification technology, specifically to a biomedical-based odor-removing and antibacterial air freshener, mainly composed of the following raw materials in parts by weight: 2.5-5.5 parts modified lysozyme microcapsules, 2.5-4.5 parts modified quaternary ammonium salt-β-cyclodextrin inclusion complex, 1.5-2.5 parts plasma amine oxidase, 2.0-3.5 parts compound plant extract, 0.2-0.3 parts citric acid, 0.5-0.6 parts lactic acid, and the balance being food-grade deionized water; the air freshener has a pH value of 6.0-8.5. The mist particle size is 10-50μm; the air freshener does not contain fragrances, alcohol, pigments, preservatives, chlorine disinfectants, hypochlorous acid, drugs, hormones and other irritating ingredients. All raw materials and modified carriers are food-grade and edible. Through a triple deodorization system of plasma amine oxidase, β-cyclodextrin and compound plant extracts, the ammonia removal rate is ≥95%, the odor suppression time is ≥100h, the product has a skin irritation level of 0, and the acute oral toxicity LD50 is >5000mg / kg, achieving a balance between mild and non-irritating properties and highly effective sterilization.

The present invention relates to novel bioprobes which are capable of binding to certain amine oxidase enzymes. These bioprobes are useful in methods of detecting and determining the concentration of certain amine oxidase enzymes in a sample as well as in methods for the quantitative assessment of inhibition of certain amine oxidases.

The invention discloses a CuN3 / Cu3 artificial metalloenzyme with precise coordination control and a preparation method of the CuN3 / Cu3 artificial metalloenzyme. The CuN3 / Cu3 artificial metalloenzyme with precise coordination control is reasonably designed by combining an excellent atomic structure and an electron coordination environment of a monatomic catalyst. The enzyme simulates the active center of multi-copper oxidase, and shows high-performance catalytic activity on oxygen reduction reaction in a chemically favorable environment (half-wave potential E1 / 2 is equal to 0.86 V). Advanced characterization and density functional theory calculation are utilized, the synergistic effect of CuN3-Cu3 sites in an enzyme sample mode is systematically studied, it is revealed that the change of the energy level of a d orbit reduces the potential barrier of the reaction, and the coupling interaction between CuN3 monatomic and a Cu3 cluster enhances adsorption and desorption of an oxygen intermediate, so that the reaction kinetics is improved.

The invention discloses preparation and application of alcohol-resistant amine oxidase LYYOBN2 capable of degrading biogenic amine, and belongs to the technical field of molecular biology. The invention provides the amine oxidase from bacillus, and the expression of the amine oxidase in escherichia coli is realized. According to the application of the prepared amine oxidase to degradation of the biogenic amine, the enzyme is added into fermented food, the biogenic amine can be effectively degraded in an environment containing ethanol or NaCl, an enzyme library for degrading the biogenic amine can be expanded, and the safety of the fermented food is further improved.

Method for degrading a vinyl polymer A method for degrading a vinyl polymer having an initial molar mass in order to form at least one degradation product having a final molar mass lower than the initial molar mass comprises a) providing a water-based emulsion of the vinyl polymer; b) mixing the water-based emulsion with at least one enzyme-mediator system to obtain a reaction mixture, said at least one enzyme-mediator system comprising at least one multi-copper oxidase chosen among a laccase, a laccase-ferroxidase, a laccase-like multi-copper oxidase or a mixture thereof, and at least one respective mediator, said at least one multi-copper oxidase and said at least one respective mediator being mixed sequentially with the water-based emulsion or provided as a pre-mix preserved under anaerobic conditions, until being mixed with the water-based emulsion; c) stirring the reaction mixture under an atmosphere comprising dioxygen; and d) recovering the at least one degradation product of the vinyl polymer.

The invention belongs to the technical field of protein engineering modification, and particularly relates to a method for improving thermal stability of amine oxidase ACAO and a mutant. Compared with a wild type enzyme, the temperature tolerance of the mutant enzyme in a certain temperature range is superior to that of the wild type enzyme, and the thermal stability is remarkably improved. Through molecular modification on a wild type, the thermal stability of amine oxidase ACAO mutant enzyme is remarkably improved. The enzyme is mainly applied to the industries of food production, livestock feed processing and the like so as to reduce the harm of fumonisins to the health of animals and human beings. The thermal stability improved amine oxidase ACAO mutant obtained by the invention is beneficial to improving the production efficiency of continuous production of enzyme in actual industrial application, has good industrial application prospects, and lays a foundation for further research on the structure of amine oxidase.

The invention discloses preparation and application of salt-tolerant amine oxidase KCYOBN sourced from bacillus subtilis and capable of degrading biogenic amine, and belongs to the technical field of molecular biology. The invention provides the amine oxidase from bacillus subtilis, and the expression of the amine oxidase in escherichia coli is realized. The invention also provides an application of the amine oxidase in degradation of biogenic amines, the amine oxidase is added into fermented food, the biogenic amines can be effectively degraded, an enzyme library for degrading the biogenic amines can be expanded, and the safety of the fermented food is further improved.

The invention discloses an application of a reagent containing multi-copper oxidase in reducing the biogenic amine content of a fermented meat product, recombinant multi-copper oxidase is obtained through a genetic engineering technology, and the multi-copper oxidase is developed into an enzyme reagent for degrading biogenic amine in Sichuan sausages. Results show that the addition of the reagent containing the multi-copper oxidase and the diamine oxidase has no influence on the eating quality and the sensory quality of the Sichuan sausages, in terms of the biogenic amine content, after the reagent is added, the degradation rates of histamine, tyramine, cadaverine and putrescine are at least 49.63%, 38.51%, 38.26% and 35.40%, and the degradation rates of the reagent containing the multi-copper oxidase and the diamine oxidase are at least 49.63%, 38.51%, 38.26% and 35.40%. The reagent provided by the invention has a good degradation effect on four main biogenic amines in Sichuan sausages.

Sensors, kits, and methods for assaying L-DOPA using engineered multi-copper oxidase are disclosed. Engineered multi-copper oxidase has improved catalytic activity at room temperature and can selectively dehydrogenate L-DOPA as a substrate when immobilized on an electrode for electrochemical detection. Embodiments include a sensor for assaying L-DOPA, the sensor comprising engineered multi-copper oxidase, which has increased catalytic activity toward the substrate compared to the catalytic activity of wild-type multi-copper oxidase.

The present application relates to a kind of collagen hydrogel and its preparation method and application, the preparation method includes the following steps: (1) using amine oxidase catalytic collagen oxidation deamination generates unsaturated aldehyde group functional group, intramolecular or intermolecular crosslinking occurs, completes first crosslinking;(2) the product of first crosslinking occurs secondary crosslinking under the catalysis of carboxyl activating agent, obtains the collagen hydrogel.In the present application, the collagen hydrogel prepared by the method is pure collagen hydrogel, solidification speed is fast, biocompatibility is good (excellent bionics), mechanical strength is adjustable, structure size is adjustable, stability is good, and application range is wide.

The invention discloses preparation and application of bacillus-sourced multi-copper oxidase LYSUF1 capable of degrading biogenic amine in fermented food, and belongs to the technical field of molecular biology. The invention provides the bacillus-sourced multi-copper oxidase, and the expression of the enzyme in escherichia coli is realized. The invention also provides application of the multi-copper oxidase in degradation of biogenic amines, the multi-copper oxidase is added into fermented food, so that the biogenic amines can be effectively degraded, an enzyme library for degrading the biogenic amines can be expanded, and the safety of the fermented food is further improved.

The invention discloses an application of a tobacco single copper oxidase-like protein gene NtSKU5, the nucleotide sequence of the tobacco single copper oxidase-like protein gene NtSKU5 is shown as SEQ ID NO.1, the tobacco single copper oxidase-like protein gene NtSKU5 comprises 1818 basic groups, and the gene is derived from tobacco. According to the invention, agronomic traits of an NtSKU5 gene edited plant and a control (untransformed) plant are investigated in a squaring stage and a mature stage under normal conditions, and the plant height, stem girth, waist leaf length, waist leaf width and leaf area of the NtSKU5 gene edited plant are all higher than those of the control (untransformed). In a word, the NtSKU5 gene is knocked out by utilizing a CRISPR / Cas9 mediated gene editing technology to obtain a gene editing plant which affects the growth and development of the plant, so that a genetic material and a theoretical basis are provided for the research on the single copper oxidase-like protein of the tobacco and the research on the growth and development of the tobacco.

Method for degrading a vinyl polymer A method for degrading a vinyl polymer having an initial molar mass in order to form at least one degradation product having a final molar mass lower than the initial molar mass comprises a) providing a water-based emulsion of the vinyl polymer; b) mixing the water-based emulsion with at least one enzyme-mediator system to obtain a reaction mixture, said at least one enzyme-mediator system comprising at least one multi-copper oxidase chosen among a laccase, a laccase-ferroxidase, a laccase-like multi-copper oxidase or a mixture thereof, and at least one respective mediator, said at least one multi-copper oxidase and said at least one respective mediator being mixed sequentially with the water-based emulsion or provided as a pre-mix preserved under anaerobic conditions, until being mixed with the water-based emulsion; c) stirring the reaction mixture under an atmosphere comprising dioxygen; and d) recovering the at least one degradation product of the vinyl polymer.

The invention belongs to the technical field of biosynthesis, and particularly relates to a method for catalytically synthesizing R-citronellal based on a double-enzyme system. According to the method for catalytically synthesizing the R-citronellal through the double-enzyme system, one-step series conversion from geraniol to the R-citronellal can be realized through the synergistic effect of the double-enzyme system of the copper oxidase and the old yellow enzyme, especially by matching with a proper coenzyme system and a metal salt system, so that the reaction efficiency and the enantioselectivity of a product are greatly improved, and the yield of the R-citronellal is increased. The method is a high-efficiency and high-selectivity synthesis system for synthesizing R-citronellal under the catalysis of a double-enzyme system.

The invention discloses laccase, cyclized laccase and application of the laccase and the cyclized laccase in detoxification of mycotoxin. According to the invention, a multi-copper oxidase laccase is screened from pediococcus pentosaceus, and in order to obtain a recombinant laccase with relatively good stability, a SpyTag / SpyCatcher technology is utilized to construct a mutant cyclized laccase. Through enzymatic property analysis of the two, the relative enzyme activity of the mutant cyclized laccase in temperature, pH, metal ions and organic solvents is higher than that of wild type laccase. According to the invention, the mutant cyclized laccase is adopted to reduce detoxification of aflatoxin B1 and zearalenone, and both the aflatoxin B1 and zearalenone show degradation activity. According to the present invention, the cyclic laccase is constructed, the degradation conditions of the cyclic laccase on the AFB1 and the ZEN are further optimized, the optimal degradation rates of the cyclic laccase on the AFB1 and the ZEN are respectively 100% and 98.97%, and in addition, the cyclic laccase has a certain toxin degradation effect on the AFB1 and the ZEN in the polluted feed sample.

This application relates to a collagen hydrogel, its preparation method, and use. The preparation method includes the steps of (1) catalyzing the oxidative deamination of collagen using amine oxidase to generate unsaturated aldehyde functional groups, thereby generating intramolecular or intermolecular crosslinks and completing primary crosslinking, and (2) subjecting the primary crosslinked product to secondary crosslinking under the catalysis of a carboxyl activator to obtain the collagen hydrogel. The collagen hydrogel prepared by this method is a pure collagen hydrogel that exhibits a fast setting rate, good biocompatibility (excellent biomimetics), tunable mechanical strength, tunable structural size, good stability, and a wide range of applications.

The invention discloses an amine oxidase mutant and application thereof in synthesis of chiral compounds, and relates to the field of biology. The amine oxidase mutant provided by the invention has mutation at any one or more sites of the 47 site, the 182 site, the 183 site, the 275 site, the 282 site and the 335 site of a wild type amino acid sequence, has relatively high catalytic activity on (S)-3-aminobutanol and relatively good stereoselectivity, can selectively oxidize (S)-3-aminobutanol under the condition that (R)-3-aminobutanol is not influenced, and has good application prospects. When the 4-hydroxy-2-butanone is applied to catalysis of (R)-3-aminobutanol feed liquid containing (S)-3-aminobutanol, the R-type ee value in the feed liquid can be effectively increased, additional refining and purifying steps are not needed, and the cost is remarkably reduced.

The present application relates to a collagen hydrogel, a preparation method therefor, and the use thereof. The preparation method comprises the following steps: (1) by using an amine oxidase, catalyzing collagen to undergo oxidative deamination to generate unsaturated aldehyde functional groups, and performing intramolecular or intermolecular crosslinking, so as to complete primary crosslinking; and (2) subjecting the primary cross-linked product to secondary crosslinking under the catalysis of a carboxyl activator so as to obtain the collagen hydrogel. In the present application, the collagen hydrogel prepared by the method is a pure collagen hydrogel, and has a high curing speed, good biocompatibility (excellent bionic properties), adjustable mechanical strength, adjustable structural size, good stability, and a wide range of application.

The invention discloses preparation and application of alcohol-resistant amine oxidase LYYOBN1 capable of degrading biogenic amine, and belongs to the technical field of molecular biology. The invention provides the amine oxidase from bacillus, and the expression of the amine oxidase in escherichia coli is realized. The prepared amine oxidase is used for degrading biogenic amine, the enzyme is added into fermented food, the biogenic amine can be effectively degraded in an environment containing ethanol or NaCl, an enzyme library for degrading the biogenic amine can be expanded, and the safety of the fermented food is further improved.