48 results about "Multienzyme complexes" patented technology

The invention relates to a preparation method of a hydrolyzed protein mixture and a protein peptide powder feed additive, wherein a multienzyme complex enzyme method is used for hydrolyzing vegetable protein to prepare the hydrolyzed protein mixture and protein peptide powder. The technology of the method is simple, and a non-starch polysaccharide enzyme and a proteinase are compounded so as to be capable of improving the hydrolyzation rate and degree of protein as well as the utilization rate of feed and reducing the production cost. Products prepared by the preparation method can be used as the feed additive to be capable of significantly improving the laying rate of egg-laying poultry, improving the weight increment of meat poultry, reducing the feed-meat rate and improving the production performance of table poultry as well as reducing the incidence rate of diarrhea in young animals and the feed-meat rate of piggy; and the preparation method is applied to aquatic livestock to be capable of improving the weight increment of fish and reducing the bait coefficient and content viscosity in the intestinal tract.

An expression system for producing a multi-enzyme complex, the system including a nucleic acid molecule containing a promoter operatively linked to a nucleotide sequence including multiple genes encoding multiple enzymes that are components of the multi-enzyme complex.

The invention provides a multienzyme complex preparation and a preparation method and application thereof. The multienzyme complex preparation is characterized by being prepared by adding water to multienzyme complex, wherein the weight ratio of the multienzyme complex to the water is 1:(50-100); and the multienzyme complex comprises the following components in parts by weight: 30-35 parts of laccase, 25-35 parts of xylanase, 10-20 parts of pectinase, 0.1-10 parts of amylase, 0.3-10 parts of lipase, 0.1-5 parts of tannase, 0.1-3 parts of coenzyme, 0.1-3 parts of stabilizing agent and 0.3-25 parts of diatomite. The multienzyme complex preparation can be applied to the preparation process of the fiber pulp for papermaking, the fiber pulp in accordance with the papermaking production requirement can be obtained without further adopting the high-temperature / pressure chemical cooking method in the pulping process, and the original lignin can be preserved to the greatest extent in the fiber pulp preparing process.

The invention provides a method for manufacturing a multienzyme-complex organic fertilizer by using Chinese herbal medicine. The technical proportioning scheme is that rice husk meal 30%, humic acid 30%, plant ash 19%, bone meal 10%, medical stone 3%, Chinese herbal medicine 3%, syrup 2%, saccharomycetes 1% and strain 2%. The manufacturing method comprises the following steps: (1) smashing raw materials and filtering the raw materials through a sieve with 80-100 meshes, (2) mixing filtered raw materials, the syrup 2% and the saccharomycetes 1% and adding water into mixture to perform fermentation, (3) performing pile turning for one time after fermenting for 24 hours, recovering temperature in a pile to normal temperature after 48 hours, adding the strain 2% into the pile according to the proportion and performing even mixing and package to produce organic fertilizer powder, and (4) pelletizing and packing the organic fertilizer powder according to the organic fertilizer pelletizing process, wherein organic fertilizer pellets are produce when the temperature in processing does not exceed 80 DEG C and pellet humidity is 20-30%.

The invention discloses a recombinant bacillus subtilis and a construction method and an application thereof, which belong to the technical field of genetic engineering. The method takes cell self functional membrane microdomain FMMs as a space frame, and constructs a multienzyme complex by using the specific marker proteins FloA and FloT, an artificial substrate channel is constructed, the metabolic burden of the cells can be effectively reduced, and the multienzyme complex is attached to plasmalemma , which facilitates the transport of a product from the intracellular to the extracellular. The recombinant bacillus subtilis constructed by the method efficiently synthesizes GlcNAc without affecting cell life activities, and a toxic intermediate metabolite GlcN-6-P can also be confined to the vicinity of the plasmalemma to reduce or eliminate the inhibition of cell viability. During a shake flask fermentation of a composite medium, the yield of acetylglucosamine in contrast bacterial strain BSGC is only 0.45 g / L, and the BSGAT acetylglucosamine output is increased to 5.29g / L, which is 11.76 times that of the contrast bacterial strain. The recombinant bacillus subtilis construction method of the invention is simple and convenient to use, and has the good application prospect.

The present invention is multifunctional health toothpaste and its production process. The multifunctional health toothpaste contains multienzyme complex, vitamins, lecithin and fluorine ion. The multienzyme complex comprises lipase, cellulase, pectinase, proteinase and amylase; and the vitamins include vitamin C, vitamin D and vitamin E. The multifunctional health toothpaste has high dental caries resisting effect, high detergency, and certain anti-inflammatory and sterilizing effect. In addition, the multifunctional health toothpaste has also functions of preventing tartar, resisting senility, raising resistance and strengthening immunity.

The invention relates to a tobacco processing active complex enzyme preparation for improving and enhancing tobacco quality and a preparation method thereof, relating to multienzyme complex preparation added before tobacco redrying. In the method, oxidation-reduction enzyme is mainly used to process precursor matter chromoplast having larger contribution rate on tobacco quality and protein degradation product preposing Maillard reaction. The proportion of the complex enzyme preparation is as follows: 12000 U / mL of glucose oxidase, 370 U / mL of chlorophyllase, 230 U / mL of carotinoid oxidase, 190 U / mL of nicotine dehydrogenase, PMSF inhibitor and EDTA complexing agent. The activity of protease in preparation condensed from a basic solution is equivalent to 36000 U / mL of neutral protease under the temperature of 40 DEG C. The preparation is diluted by 250 times to replace a secondary tobacco wetting agent, thus being capable of improving quality, promoting grade, stabilizing commodity shelf life, and winning warm praise from customers.

Disclosed are methods related to label-free biosensor cellular assays to classify multienzyme complex modulators in live cells. Disclosed are also methods related to the identification of purinosome disrupting Casein Kinase II (CK2) inhibitors, and methods related to the use of purinosome disrupting CK2 inhibitors as therapeutic agents for modulating CK2 activity and purine synthesis pathway, and for improving prevention and treatment of CK2 associated cancers, viral infection and inflammation conditions.

The invention discloses a process for extracting polysaccharide from milkvetch roots. The process is characterized by including the following steps of step a, using a dry milkvetch root powder as a raw material, placing the dry milkvetch root powder in a reaction vessel, and adding a mixed solvent to the reaction vessel; step b, heating a solution obtained after the mixing reaction in the step a, and adding a multienzyme complex to the mixed solution, wherein the multienzyme complex comprises cellulose and pectinase with the weight ratio of 1:1; step c, cooling the mixed solution obtained after enzymolysis reaction, and removing lower layer precipitates after the cooling to obtain supernatant; step d, concentrating the supernatant obtained from the step c to be paste, adding ethanol for precipitation, and performing filtering to obtain a milkvetch root polysaccharide crude product; and step e, refining the milkvetch root polysaccharide crude product to obtain a milkvetch root polysaccharide refined product. The process for extracting the polysaccharide from the milkvetch roots has the advantages of being simple in operation, conditions are easy to achieve, the yield rate of the polysaccharide is high, and the purity is high.

The invention relates to a TLL-CvFAP multienzyme complex as well as a construction method and application thereof, the construction method comprises the following steps: firstly constructing a TLL-linker-SpyCatcher recombinant protein engineering bacterium and a CvFAP-linker-SpyTag recombinant protein engineering bacterium, then performing expression to obtain a CvFAP-linker-SpyTag protein and a TLL-linker-SpyCatcher protein, mixing the two proteins outside a cell, and performing self-assembly to obtain the TLL-CvFAP multienzyme complex. The TLL-CvFAP multi-enzyme complex is used as a catalyst in the production of alkane (alkene) hydrocarbon from grease, and the catalytic efficiency of cascade reaction and the yield of alkane (alkene) hydrocarbon are improved.

The invention relates to construction and application of recombinant bacillus subtilis capable of synchronously secreting MTHase and MTSase. According to the construction and application disclosed bythe invention, self-assembly module plasmids pDGI-7S6-P43-PhoD-MTSase-spyCatcher-6xHis and plasmids pAX01-7S6-P43-PhoD-spyTag-linker-spyTag-MTHase-spyTag-6xHis are constructed; the constructed self-assembly module plasmids are transformed into B.subtilis WB800n thalli to obtain seamless integrative recombinant B.subtilis WB800n; and the recombinant strain is subjected to secretory expression and in vitro assembling of a MTSase-MTHase multienzyme complex body, and an experiment of dual-enzyme conversion for producing trehalose is performed, so that the conversion rate of the trehalose is increased to 81.5%.

The invention provides a recombinant bacillus subtilis, construction method and use thereof, wherein the cell's own FMMs are used as a space scaffold, and a multi-enzyme complex is constructed from specific marker proteins FloA and FloT, such that an artificial substrate channel is formed, and the cell metabolic burden is effectively reduced. The recombinant Bacillus subtilis of the invention can efficiently synthesize GlcNAc without affecting cell life activity, and can also limit the toxic intermediate metabolite GlcN-6-P near the plasma membrane to reduce or eliminate its inhibition on cell activity. In the process of shaking flask fermentation using complex medium, the yield of acetyl glucosamine of the control strain BSG-C was only 0.45 g.L−1, while that of BSG-AT, BSG-ATP, BSG-ATPB increased to 5.29 g.L−1, 6.22 g.L−1 and 8.48 g.L−1 respectively. The construction method of recombinant Bacillus subtilis is simple, easy to use and has a good application prospect.

The invention relates to a method for constructing a self-assembled expression double-enzyme strain and application. The method comprises the following steps of constructing MTSase-MTHase multi-enzymecomplexes with different proportions and different structures to obtain an escherichia coli recombinant. According to the invention, the MTSase-MTHase multienzyme complexes with different proportionsare discovered, and the obtained escherichia coli recombinant has a certain influence on the conversion rate of trehalose; through the MTSase-MTHase multienzyme complexs with the same proportion anddifferent structures, the influence of the obtained escherichia coli recombinant on the conversion rate of the trehalose is large; and a theoretical basis is provided for research on production of trehalose by fermentation of a multi-enzyme system.

In various embodiments a system is provided that displays heterologous proteins on the surface of a Gram-positive microorganism. In certain embodiments the system displays proteins using a sortase transpeptidase to covalently anchor proteins to the cell wall of the microbe. Novel bacterial strains are provided to exploit this system to display cellulase enzymes and multi-enzyme complexes on the surface of Gram-positive microorganisms (e.g., Bacillus subtilis) through their non-covalent interaction with a scaffoldin protein that is covalently anchored to the cell wall by the sortase transpeptidase. The surface displayed protein complexes contain enzymes capable of degrading cellulose into its component sugars at accelerated rates as compared to solutions of purified enzymes.

The invention discloses a kit for clinical detection of abnormal prothrombin. The kit comprises a magnetic particle PIVKA-II calibration product, magnetic particle suspension coated with PIVKA-II antibodies, magnetic particle PIVKA-II enzyme conjugate and sample diluent. The principle of a sandwich method is adopted to detect. Magnetic particles are coated with the PIVKA-II antibodies. Horseradishperoxidase labeled PIVKA-II antibodies are adopted to prepare the enzyme conjugate. An antibody-antigen-antibody multienzyme complex is formed through an immunological reaction. The complex catalyzesa luminescent substrate to emit photons, and the intensity of photoluminescence is proportional to the content of PIVKA-II. Operation is simple and convenient, sensitivity is high, specificity is high and the detection range is wide. The calibration product in the kit is liquid, and therefore detection errors due to the re-solubilization of calibration product are avoided, detecting results are more accurate, and clinical application is facilitated. The kit can be adopted in early diagnosis of hepatocellular carcinoma, monitoring of disease progression, evaluation of therapeutic effect and the like.

A cell may include heterologous polynucleotides encoding a multienzyme complex involved in the metabolic pathway of phenylpropanoids and biosynthesis of a vanilloid or a hydroxybenzaldehyde precursor thereof, which multienzyme complex comprises enzymes for the biosynthesis of coumaric acid and a crotonase.

The invention provides a multienzyme compound capable of preventing and treating fungus infection. The multienzyme compound of the invention can be used in daily prevention of fungus infection, and can kill fungi selectively without generation of toxic or side effects, or drug resistance. The multienzyme compound of the invention can treat fungus infection individually. Because the multienzyme compound can digest fungus cell walls, when cooperated with other antifungal medicaments to treat fungus infection, the multienzyme compound of the invention can promote penetration of other medicaments, enhance treatment effects of other medicaments effectively, reduce dosage and application amount, lower toxic and side effects, and reduce generation probability of drug resistance. In addition, the multienzyme compound of the invention can be selectively added with antifungal medicaments to improve treatment effects.

A polycistronic expression cassette encoding proteins necessary for constructing a multi-enzyme complex was developed. Also disclosed herein is a host cell containing this polycistronic expression cassette and uses thereof in degrading biomass.

A lignocellulolytic multi-enzyme complex in the form of a cellulosome, which includes a lignin-modifying enzyme and a carbohydrate-active enzyme, is provided herewith, as well as bifunctional chimeric enzymes having lignin and cellulose / hemicellulose degrading capacity. Also provided are methods of degrading lignocellulolytic biomass, and compositions and systems for effecting the same.

The invention discloses a method for producing winter jujube multienzyme complex probiotic through winter jujube pomace and belongs to winter jujube multienzyme complex probiotic production. The method includes 1, winter jujube pomace enzymatic hydrolysis; 2, centrifugal, concentration and sterilization, namely utilizing a centrifuge to separate mixed jujube paste after enzymatic hydrolysis, and obtaining original water jujube juice for standby and partial pomace; performing pressure reduction and concentration on the centrifuged original water jujube juice until the soluble solid content is of 20%, adding 0.2% of yeast extract, 1.9% of ammonium sulfate, 0.1% of zinc chloride, 0.1% of magnesium sulfate and 0.6% of potassium dihydrogen phosphate, and sterilizing; 3, inoculation and ventilation fermentation; 4, spray drying. By the aid of the method, resource is saved, the jujube pomace waste is converted into treasures for reuse, pollution and enterprise cost are reduced, load is reduced for enterprises, environment is protected, the advantage of 'eating and extracting winter jujube completely' is realized actually, and high promotion and application value is provided.

The invention provides recombinant escherichia coli for synthesizing glutathione and application of the recombinant escherichia coli. The N end of bifunctional glutathione synthetase GshF and the C end of polyphosphoric acid kinase PPK are connected with SpyCatcher and SpyTag respectively, two modified gene segments are integrated to the same plasmid vector at the same time, and the recombinant escherichia coli is obtained through conversion. After the obtained recombinant escherichia coli is subjected to high-density culture and optimization of the inducible expression condition, the maximum expression of a multi-enzyme compound is obtained, escherichia coli thalli are collected for efficient catalysis of whole cells or assembly of the multi-enzyme compound, centrifugation or membrane separation is directly conducted after the reaction is finished, and the cells or the multi-enzyme compound can be repeatedly used after being simply recycled. According to the process, the production cost of glutathione is effectively reduced, the utilization efficiency of a catalytic system is improved through efficient repeated utilization, the recombinant escherichia coli is simple to operate, and the application prospect is good.

The invention relates to a construction method of an in-vitro artificial scaffold protein mediated trehalose multienzyme complex. The construction method mainly comprises the following steps: constructing a self-assembled scaffold protein module recombinant bacterium WB800n-ScafCCR; constructing a recombinant bacterium WB800n-P43-phoD-treYC-cdoc of a self-assembled catalytic module; constructing arecombinant bacterium WB800n-P43-phoD-treZ-Ctdoc of a self-assembled catalytic module; constructing a recombinant bacterium WB800n-P43-phoD-cgt-Rfdoc of a self-assembled catalytic module; performingsecretory expression on the recombinant bacteria, performing self-assembly in vitro, and obtaining a recombinant trehalose multi-enzyme complex; the efficiency of preparing trehalose by catalyzing thetrehalose multi-enzyme complex is higher than the catalytic efficiency of mixed free enzymes; the cellulose microspheres can be used for producing high-quality trehalose after being immobilized.

The invention provides a zymogram analysis method for semi-cellulosome analogue and belongs to the technical field of industrial biology. The method disclosed by the invention is characterized in thatthe whole analysis process is composed of three parts: electrophoresis, zymogram analysis and proteinogram analysis. A continuous gradient non-modified gel without concentrated gel is adopted for electrophoresis; an off-on probe substrate is adopted for zymogram analysis; coomassie blue staining or silver staining is adopted for proteinogram analysis. According to the method, once electrophoresisis performed while zymogram and proteinogram information of a block of gel are collected; if multienzyme complex is generated in supernatant of microorganism fermentation liquor can be quickly and effectively judged; the zymogram analysis method is developed for screening microbial strains capable of generating cellulosome compounds and can be applied to the fields of industrial bioconversion, biological energy source, medical intermediate preparation, and the like.

The present invention relates to bio-engineered multi-enzyme complexes having synergistic enzyme activity comprising xylanases and optionally further comprising additional carbohydrate active enzymes. Such complexes are advantageous for degrading recalcitrant cellulosic biomass.

The invention relates to fresh kale slicing, enzyme-deactivation and green-maintaining processing equipment, comprising a slicing device, a conveying belt, an enzyme-linked ultrasonic decompressing processing device, a high-temperature high-pressure saturated vapor jet tunnel type thermostatic enzyme-deactivation device and a low-temperature high-pressure saturated vapor jet tunnel type thermostatic quenching device. The processing includes the steps of slicing fresh kale through the slicing device, sending the sliced kale in the enzyme-linked ultrasonic decompressing processing device throughthe conveying belt to perform green-taste removal, performing green-maintaining enzyme-deactivation in the high-temperature high-pressure saturated vapor jet tunnel type thermostatic enzyme-deactivation device and then performing green-maintaining browning-proof treatment through the low-temperature high-pressure saturated vapor jet tunnel type thermostatic quenching device. Since the sliced kaleis processed through a lipase activity composite inhibitor by the multienzyme complex combined ultrasonic decompressing technology, green-taste substance of kale can be removed and degraded effectively, lipoxidase activity can be deactivated, green taste of kale can be reduced greatly, and the problem of strong green taste of sliced fresh kale after processing is solved.

The invention relates to a preparation method of a hydrolyzed protein mixture and a protein peptide powder feed additive, wherein a multienzyme complex enzyme method is used for hydrolyzing vegetable protein to prepare the hydrolyzed protein mixture and protein peptide powder. The technology of the method is simple, and a non-starch polysaccharide enzyme and a proteinase are compounded so as to be capable of improving the hydrolyzation rate and degree of protein as well as the utilization rate of feed and reducing the production cost. Products prepared by the preparation method can be used asthe feed additive to be capable of significantly improving the laying rate of egg-laying poultry, improving the weight increment of meat poultry, reducing the feed-meat rate and improving the production performance of table poultry as well as reducing the incidence rate of diarrhea in young animals and the feed-meat rate of piggy; and the preparation method is applied to aquatic livestock to be capable of improving the weight increment of fish and reducing the bait coefficient and content viscosity in the intestinal tract.

This invention describes novel catalytically active cytosolic enzymes for triacylglycerol biosynthesis from eukaryotic systems. The complex from oleaginous yeast was enzymatically characterized, and was found to contain lysophosphatidic acid acyltransferase, phosphatidic acid phosphatase, diacylglycerol acyltransferase, acyl—acyl carrier protein synthetase, superoxide dismutase and acyl carrier protein. The triacylglycerol biosynthetic machinery rapidly incorporates free fatty acids as well as fatty acyl-coenzyme A into triacylglycerol and its biosynthetic intermediates. Lysophosphatidic acid acyltransferase, phosphatidic acid phosphatase and diacylglycerol acyltransferase from the complex were microsequenced. Acyl carrier protein, superoxide dismutase and diacylglycerol acyltransferase genes were cloned and expressed in bacterial system. The soluble triacylglycerol biosynthetic enzymes (lysophosphatidic acid acyltransferase, phosphatidic acid phosphatase, diacylglycerol acyltransferase) in yeast, rat adipocytes and human hepatocyte cell-line (HepG2) exist in the cytosol either as free enzymes or as a multienzyme complex.

This invention describes novel catalytically active cytosolic enzymes for triacylglycerol biosynthesis from eukaryotic systems. The complex from oleaginous yeast was enzymatically characterized, and was found to contain lysophosphatidic acid acyltransferase, phosphatidic acid phosphatase, diacylglycerol acyltransferase, acyl-acyl carrier protein synthetase, superoxide dismutase and acyl carrier protein. The triacylglycerol biosynthetic machinery rapidly incorporates free fatty acids as well as fatty acyl-coenzyme A into triacylglycerol and its biosynthetic intermediates. Lysophosphatidic acid acyltransferase, phosphatidic acid phosphatase and diacylglycerol acyltransferase from the complex were microsequenced. Acyl carrier protein, superoxide dismutase and diacylglycerol acyltransferase genes were cloned and expressed in bacterial system. The soluble triacylglycerol biosynthetic enzymes (lysophosphatidic acid acyltransferase, phosphatidic acid phosphatase, diacylglycerol acyltransferase) in yeast, rat adipocytes and human hepatocyte cell-line (HepG2) exist in the cytosol either as free enzymes or as a multienzyme complex.

An expression system for producing a multi-enzyme complex, the system including a nucleic acid molecule containing a promoter operatively linked to a nucleotide sequence including multiple genes encoding multiple enzymes that are components of the multi-enzyme complex.